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A “Game-Changer” – Groundbreaking Discovery Paves Way for ALS Cure

By University of Western Ontario May 29, 2024

Canadian researchers have discovered a potential pathway to curing ALS by targeting protein interactions, significantly supported by the Temerty Foundation’s philanthropy, with plans to advance to clinical trials in five years. Credit: SciTechDaily.com

A new potential treatment for ALS could advance to clinical trials within five years, supported by a $10-million donation from the Temerty Foundation.

In a groundbreaking Canadian discovery powered by philanthropy, a team of Western University researchers led by Dr. Michael Strong has uncovered a potential path toward a cure for amyotrophic lateral sclerosis (ALS).

The breakthrough, which illustrates how protein interactions can preserve or prevent the nerve cell death that is a hallmark of ALS, is the culmination of decades of Western research backed by the Temerty Foundation.

“As a doctor, it’s been so important for me to be able to sit down with a patient or their family and say to them, ‘We’re trying to stop this disease,’” said Strong, a clinician-scientist who has devoted his career to finding a cure for ALS. “It’s been 30 years of work to get here; 30 years of looking after families and patients and their loved ones, when all we had was hope. This gives us reason to believe we’ve discovered a path to treatment.”

ALS: Challenges and Breakthroughs

ALS, also known as Lou Gehrig’s disease, is a debilitating neurodegenerative condition that progressively impairs nerve cells responsible for muscle control, leading to muscle wastage, paralysis and, ultimately, death. The average life expectancy of an ALS patient post-diagnosis is a mere two to five years.

In a study published recently in the neurology journal Brain , Strong’s team found that targeting an interaction between two proteins present in ALS-impacted nerve cells can halt or reverse the disease’s progression. The team also identified a mechanism to make this possible.

“Importantly, this interaction could be key to unlocking a treatment not just for ALS but also for other related neurological conditions, like frontotemporal dementia,” said Strong, who holds the Arthur J. Hudson Chair in ALS Research at Western’s Schulich School of Medicine & Dentistry. “It is a game-changer.”

Dr. Michael Strong, Arthur J. Hudson Chair in ALS Research at Western’s Schulich School of Medicine & Dentistry, has discovered a protein that could lead to a treatment for ALS. Credit: Allan Lewis/Schulich School of Medicine & Dentistry

In virtually all ALS patients, a protein called TDP-43 is responsible for forming abnormal clumps within cells, which causes cell death. In recent years, Strong’s team discovered a second protein, called RGNEF, with functions that are opposite to TDP-43.

The team’s latest breakthrough identifies a specific fragment of that RGNEF protein, named NF242, that can mitigate the toxic effects of the ALS-causing protein. The researchers discovered that when the two proteins interact with each other, the toxicity of the ALS-causing protein is removed, significantly reducing damage to the nerve cell and preventing its death.

In fruit flies, the approach notably extended lifespan, improved motor functions, and protected nerve cells from degeneration. Similarly, in mouse models, the approach led to enhanced lifespan and mobility, along with a reduction in neuroinflammation markers.

The team’s path to discovery was paved by the Temerty family’s long-standing investment in ALS research at Western – support Strong calls “truly transformational.”

Pioneering Advances with Philanthropic Support

Now Strong and his team have set a goal to bring their potential treatment to human clinical trials in five years, a mission that is fueled by a new gift from the Temerty Foundation.

The foundation, established by James Temerty, founder of Northland Power Inc., and Louise Arcand Temerty, is investing $10 million over five years to power the next steps to bring this treatment to ALS patients.

“Finding an effective treatment for ALS would mean so much to people living with this terrible disease and to their loved ones,” said James Temerty. “Western is pushing the frontiers of ALS knowledge, and we are excited for the opportunity to contribute to the next phase of this groundbreaking research.”

The new gift by the Temerty Foundation brings the family’s total investment in neurodegenerative disease research at Western to $18 million.

“Dr. Strong’s relentless dedication to his field is matched only by the Temerty family’s deep desire to make a difference for the thousands of people around the world diagnosed with this devastating disease,” said Western President Alan Shepard. “The investment – and foresight – of the Temerty Foundation has accelerated progress in finding an effective treatment for ALS. We are grateful for the Temerty family’s commitment to life-changing research.”

“This is a pivotal moment in ALS research that could truly transform patient lives,” said Dr. John Yoo, dean at Schulich Medicine & Dentistry.

“With Dr. Strong’s leadership, our continued investment in the best tools and technology, and the visionary support of the Temerty Foundation, we are thrilled to be heralding in a new era of hope for patients with ALS.”

Reference: “Mitigation of TDP-43 toxic phenotype by an RGNEF fragment in amyotrophic lateral sclerosis models” by Cristian A Droppelmann, Danae Campos-Melo, Veronica Noches, Crystal McLellan, Robert Szabla, Taylor A Lyons, Hind Amzil, Benjamin Withers, Brianna Kaplanis, Kirti S Sonkar, Anne Simon, Emanuele Buratti, Murray Junop, Jamie M Kramer and Michael J Strong, 13 May 2024,  Brain . DOI: 10.1093/brain/awae078

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Scientists report that new gene therapy slows down amyotrophic lateral sclerosis disease progression

by Jakob Mjöbring, Umea University

There has been a breakthrough in the research on the disease amyotrophic lateral sclerosis (ALS). Scientists at Umeå University report that the disease progression in a patient with a particularly aggressive form of ALS disease has slowed down considerably with the use of a new gene therapy. After four years on the medication, the patient can still climb stairs, rise from his chair, eat and speak well, and live an active and socially fulfilling life.

"I consider this a breakthrough for the research we have conducted for more than 30 years, here at Umeå University and University Hospital of Northern Sweden. We have never before seen treatment results as effective as these, using any other treatment," says Peter Andersen, a neurologist and professor at the Department of Clinical Sciences at Umeå University.

"An important discovery is that it is now possible to considerably reduce the levels of the disease-causing SOD1 protein, and simultaneously measure a clear inhibitory effect on further disease progression . When we diagnosed the patient at the neurology ward in early spring 2020, the patient's prognosis was 1.5–2 years of survival at best. The patient has far, far exceeded expectation."

The patient is from a family in southern Sweden with a particularly aggressive form of ALS disease caused by a mutation in the SOD1 gene. When a relative was diagnosed with ALS, the patient left a blood sample for research purposes to the ALS research team at Umeå University but chose to not learn about the results of the genetic test.

However, the patient was a carrier of the disease gene, and after experiencing muscle weakness four years ago, the patient realized that he too was afflicted. The patient was immediately received by the medical team at University Hospital of Northern Sweden and was diagnosed with early stage ALS disease.

Since the summer of 2020, the patient has been a participant in the Phase III study evaluating a new gene therapy developed for patients with SOD1 mutations causing misfolding and aggregation of SOD1 protein in motorneurons. Every four weeks, the patient received the experimental treatment at a university hospital in Copenhagen in Denmark.

Biomarker reduced by almost 90%

At the time of diagnosis in 2020, the patient's levels of the substance neurofilament L—a biomarker indicating breakdown of nerve cells—was very high. Now, four years later, the levels are reduced by almost 90%.

"When the patient was diagnosed at University Hospital of Northern Sweden in April 2020, we measured the level of neurofilament L to be as high as 11,000 nanograms per liter, which is high even for an ALS patient. In the most recent sample, after 50 injections of the new drug, the level is down to 1,200 to 1,290, which is a substantial decrease of the disease indicator," says Peter Andersen.

"The normal level for a person in the patient's age group is below 560. In blood, the level of neurofilament has fallen back to normal levels, and was down to 12 during the latest hospital visit. The normal level is less than 13."

The patient's level of function, measured using the scale ALSFRSR, is reduced compared to a healthy individual (48 points) but has stayed at almost the same level, around 35 to 37 points, for the last 18 months—that means that the patient's functional level is reduced by approximately 26% compared to a healthy individual.

A person with this aggressive type of ALS gene mutation that the patient has typically loses 1–1.5 points every month. That means that without treatment, the expected disease progression would have been very fast and given rise to substantial disability within 6–12 months, and, most likely, have lead to the patient's death in 2021.

"That this patient, more or less unimpeded, still can climb stairs four years after disease onset, that is somewhat of a miracle to see," says Karin Forsberg, a neurologist and researcher at the Department of Clinical Sciences who works alongside Peter Andersen and has researched SOD1 and ALS for more than two decades.

"To have succeeded with a drug treatment in this way is a great success and an inspiration. But it does not in any way mean that the job is done. This is just the beginning. It is also important to remember that the drug in question does not constitute a curative treatment, but it seems able to put the brake on disease progression. It gives us great hope to further develop pharmaceutical treatments for ALS-patients."

There are many types of ALS disease, and only 2% to 6% has an ALS disease caused by a mutation in the SOD1 gene. Many have a familial form of the disease, but mutations in SOD1 have also been found in so-called sporadic cases of ALS.

"Whether this drug has a similar effect on other types of ALS disease is currently unknown. There is need for much more research on the subject," says Peter Andersen.

The patient can still do almost all things that he could do when he first joined the study in the summer of 2020—his speech is unaffected, and he manages to do everything himself, he mows the lawn, goes shopping, and takes care of his children. Mentally he also feels a lot better, mainly because he now dares to feel hope.

'This is only the beginning'

The study that the patient is participating in ends this summer. The medication is not yet available in Sweden, but it has been approved by the United States Food and Drug Administration, FDA, and on the 23 of February 2024 the European Medicines Agency, EMA, recommended the use of the drug on patients with SOD1 gene mutations within the European Union.

However, the New Therapies Council i Sweden has asked the regional health care providers not to prescribe the drug until a health economic evaluation has been provided by the Dental and Pharmaceutical Benefits Agency.

"Our next step is to study the results from the patients receiving this drug. It has worked for some, but not all have seen the same positive effect. It could be a question of dosage, or at which disease stage the treatment was initiated. Maybe additional drugs are required to completely stop the process? Those are questions we now have to try and answer. This is only the beginning," says Karin Forsberg.

She pictures a future where treatment will be given based on what type of ALS disease the patient has, and that it most likely will require a combination of drugs. She emphasizes that there is much research being conducted both in Sweden and internationally to find new drug targets so that equivalent drugs can be developed for patient groups with other types of ALS, and she is hopeful that it will come true.

"We can measure in samples collected from the patient that the disease process is ongoing, but the patient's body seems able to compensate. Even now, four years after the patient started taking this new gene therapy drug. The Swedish Ethical Review Authority approved participation in these studies and now, several years later, we, as well as ALS physicians in other participating countries, see a clear clinical effect on many treated patients," says Peter Andersen.

"The next step will be to get approval from the Swedish Ethical Review Authority to study the compensatory mechanisms that treatment with this drug seems to have activated. There might be an opportunity here to get insights into how previously unknown parts of the nervous system work, and to develop even better new drugs."

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Friday, June 21, 2024

Scientists identify genes linked to brain cell loss in ALS

NIH-funded study shines light on disease mechanisms, pointing to possible therapeutic targets

  In a small study, researchers have discovered how a set of genes could cause neurons to die in sporadic amyotrophic lateral sclerosis (ALS) . The results, published in Nature Aging , provide insight into the root causes of ALS and may lead to new ways to halt disease progression. The study was funded by the National Institutes of Health (NIH).

ALS is a progressive neurological disorder that attacks motor neurons, nerve cells in the brain and spinal cord that control muscles, leading to muscle weakness, paralysis, and eventually death. Most cases of ALS are sporadic and occur in people without a family history or other clear risk factors.

By analyzing the genetic profile of thousands of neurons from postmortem brain tissue from people who had ALS and from healthy donors, researchers identified higher levels of risk genes for ALS and frontotemporal dementia (FTD). The genes were especially prominent in Betz cells, a type of motor neuron, that express the marker THY1. In people with ALS, this was linked to disruptions in other neurons, hindering their ability to build, transport, and break down proteins. The genes—including SOD1 , KIF5A , and CHCHD10 —are among the most common associated with ALS/FTD.

Additional experiments showed that these changes may be connected to the toxic accumulation of the protein TDP-43, a defining feature of ALS and some cases of FTD. Therefore, higher levels of ALS risk genes in a distinct type of cell could trigger a harmful chain reaction that leads to widespread neuron loss.

Betz cell degeneration is a hallmark of ALS and is thought to occur early on when symptoms first appear. Understanding what makes these and other cells vulnerable to ALS could lead to new treatments that slow and even stop disease progression.

The team also explored how glial cells are affected by ALS. Glia are support cells that normally keep neurons healthy, but in ALS they can become dysfunctional and damage neurons, often accelerating their demise. Researchers analyzed genetic data from two kinds of glial cells and found genes related to cellular stress and inflammation. More research is needed to determine if glial cell dysfunction is a consequence or cause of neuron degeneration in ALS.

Together, the results enhance our understanding of why some neurons are more susceptible to ALS and identify potential novel therapeutic targets .

The study was supported by the National Institute of Neurological Disorders and Stroke (NINDS) (K08NS104270) and the National Institute on Aging (NIA) (P30AG062421). Single-cell sequencing and other resources were provided by the NIA-funded Massachusetts Alzheimer's Disease Research Center, one of 35 centers found across the United States.

  Amelie Gubitz, Ph.D., program director, NINDS, is available to discuss the study findings. To arrange an interview, please contact  [email protected] .

Limone, F., et al. “Single-nucleus sequencing reveals enriched expression of genetic risk factors in extratelencephalic neurons sensitive to degeneration in ALS.” Nature Aging . June 21, 2024. DOI: 10.1038/s43587-024-00640-0 .

NINDS is the nation’s leading funder of research on the brain and nervous system. The mission of NINDS is to seek fundamental knowledge about the brain and nervous system and to use that knowledge to reduce the burden of neurological disease.

About the National Institute on Aging (NIA) : NIA leads the U.S. federal government effort to conduct and support research on aging and the health and well-being of older people. Learn more about age-related cognitive change and neurodegenerative diseases via NIA’s  Alzheimer's and related Dementias Education and Referral (ADEAR) Center website . Visit the main NIA website for information about a range of aging topics, in  English and Spanish , and  stay connected .

About the National Institutes of Health (NIH): NIH, the nation's medical research agency, includes 27 Institutes and Centers and is a component of the U.S. Department of Health and Human Services. NIH is the primary federal agency conducting and supporting basic, clinical, and translational medical research, and is investigating the causes, treatments, and cures for both common and rare diseases. For more information about NIH and its programs, visit www.nih.gov .

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.cls-1{fill:#00a6d4;}.cls-2{fill:#fff;} discoveries magazine Discoveries

Regenerative medicine: a new path for als treatment.

Nov 17, 2023 Cassie Tomlin

A first-of-its-kind stem cell therapy for ALS passes a critical safety benchmark, advancing the search to slow down, reverse and prevent the disease. In a parallel study, investigators are growing patient-derived stem cells to model ALS, hoping to uncover its mechanisms and classify it with more specificity. Can the cure to this degenerative condition lie in the endlessly regenerative power of stem cells?

First, her hands stiffened. Then she developed a limp. Ashley Fisher was 48 when she was diagnosed with amyotrophic lateral sclerosis (ALS) . During the following three years, as the unstoppable disease took hold of her body, she lost the ability to work, hike by the beach or shop swap meets on Saturday mornings. So, she sought out what she could do with the time she had left: She enrolled in a clinical trial.

Even after paralysis stole her speech, Ashley consented to a five-hour spine operation, took immunosuppressive drugs for a year and underwent extensive testing at Cedars-Sinai ’s ALS Clinic , where a team monitored the impact of the procedure on her body—specifically, one of her legs. Their goal: to test the safety of a combination stem cell/gene therapy to treat the rare neurodegenerative disease, caused by the unexplained, unstoppable death of motor neurons in the brain and spinal cord.

Moments after Ashley’s death, in May 2021 at a hospital in Oregon, her daughter, Courtney Fisher Olsen, relayed to a nurse the urgent instructions impressed upon her in the previous months: Call Cedars-Sinai and ask them to collect Ashley’s spinal tissue.

“She made this research a priority, and she was really proud of it,” Courtney says. “She would do anything to be part of finding answers.”

Ashley, along with the 17 others in the study, gave investigators their only opportunity to make a critical advance: The trial proved, for the first time, the safety of the implantation into the lumbar spinal cord of specialized stem cells—neural progenitors—engineered to express a powerful growth factor known to protect neurons. The findings, published in September 2022 in Nature Medicine , cleared investigators to study the therapy’s efficacy and continue refining the approach they hope that, ultimately, will slow or stop the disease.

These patients are the heroes of this research. They knew we weren’t going to cure their disease, only pursue whether the cells and this surgical approach were safe. We are encouraged enough by this approach to proceed in an attempt to slow disease progression.”

Richard Lewis, MD

Dr. Richard Lewis is principal investigator of a stem cell trial at the ALS Clinic.

“These patients are the heroes of this research,” says  Richard Lewis, MD,  director of the Electromyography Lab and principal investigator of the study at the ALS Clinic. “They knew we weren’t going to cure their disease, only pursue whether the cells and this surgical approach were safe. We are encouraged enough with the results to proceed to more patients and attempt to slow disease progression.”

Until now, ALS has frustrated researchers with a notoriously impenetrable monolith. Only 5% to 10% of patients carry genes known to cause the disease. Without the ability to biopsy brain and spinal tissue, little is understood about its mechanisms. In the absence of biomarkers, physicians can only diagnose ALS after it has already taken hold, and the three Food and Drug Administration approved treatments do little to slow its progression. The highly specialized, resourceful clinicians at Cedars-Sinai ’s ALS Clinic, an ALS Association Certified Treatment Center of Excellence, can only leverage tools and technologies to support their chief goal: to preserve quality of life as patients become paralyzed and die.

Buoyed by breakthroughs in the study of stem cells, Cedars-Sinai investigators are challenging assumptions and evolving their questions about ALS. Because fresh progress in the disease is fueled by the body’s cells at their most naive state, the ALS Clinic team has embarked on a new clinical trial to test the safety of stem cell implantation directly into the cerebral cortices of ALS patients. Scientists are growing cells from ALS patients in petri dishes to model the disease. Having built the largest library of hyper-specific disease data, the team is reconsidering whether ALS is not one disease but a collection of conditions. They aim to differentiate between genetic and sporadic forms of ALS, and scour the models for the earliest signs of cellular decline. Every approach takes square aim at the ultimate questions: Why do patients develop ALS, and how can we stop the suffering?

A STARRING ROLE FOR STEM CELLS

For nearly 20 years, Clive Svendsen, PhD , executive director of the Board of Governors Regenerative Medicine Institute and the Kerry and Simone Vickar Family Foundation Distinguished Chair in Regenerative Medicine, has cultivated a multipronged sneak attack against neurogenerative disease. The approach aims to replace diseased astrocytes, star-shaped cells that support motor neurons. In ALS, diseased astrocytes play a role in motor neuron death, which causes paralysis. Glial cell line-derived neurotrophic factor (GDNF) is a potent growth factor that can protect motor neurons, but delivery to patients is difficult since GDNF is too large to cross the blood-brain barrier. So, years ago, Dr. Svendsen generated a line of human neural progenitors that can become astrocytes, and genetically engineered them to release GDNF.

In 2007, he and colleagues at the University of Wisconsin published a paper in PLOS One demonstrating that, when implanted into the lumbar spinal cords of rat models of ALS, the neural progenitors became astrocytes and released GDNF. After Dr. Svendsen joined Cedars-Sinai , further success showing the function and safety of the cells in animal studies earned his group an $8 million grant from the California Institute for Regenerative Medicine (CIRM). The team also was granted FDA approval, in 2016, to begin the 18-patient trial in which Ashley Fisher participated.

“Proving the safety and getting the cells to survive was a huge lift,” Dr. Svendsen says. “Drug development can stop at any point, so to have 18 patients and no safety issues—we’re very excited to move forward.”

IN HIS OWN WORDS: CLIVE SVENDSEN, PHD, ON THE PATIENT WHO INSPIRED HIS WORK

Clive Svendsen, PhD

“I had worked on Parkinson’s disease for years—but in 2002, in Wisconsin, I met Jeff Kaufman , a young patient with ALS. I was astounded at the horror of this disease—he had been an athletic man, a wonderful lawyer in his 30s with a lovely wife and four beautiful kids, when he was diagnosed. When I met him, he could only use his eyes to communicate through a computer. His first words to me were “Can stem cells help me?”

At the time, I was founder and co-director of the Stem Cell and Regenerative Medicine Center at the University of Wisconsin. After encountering Jeff, I dove into the literature and started asking questions. I realized that the approach we’d been studying in other neurodegenerative diseases—using astrocytes and GDNF as a protective strategy—could also protect motor neurons that die in ALS. So I switched my focus, applied for funding from the ALS Association and it’s all cumulated into the work we do now.

When I met Jeff, he had been fighting the disease for 10 years. He died at 54 in March 2010, and I’ve now been fighting for 20 years. While we don’t have the knockout blow yet, we have landed some punches, and I’m not going to stop.”

The Phase I/IIa study was overseen by Pablo Avalos, MD, associate director of Translational Medicine at the Board of Governors Regenerative Medicine Institute. A neurosurgical team, led by J. Patrick Johnson, MD, co-medical director of the Cedars-Sinai Spine Center and vice chair of the Department of Neurosurgery, injected the cells into the exact part of the spinal cord that controls movement in either the right or left leg. The procedure was found to be safe for all patients. Investigators also observed, as a secondary measure, that the cells slowed disease progression in the treated leg in some patients, though this did not reach overall statistical significance. ALS typically causes decline in both sides of the body at the same rate. Since patients received cells in only one side of the spinal cord, their own untreated leg acted as an “internal control” for comparison.

Postmortem spinal tissue revealed that the stem cells were still alive and producing GDNF in the treated side of the spine for up to three-and-a-half years after transplantation. Investigators continue to study postmortem spinal cord and brain tissue, searching for differences in dysfunction between lower motor neurons located in the spinal cord and upper motor neurons found in the motor cortex of the brain. While clinicians can detect ALS with electromyography and strength assessments that indicate lower motor neuron function, they have no measurable disease markers for upper motor neuron changes.

Frank Diaz, MD, PhD , a neuromuscular medicine specialist and assistant professor of Neurology, who diagnoses and treats patients in the ALS Clinic, is employing novel MRI techniques to identify markers of upper motor neuron dysfunction in postmortem brains and in living patients. He hopes to detect signal abnormalities in upper motor neurons. When paired with corresponding clinical data, this could support the development of more specific diagnostic tools, he says.

“Some patients do not have obvious clinical evidence of upper motor neuron dysfunction early on, which leads to delays in diagnosis and treatment, and even precludes their participation in clinical trials,” Dr. Diaz says. “Identifying markers of dysfunction will help us tremendously in understanding how the disease starts in the first place.”

Read: ALS and Genetics: What Do We Know?

MODELING DISEASE ORIGINS

In tandem with clinical trials, Cedars-Sinai investigators are leveraging patient-derived stem cells to model the disease in the lab and in animals.

Ritchie Ho, PhD , assistant professor of Neurology and Biomedical Sciences, who runs a lab at the Cedars-Sinai Board of Governors Regenerative Medicine Institute, is investigating motor neurons made from induced pluripotent stem cells (iPSCs) of ALS patients to identify the earliest signatures of disease and elucidate how disease pathways differ among patients based on their genes.

Dr. Ho’s research trajectory syncs with the groundbreaking discovery, in 2006, of iPSC technology. His PhD thesis focused on the mechanisms of stem cell reprogramming: the complex process of turning blood or skin cells back into a blank slate, so they can be coaxed, by the addition of specific protein growth factors, to become any type of cell in the body.

In a 2021 paper published in Cell Systems , Dr. Ho uncovered molecular differences between iPSC models of patients with sporadic and familial ALS. Though sporadic and familial disease lead to the same outcome, for the 90% of ALS patients without an identified gene associated with the disease, there is no explanation for its development.

“The hope is that, for those patients, we can use their own stem cells to analyze their genetic background,” Dr. Ho says. “Maybe the cause of their disease is genetic—there could be an undiscovered constellation of gene mutations. Maybe it’s not just one gene that causes ALS, but a series of unfortunate events, a critical mass gone wrong.”

This leads to questions about whether investigators can stratify ALS and its pathological processes to segment the disease more specifically based on whether the cells experience problems clearing proteins, processing energy or conducting other activities.

“ALS is potentially an umbrella disease—we don’t know, out of 1,000 patients, if there could be 100 different groups,” Dr. Ho says. “We’re coming to the point where the spheres of medicine and basic research are at a conjunction. We solve these unknowns through integrating patients’ genetics with how their cells behave in the petri dish and which RNA and proteins they express. Then we can connect these data patterns back to what’s going on in patients’ bodies as observed in the ALS Clinic and eventually develop truly personalized medicine. We hope to predict how a person will develop ALS and treat their cells before they degenerate.”

Read:  ALS Patient Matt Ashley Shares Learnings From 'Heartbreaking' Disease

BUILDING THE FUTURE OF ALS RESEARCH

At the Cedars-Sinai Biomanufacturing Center , Dr. Svendsen’s group, in collaboration with Dhruv Sareen, PhD, the center’s executive director, is building the largest-ever collection of iPSCs from ALS patients as part of the nationwide Answer ALS initiative. The stem cell lines are paired with an open-source repository of corresponding clinical, genetic, molecular and biochemical information, amounting to the most comprehensive collection of ALS data in history.

The project is groundbreaking in scale and specificity, and intended to encourage and enable researchers everywhere to leverage the data to develop deeper questions—and pursue answers. Earlier this year, Dr. Svendsen’s group and collaborators published a paper in Nature Neuroscience utilizing Answer ALS data to uncover disease subtypes.

Though stem cells have vastly expanded our insights into ALS, the grim reality of the disease and the lack of treatments are never far from mind for devoted investigators.

“This is how depressing it is—we started collecting the lines four years ago, and 700 of those 1,000 patients have died,” Dr. Svendsen says. “That’s why we’re determined to do these big projects to understand more. We’re just starting to crack the surface in uncovering clusters of patients with certain characteristics, and we’ve started breaking down questions about what causes it. You can’t design a drug treatment for a disease if you don’t know its cause.”

Challenges in finding a cause remain: No model of any disease is perfect. What might get lost or introduced into iPSCs during “reprogramming” to make them less reliable imitators of how the disease behaves in the body? Investigators hope to address this open question to improve the best tools we have.

In the Newsroom: Engineering the Next Generation of Cell and Gene Therapies

STEM CELLS CONTINUE TO SHINE

Last year, with an additional $12 million from CIRM, Cedars-Sinai investigators launched another first-ever, 16-patient safety trial, transplanting the GDNF-producing stem cells into the brain, in a region of the motor cortex that controls hand movement. The research team hopes the operation will leave patients with nothing worse than a scar under their hairline, and that they’ll see a positive effect on hand use, which the team will monitor in the ALS Clinic.

Clinical trials proceed slowly, and participant selection is a frustrating paradox: Patients must show early stages of paralysis to qualify, by which time they likely cannot regain long-term function of their bodies. But investigators hope that the process of proving safety will ultimately lead to an efficacy trial combining delivery to both the spinal cord and motor cortex.

“This is a protective strategy and it’s definitely about timing,” Dr. Svendsen says. “If all the motor neurons are dead, no matter how much progenitor cells and GDNF we deliver, they’ve got nothing to act on. In the future, we can intervene earlier in the disease, and that’s when we might start slowing the progression.”

Cell-based regenerative medicine isn’t the only approach that offers hope for ALS patients. Scientists at Houston Methodist are focused on immunotherapy: A 2022 study published in Neurology proved the safety of the intravenous application of T cells meant to slow ALS progression. Another trial, completed at six sites including Cedars-Sinai , met safety benchmarks for the infusion of ALS patients’ own bone marrow stem cells into their spinal fluid; but a Phase III study, published in a 2022 Muscle Nerve paper, did not meet its efficacy endpoint.

Dr. Lewis remains focused on stem cells’ potential for progress, and hopes that continued clinical trials will validate the approach.

“We are not going to be able to convert this into a therapy for people who currently have the disease,” he says. “But the scientific question is an important one, and the fact that the stem cells survive is crucial—it’s a start.”

TRICKING TIME IN A DISEASE OF AGING

Descriptions of major disorders like cancer, lung disease and heart disease date to at least 1500 B.C.—but ALS wasn’t identified until 1869, and wasn’t widely known until 1939, when baseball player Lou Gehrig was diagnosed.

Because ALS typically presents between the ages of 55 and 75, perhaps a shift in life expectancy—from under 40 in premodern times to a turn of the 20th century increase—could explain why the neurodegenerative disease was undocumented before the modern era. But if ALS is a consequence of our expanded longevity, what explains why it strikes in old age? What are the hazards in aging?

After discovering, as outlined in a 2016 Nature Neuroscience study, that motor neurons derived from iPSCs of ALS patients more closely resemble fetal cells than adult cells, Ritchie Ho, PhD, is attempting to “age” them in the lab to more faithfully model late-onset disease. He hopes to accelerate iPSC maturation—to synthesize a lifespan—and search for signs of pre-programmed cellular “events” that instigate the onset of ALS.

“If we can activate the aging program, we can start to see the neurons become diseased and die, and how,” Dr. Ho says. “We’re trying to look at the very early origins of ALS, assuming there’s something in the neuron that sets it up to die, and trying to understand what that is in order to intervene before it manifests in paralysis.”

To try to trick time, Dr. Ho is measuring gene expressions of iPSC-built motor neurons against gene expressions in tissue from ALS patients and healthy patients. He’s attempting to adjust the manufactured cells to better mimic adult cells by adding small molecules known to switch genes on and off.

In theory, the artificial, expedited aging of iPSCs can help identify what triggers disease onset, Dr. Ho says. “If we can see the full cycle of ALS from start to finish—how the cellular-energy expenditure is different, how proteins accumulate and, at the last stage, how cells are dying—we can pick it apart and see where along a lifespan the disease processes happen.”

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Breakthrough that can halt the progression of ALS developed by Northeastern scientist

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A major breakthrough in the treatment of amyotrophic lateral sclerosis, known as ALS, can potentially help stop the disease in its tracks in as much as half of the cases in the U.S., a Northeastern University scientist says.

Jeffrey Agar , associate professor of chemistry and pharmaceutical sciences at Northeastern, has spent the last 12 years studying the mechanism of ALS and researching ways to prevent its progression.

“You could consider it my life’s work,” he says. “I bet 12 years of my own life and countless number of years of others’ lives toward something that was so risky that everyone said it would never work.

“I am relieved that it all worked out.”

ALS is a rare progressive disease that causes deterioration of nerve cells in the brain and spinal cord. The disorder affects motor neurons, which control voluntary muscle movement, talking, walking, chewing and breathing. The onset of ALS is largely sporadic — only 10% to 20% of cases in the U.S. are inherited, Agar says, and therefore are called familial ALS (fALS). ALS can be caused by dozens of different gene mutations that lead to mutation in proteins within a cell.

In his research, Agar focused on the mutation of a protein called copper zinc superoxide dismutase 1 (SOD1), a major antioxidant. A mutation of SOD1 protein called A4V, he says, is one of the most common causes of familial ALS that often results in a patient’s death in less than a year.

The mutated protein splits into two toxic pieces called monomers, Agar says, that can stick to millions of other monomers. These monomers form toxic clusters in the cell that grow with the progression of the disease, damaging the cell and causing it to die. 

The novel treatment strategy developed by Agar’s lab uses a small molecule linker, S-XL6, to prevent the separation of the SOD1, stopping the mechanism that destroys cells. 

“Unlike Biogen’s approach, which diminishes SOD1 function, our method actually helps the protein regain its normal function,” Agar says. 

His experiments confirmed that this treatment method works in mice for a specific mutation of the SOD1 protein associated with familial ALS. In about 50% of all ALS cases there are no mutations in SOD1, but the protein is still being damaged trying to protect the cell from free radicals, Agar says, meaning that in the best-case scenario the therapy can potentially help halt the progression of the diseases and improve survival in half of the ALS cases in the U.S. 

“We’re making new molecules with t he Roman Manetsch Research Group , a medicinal chemistry lab at Northeastern, hoping to even improve it further,” he says.

Testing in mice, rats and dogs is promising, and the lab is moving forward with final testing for effectiveness and safety necessary for clinical trials. The treatment engages and stabilizes 90% of SOD1 protein in blood cells, Agar says, and 60% to 70% in brain cells at a safe dose.

“We made sure to never publish anything until we actually had an idea of the safety,” Agar says. 

Previous efforts of developing a compound that would stabilize SOD1 protein have not yet resulted in a treatment. Agar’s treatment won’t reverse the damage already done to the neurons and muscles, he says, because it is hard to reestablish vanished neuron connections.

This research was financially supported by the National Institute of Neurological Disorders and Stroke, part of the National Institutes of Health; the ALS Association; Johnston Educational Ventures; and the National Science Foundation.

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Agar also credits the success of his research to the longtime collaboration with Roman Manetsch , professor of chemistry and chemical biology at Northeastern, and all the doctoral students who worked in their labs over the years. 

“The thing that made all this possible was our growing industry Ph.D. program,” he says.

Under an agreement with Northeastern, employees of major pharmaceutical companies such as Novartis, Biogen or GSK come to the university to get a doctoral degree. They bring a variety of skills and knowledge about drug development, Agar says, that academics are not trained in.

In exchange, he says, the master’s level scientists from the industry are trained in analytical chemistry techniques — an exigent need for the pharmaceutical companies.

Agar’s lab is now developing a potential drug for ALS based upon this breakthrough.

“We want to get to clinical trials as fast as we can, only $4 million to go!” he says.

With this drug and other treatments currently available, such as Biogen’s Tofersen, which reduces the level of SOD1 protein in cerebrospinal fluid cells, Agar says, ALS patients potentially will be able to live a long life. 

“If you are diagnosed early, people might be able to still walk, talk and move on about life,” Agar says.

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ALS neuron damage reversed with new compound

• Feinberg School of Medicine

Northwestern University scientists have identified the first compound that eliminates the ongoing degeneration of upper motor neurons that become diseased and are a key contributor to ALS (amyotrophic lateral sclerosis), a swift and fatal neurodegenerative disease that paralyzes its victims.

In addition to ALS, upper motor neuron degeneration also results in other motor neuron diseases, such as hereditary spastic paraplegia (HSP) and primary lateral sclerosis (PLS).

In ALS, movement-initiating nerve cells in the brain (upper motor neurons) and muscle-controlling nerve cells in the spinal cord (lower motor neurons) die. The disease results in rapidly progressing paralysis and death.

So far, there has been no drug or treatment for the brain component of ALS, and no drug for HSP and PLS patients.

“Even though the upper motor neurons are responsible for the initiation and modulation of movement, and their degeneration is an early event in ALS, so far there has been no treatment option to improve their health,” said senior author Hande Ozdinler, associate professor of neurology at Northwestern University Feinberg School of Medicine. “We have identified the first compound that improves the health of upper motor neurons that become diseased.”

The study was published in Clinical and Translational Medicine on Feb. 23.

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Western University researchers unlock potential 'cure' for ALS

New research out of London, Ont.’s Western University is shedding light on a potential cure for ALS, in which the targeting of the interaction between two proteins can halt or fully reverse the disease’s progression.

According to a news release from Western University on Monday, a team of researchers, led by Dr. Michael Strong, have discovered a potential path toward a cure for amyotrophic lateral sclerosis, also known as ALS.

“As a doctor, it’s been so important for me to be able to sit down with a patient or their family and say to them, ‘We’re trying to stop this disease,’" said Strong, Arthur J Hudson Chair in ALS research at Western’s Schulich School of Medicine & Dentistry.

The discovery

Published in the journal Brain , Strong’s team discovered that targeting an interaction between two proteins present in ALS-impacted nerve cells can halt or even fully reverse the progression of the disease.

“Importantly, this interaction could be key to unlocking a treatment not just for ALS but also for other related neurological conditions, like frontotemporal dementia,” said Strong. “It is a game changer.”

According to the study, in nearly all ALS patients, a protein called TDP-43 is responsible for forming abnormal clumps within cells, which causes cell death. In recent years, Strong’s team discovered a second protein, called RGNEF, with functions that are opposite to TDP-43.

The team’s research identified a specific fragment of the RGNEF protein -- named NF242 -- that can “mitigate the toxic effects of the ALS-causing protein.”

As a result, the researchers discovered that when the two proteins interact with each other, the toxicity of the ALS-causing protein is removed, which significantly reduces damage to the nerve cell and therefore prevents its death.

After 30 years of his life working on ALS research with various teams, Strong is confident in what the future holds as a result of the new discovery not just for the treatment of ALS, but for other neurodegenerative diseases.

“I think we can reasonably look at this and say, ‘This is swinging for the bleachers,’ we think in three to five years, we might be looking at something [ALS] that might be cured,” said Strong. “We can, in an experimental model, significantly either completely abolish the [ALS] disease process. Or, in another one do a significant job of slowing it down and changing the course of the disease.”

How researchers got here

The light bulb moment in Strong’s research came seven years ago when the team was working with fruit flies. Flies that were genetically modified to have the pathology of ALS died within 10 to 14 days, whereas a fly typically has a lifecycle of 70 to 80 days.

Those flies with the new pathway Strong said had a longer lifespan, improved motor functions and their nerve cells were protected from degeneration.

The next step in research was using mice, where a mouse named Lucy who had been genetically modified to have ALS was given both treatments.

Her sister did not receive the treatment and died, whereas Lucy lived through experimentation and according to Strong was “doing fine.”

“Best results I’ve ever seen from an animal model,” he said.

What is ALS?

Also known as Lou Gehrig’s disease, ALS is a debilitating neurodegenerative condition that progressively impairs nerve cells responsible for muscle control, leading to muscle wastage, paralysis and death, according to Western.

The average life expectancy of an ALS patient post-diagnosis is two to five years.

Due to the complex nature of the disease, there are no effective forms of therapy that can prevent its progression.

A ‘gift’ for ALS research

The announcement was made ahead of ALS Awareness Month in Canada.

Also included in Monday’s announcement was a $10 million donation over the next five years from the Temerty Foundation.

Established by Jim Temerty, founder of Northland Power Inc., and Louise Arcand Temerty, the funding will help with the next steps of bringing the newfound treatment to ALS patients.

“Finding an effective treatment for ALS would mean so much to people living with this terrible disease and to their loved ones,” said James Temerty. “Western is pushing the frontiers of ALS knowledge, and we are excited for the opportunity to contribute to the next phase of this groundbreaking research.”

Strong and his team have set a goal to bring their potential treatment to human clinical trials within the next three to five years, “a mission that is fueled by the new gift from the Temerty Foundation,” Western said.

Strong is very cautionary, but if things progress he expects a “cure” is not far off.

“It’s been 30 years of work to get here; 30 years of looking after families and patients and their loved ones, when all we had was hope. This gives us reason to believe we’ve discovered a path to treatment," he said.

-- With files from CTV News London's Sean Irvine

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New treatment may help slow progression of ALS, research shows

An experimental medication may slow the progression of amyotrophic lateral sclerosis, or ALS, researchers reported Wednesday. The research was supported in part by donations from the Ice Bucket Challenge , the social media sensation that raised more than $200 million worldwide.

The drug is not a cure, but it may help slow the inexorable disability caused by ALS, which rapidly destroys the nerve cells that control the muscles that allow us to move, speak, eat and even breathe.

"Patients keep telling me their No. 1 goal is to be able to retain physical function for as long as possible," said the study's lead author, Dr. Sabrina Paganoni, a neuromuscular specialist at Massachusetts General Hospital's Sean M. Healey & AMG Center for ALS. "They want to be able to continue to walk and to use their hands."

About 20,000 people in the U.S. have ALS at any given time, according to the ALS Association. It usually strikes between the ages of 40 and 70. Once symptoms set in, life expectancy is two to six years, on average.

The treatment studied by Paganoni and her colleagues targets two cellular structures damaged by the disease: the mitochondria, which are the cells' power plants, and the endoplasmic reticulum, the cellular dump trucks that cart away waste that can gunk up the cells' machinery.

The multicenter, randomized, double-blind study is the second step — a phase 2 trial — in a three-step process required by the Food and Drug Administration for drug approval. In a double-blind study, neither the patients nor the researchers know who is receiving the drug. If a phase 2 study generates positive results, the FDA typically requires a larger and longer phase 3 trial.

To test the effectiveness of the two-drug combination, the researchers recruited 137 ALS patients who had become symptomatic within the previous 18 months. About two-thirds of the patients (89) received the drug, while the remaining third were given a placebo.

Participants were evaluated on a scale of 0 to 48, measuring the disabilities caused by the disease.

"By the time they entered the trial, on average, patients had already lost 12 points. Their baseline score was about 36, on average," Paganoni said. "Each question addresses a specific domain of function and is scored on a scale from zero to four."

For example, for walking:

3 = Early ambulation difficulties

2 = Walks with assistance

1 = Nonambulatory functional movement only

0 = No purposeful leg movement

During the six months of the study, patients taking the medication lost an average of 2.32 points less than those receiving placebos, a 25 percent better functional outcome.

"A 2- to 3-point change can mean the difference between being able to do an activity independently or with an assistance device," Paganoni said.

News An ALS patient's dilemma: End his own life peacefully, or die slowly of the disease?

The trial did not show a difference between medication and placebo in outcomes such as time to death, tracheostomy, or permanent intubation, or hospitalization. But that may be because it ran for just six months.

Paganoni suspects that, if it is approved, the new drug would be just one part of a cocktail of medications that would help to keep ALS at bay.

Because the trial showed that the medication might make a difference, all participants were offered the opportunity to stay on it or, in the cases of those who were given placebos, to start on it. The Mass General researchers will track how the patients taking the medication do in the long term.

Paganoni credited the Ice Bucket Challenge for getting her study and others going.

"The Ice Bucket Challenge was an important turning point in the fight against ALS," she said. "It put ALS on the map and raised awareness of the disease and attracted more investigators and investment to the research."

With the good news from the trial, the ALS Association hopes to persuade the FDA to allow other patients to have access to the drug, even before phase 3 trial results are available.

"It's very unusual for an ALS clinical trial to hit its primary endpoint, so we're very excited about it," said Neil Thakur, chief mission officer for the ALS Association. "It's the difference between being able to feed oneself versus being fed or needing versus not needing a wheelchair."

ALS experts cautioned against rushing ahead without more data.

"The current data are definitely positive, but they need to be replicated," said Dr. Martina Wiedau-Pazos, a neurologist who is director of the ALS Clinic and Research Center at UCLA. "This study has limitations, such as being small and lasting just six months. I think a phase 3 trial is needed, because, in the past, positive outcomes from phase 2 trials were not confirmed in phase 3 trials."

Another issue is that the study included subjects who had more rapid disease progression than normal, said Dr. David Lacomis, chief of the neuromuscular division at UPMC in Pittsburgh. "So it's unclear what the effects would be in the broader ALS population," Lacomis said via email.

While the new findings are promising, they are not "earth shattering," said Dr. Erik Pioro, director of the section of amyotrophic lateral sclerosis and related disorders at the Cleveland Clinic. "But it does add credence to the idea that other pathways are playing significant roles in ALS pathogenesis."

Results of the trial were published in the New England Journal of Medicine.

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Linda Carroll is a regular health contributor to NBC News. She is coauthor of "The Concussion Crisis: Anatomy of a Silent Epidemic" and "Out of the Clouds: The Unlikely Horseman and the Unwanted Colt Who Conquered the Sport of Kings." 

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• Open access
• Published: 21 June 2024

Single-nucleus sequencing reveals enriched expression of genetic risk factors in extratelencephalic neurons sensitive to degeneration in ALS

• Francesco Limone   ORCID: orcid.org/0000-0002-2482-8801 1 , 2 , 3   na1 ,
• Daniel A. Mordes   ORCID: orcid.org/0000-0002-8344-2140 1 , 2 , 4   na1 ,
• Alexander Couto 1 ,
• Brian J. Joseph   ORCID: orcid.org/0000-0001-9876-0021 1 ,
• Jana M. Mitchell 1 , 2 ,
• Martine Therrien   ORCID: orcid.org/0000-0003-3150-4052 2 , 5 ,
• Sulagna Dia Ghosh   ORCID: orcid.org/0000-0003-1833-7296 1 , 2 , 6 ,
• Daniel Meyer   ORCID: orcid.org/0000-0003-4720-9891 2 , 6 ,
• Yingying Zhang 1 , 2 ,
• Melissa Goldman 2 , 6 ,
• Laura Bortolin 2 , 6 ,
• Inma Cobos   ORCID: orcid.org/0000-0002-2043-1890 4 ,
• Beth Stevens 2 , 5 , 7 ,
• Steven A. McCarroll   ORCID: orcid.org/0000-0002-6954-8184 2 , 6 ,
• Irena Kadiu 8 ,
• Aaron Burberry 1 , 2 , 9 ,
• Olli Pietiläinen 1 , 2 , 10 &
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• Amyotrophic lateral sclerosis
• Motor cortex
• Neuroimmunology
• Oligodendrocyte

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by a progressive loss of motor function linked to degenerating extratelencephalic neurons/Betz cells (ETNs). The reasons why these neurons are selectively affected remain unclear. Here, to understand the unique molecular properties that may sensitize ETNs to ALS, we performed RNA sequencing of 79,169 single nuclei from cortices of patients and controls. In both patients and unaffected individuals, we found significantly higher expression of ALS risk genes in THY1 + ETNs, regardless of diagnosis. In patients, this was accompanied by the induction of genes involved in protein homeostasis and stress responses that were significantly induced in a wide collection of ETNs. Examination of oligodendroglial and microglial nuclei revealed patient-specific downregulation of myelinating genes in oligodendrocytes and upregulation of an endolysosomal reactive state in microglia. Our findings suggest that selective vulnerability of extratelencephalic neurons is partly connected to their intrinsic molecular properties sensitizing them to genetics and mechanisms of degeneration.

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Amyotrophic lateral sclerosis (ALS) is a neuromuscular disease with survival limited to 2–5 years from onset, the most common motor neuron disease in aging and the neurodegenerative disease with one of the earliest onsets, in the mid-to-late 50s 1 . Although specific genetic causes have been identified, most cases are sporadic (~90%), have no family history and unknown etiology 2 , thus rendering modeling of the disease difficult 3 . Variants in genes associated with ALS can contribute to a related disorder, frontotemporal dementia (FTD), leading to the view of ALS and FTD as clinical manifestations of shared molecular causes. Bulk RNA sequencing of ALS postmortem brains have identified differences 4 and similarities between sporadic and familial 5 cases and highlighted shared profiles 6 , 7 , 8 . While they provided valuable insights, these studies had limited resolution on the cell types altered by ALS.

The most striking feature in ALS–FTD are protein aggregates of TAR DNA/RNA-binding protein 43 (TDP-43) in over 95% of ALS cases and ~50% of FTD cases, mostly in neurons 9 , providing one shared mechanism. It is still unknown how familial mutations and sporadic onset might converge on the formation of these aggregates and how it specifically affects classes of extratelencephalic corticospinal motor neurons (CSMNs), that is, Betz 10 and von Economo cells 11 . Moreover, strong evidence demonstrated that cells other than neurons are key mediators of disease progression and it remains unclear how these might contribute to the disease 7 , 12 , 13 .

Methods to study heterogeneity at a single-cell level have rapidly advanced and their application to human postmortem brain tissue is beginning to emerge. In this Article, we applied single-nucleus RNA sequencing (snRNAseq) and in vitro human induced pluripotent stem cell modeling to investigate changes in cortical cell types in sporadic ALS (sALS). Our profiling identified the intrinsically higher expression of ALS–FTD risk factors in a specific class of extratelencephalic excitatory neurons. In patients with ALS, these neurons and other subclasses of ETNs selectively express higher levels of genes connected to unfolded protein responses and RNA metabolism. We found that excitatory neuronal vulnerability is accompanied by a decrease in myelination-related transcripts in oligodendroglial cells and an upregulation of a reactive, proinflammatory state in microglia. We provide a preliminary, insightful view of disruptions triggered in human motor cortices in ALS and implicate aging-associated mechanisms that link altered proteostasis and inflammation to specific cell types in ALS.

snRNAseq profiling of ALS cortex

We used snRNAseq to profile motor/premotor cortical gray matter from patients with sALS and age-matched controls with no neurological disease using Drop-seq technology 14 (Fig. 1a , Source Data Table 1 and Extended Data Fig. 1a–c ). After screening for RNA quality, 79,169 barcoded droplets from eight individuals were analyzed ( n  = 5 sALS and n  = 3 control), with a mean of 1,269 genes and 2,026 unique molecular identifiers (UMIs) (Extended Data Fig. 1d ). We used Seurat 15 , the single-cell analysis package, to cluster and annotate groups according to canonical markers of brain cell types 16 : excitatory and inhibitory neurons, oligodendrocytes, oligodendrocyte progenitor cells (OPCs), microglia, astrocytes and endothelial cells (Extended Data Fig. 1e,f ). The observed cell type distribution corresponded to previous studies 16 and enabled robust categorization for downstream analysis. Cellular distribution was homogeneous between sexes and individuals, except for a modestly lower number of astrocytes in ALS samples (Extended Data Fig. 1g–i ).

a , Diagram of the workflow for isolation of nuclei from cortices of patients with ALS and age-matched controls followed by snRNAseq and assessment of expression of gene modules associated with neurodegenerative diseases. Exc, excitatory neurons; Inh, inhibitory neurons; Oligo, oligodendrocytes; OPCs, oligodendrocyte progenitor cells; Micro, microglia; Astro, astrocytes; Endo, endothelial cells. b – d , t SNE projections and violin plots of z -scores for expression of genes associated with ALS–FTD ( b ), AD ( c ) and MS ( d ) in the different cell types identified (the bars denote median for each side of the violin and the symbols indicate the average score per individual). e – g , t SNE projections and violin plots of z -scores for expression of genes associated with the ALS–FTD ( e ), AD ( f ) and MS ( g ) in the different subtypes of excitatory neurons (the bars denote median for each side of the violin and the symbols indicate the average score per individual) (in a – g , n  = 3 control individuals and n  = 5 patients with sALS).

Elevated expression of ALS–FTD genes in a specific class of neurons

To potentially identify cell types underlying ALS pathophysiology, we examined the expression of known familial genes for ALS–FTD and variants identified as risk factors from genome-wide association studies (GWAS). These genes were expressed to a variable degree between cell types and many of them were ubiquitously expressed as already known 2 , 7 (Extended Data Fig. 2a ). We then computed a ‘module score’ for this set of genes 17 ; this metric generates a standardized z -score for the expression of each gene and sums it up as a total score for the gene set. A positive score suggests higher expression of the gene set compared with the average expression of the module across the dataset. We computed parallel module scores for lists from the latest GWAS for disorders that affect the cortex but not specifically Betz cells: Alzheimer’s disease (AD) 18 , 19 and multiple sclerosis (MS) 20 (Fig. 1a and Source Data Table 2 ). No clear enrichment for the ALS–FTD gene list was identified (Fig. 1b ), as anticipated by the scattered, ubiquitous expression pattern. On the other hand, AD and MS modules showed respective enrichment in microglia, as expected on the basis of the strong immune signature in these diseases 18 , 19 , 20 (Fig. 1c,d ). These results corroborate knowledge in the field, underlying the strength of this analysis, and confirm our results in an unbiased, single-cell resolution.

Considering the selective loss of neurons in ALS 2 , we further analyzed these cells. We found 32,810 nuclei from excitatory neurons with unbiased clustering identifying seven subgroups (Exc0–6) expressing known markers of different cortical layers 16 (Extended Data Fig. 2b–f ). Analysis of the ALS–FTD module in these cells showed a positive score in THY1 -expressing subgroup Exc1 (normalized enrichment score of 1.834) (Fig. 1e and Extended Data Fig. 2g,h ) and no significant enrichment for AD and MS modules (Fig. 1f,g ). We decided to investigate the identity of these cells and the possibility of them being ETNs.

We identified three subgroups expressing markers of subcerebral projection neurons: Exc1, Exc5 and Exc6 (Fig. 2a ). Exc5 and Exc6 expressed canonical genes FEZF2 , BCL11B and CRYM 21 ; Exc1 expressed THY1 , enriched in human layer V 22 and used as a reporter for CSMNs 23 , and high levels of neurofilament chains, markers of ETNs 24 (Fig. 2b ). Recent reports dissected the transcriptomic identity of layer V extratelencephalic neurons in the human motor cortex 24 . We detected ETNs markers in these groups, with Exc1 expressing SERPINE2 and POU3F1 , specific of ETNs 24 , and NEFH and STMN2 , broad markers of motor neurons (MNs) 25 (Fig. 2c ). Owing to the anatomical location of our samples and the presence of ETNs across motor-related areas, we plotted genes specific to layer V ETNs von Economo cells 26 , 27 , affected in FTD and other long-range subcerebral projecting neurons (LR-SCPNs) 28 and confirmed that all three groups expressed these markers (Fig. 2d,e ). To further characterize these patterns, we leveraged a publicly available single-cell, spatial dataset of control human dorsal cortex 29 . We confirmed that markers expressed in Exc1 THY1 , STMN2 and SNCG (Fig. 2f ) are specifically expressed in layer V (L5) (Fig. 2g,h and Extended Data Fig. 3a,b ). This evidence suggests that Exc1, Exc5 and Exc6 express markers of layer V extratelencephalic neurons of cortical areas affected by ALS–FTD.

a , t SNE projection of presumptive layer V neurons. b , A dot plot representing expression of layer V markers c , A dot plot for markers of LVb ETNs of human motor cortex. d , A dot plot representing expression of von Economo markers. e , A dot plot representing expression LR-SCPN markers. f , Representative violin plot for markers of layer V ETNs of human motor cortex (geometric box plots for median and interquantile ranges). g , Visual depiction of layers identified by Maynard et al. 2021 ( n  = 7, g – j , publicly available) (L, layer; WM, white matter). h , A spot plot depicting expression of layer Vb motor cortex marker, STMN2 , identified as enriched in THY1-Exc1, with corresponding box plot quantification. i , j , Box plots ( j ) and corresponding spot plots ( i ) for the expression of the top five ALS–FTD-associated genes expressed in Exc1 (box plots for mean and interquantile ranges).

Source data

To further confirm that THY1 high neurons expressed higher levels of ALS–FTD genes, we ran a module score analysis in two datasets that identified THY1 high excitatory neurons 22 , 30 . In these studies, THY1 neurons expressed ETN markers, layer V, von Economo and LR-SCPN markers (Extended Data Fig. 3c–i ) and expressed higher levels of the ALS–FTD module (Extended Data Fig. 3l–m ). Analysis of the spatial transcriptomic dataset 29 confirmed that the top ten ALS–FTD-associated genes most highly expressed in Exc1 (Extended Data Fig. 2g ) are expressed in deeper layers of the cortex and specifically in layer V (Fig. 2i,j and Extended Data Fig. 3n ). Larger cohorts of patients and validations at the protein level are needed to confirm the degree of dependency of ETNs on this gene set but studies in human 31 and mouse 32 showed that deep-layer neurons have a higher propensity to form TDP-43 aggregates, the hallmark of ALS–FTD. Here, we provide a possible link to their specific vulnerability.

Higher cellular burden in deeper-layer excitatory neurons

We next examined whether the enriched expression of ALS–FTD genes might relate to changes that occur in excitatory neurons in response to ALS. We conducted differential gene expression (DGE) analysis between neurons from patients and controls, across all excitatory cells and within each subgroup (Fig. 3a ). We selected genes significantly upregulated in patients globally (DGEall) and within each subgroup (DGE0–6), calculated module scores for each set and investigated whether certain neuronal subtypes might have similar responses to ALS (Source Data Table 3 ). We found a correlation between scores in groups expressing deep-layer markers and the global changes identified in patients (Fig. 3b ), suggesting that pathology in lower cortical layers might be driving observed alterations. For instance, groups expressing ETN markers (Exc1, Exc5 and Exc6) shared many upregulated genes with each other and with the global signature (Fig. 3c ). Intriguingly, this class of genes is, like genetic risk factors, constitutively expressed at higher levels in Exc1 ETNs (Fig. 3b ), advocating for a proposed interplay between genetics and molecular pathways that sensitizes ETNs to ALS 33 .

a , A schematic of DGE analysis. b , A dot plot representing scores for genes upregulated in each subgroup of Exc neurons (DGE0–6) and globally upregulated in all Exc (DGEall). c , Comparison of genes globally upregulated in ALS (DGEall) with genes upregulated in classes of L5-ETNs (genes expressed by >10% of cells, >2 FC and adjusted P value <0.05). d , Violin plots of z -scores for genes globally upregulated in all excitatory cells (DGEall) in all excitatory neurons ( n  = 3 controls, n  = 5 patients with sALS; geometric box plots represent median and interquantile ranges and symbols indicate average score per individual). e , Violin plots of z -scores for genes upregulated in classes of L5 ETNs (DGE1, DGE5 and DGE6) in the three groups (geometric box plots represent median and interquantile ranges, symbols indicate average score per individual). f , GO analysis for genes upregulated in L5 ETNs classes (DGE1, DGE5 and DGE6); the highlighted terms are shared between the three. PD, Parkinson’s Disease; AD, Alzheimer’s Disease; HD, Huntington’s disease; CC, cellular components; KEGG, Kyoto Encyclopedia of Genes and Genomes. g – h , Western blot quantification of ubiquitin accumulation and 20S proteasome (prot.) subunit from motor cortices of separate cohort of patients with ALS ( n  = 6) and age-matched controls ( n  = 7) ( t -test).

Owing to the small cohort size and because of the diverse etiology of ALS–FTD 6 , we decided to test how shared these changes were and test whether they were driven by a single individual. We started by averaging gene expression by sample within groups of ETNs and ran principal component analysis (PCA) on the individual-aggregated matrices that showed widespread diversity between both patients and controls with partial segregation by diagnosis (Extended Data Fig. 4a ). This could be explained by a compound effect of diversity of human population, sampling of different cells per individual or different disease etiology. To avoid differential gene expression analyses confounded by this effect, we carried out several preanalyses to corroborate the strength of our study and come to more confident conclusions.

We proceeded to compute module scores for the global sALS signature (DGEall) and sALS signatures in ETNs (DGE1,5,6) and saw that, even though with intragroup variability, these signatures showed higher scores in ALS at single-cell and aggregated expression level with similarities between groups of ETNs (Fig. 3d and Extended Data Fig. 4b ). To ensure that these signatures were driven by disease changes, we recalculated differentially expressed genes (DEGs) by down-sampling each group of ETNs to the smallest number of cells by diagnosis (that is, redoDGE1,5,6). This analysis confirmed the great overlap of DGE signatures and redoDGE signatures and that these signatures were higher in sALS at single-cell resolution and average level (Extended Data Fig. 4c,d ).

Finally, to further test whether the results were driven by the transcriptome of a single individual, we repeated the DGE analysis by excluding cells from one patient at a time. This analysis shows that, even though different subsets of genes are detected in each analysis, the shared ones show a similar direction and amplitude in changes (Extended Data Fig. 4e ). This comparison shows that singular gene changes might be sporadically driven by one individual and because of the small cohort size, it would be hard to discern between biological or technical outliers. However, the shared DEGs (>65% in this case; Extended Data Fig. 4f ) and the general direction of the ALS-driven signature are maintained, with genes commonly unregulated in at least four out of five patients (ALLminus1 list) having >85% overlap with DGEall (Extended Data Fig. 4g ). We proceeded to use only genes shared by at least four individuals for subsequent analyses for all cell types.

We ran Gene Ontology (GO) analysis and showed that DEGs identified in classes of ETNs are connected to cellular stresses previously associated with ALS even from studies with hundreds of patients 7 , 34 (Fig. 3f ). Interactome analysis revealed coordinated alterations in genes that function in translational machinery, mitochondria, protein folding, proteostasis and degradation pathways connected to the proteasome and shared many transcriptional changes with patients’ excitatory cells as a whole (Extended Data Figs. 4k–m and 5 ). Interestingly, genes upregulated in upper layers of the cortex, a region relatively spared of pathology, shared less similarities with DGEall and were associated with synaptic biology (Extended Data Fig. 4h–j ). Comparison of ALS-driven transcriptomic changes with other studies underlined similarities with genes upregulated in excitatory neurons from MS 22 but not from AD patients 35 (Extended Data Fig. 4n,o ), suggesting that there are similar processes at the origin of neurodegeneration but that these changes are not universal. The analyses so far have highlighted the interindividual variability intrinsic of this kind of datasets and the particular attention studies need to divert into assuring reproducibility of the results. Nonetheless, we provide a snapshot of disruption in neuronal health in patients with ALS, in which lower layers of cortical excitatory neurons share higher levels of cellular stresses.

Next, we sought to determine what proportion of this transcriptomic signature may be associated with proteostatic stress specifically in neuronal cells. Presently, in vitro modeling of sporadic ALS requires high numbers of lines, high-throughput methods and needs further standardization 3 , 36 . To overcome these limitations, we implemented transient proteasome inhibition as a highly reproducible, dose-responsive, temporally controlled model to induce TDP-43 nuclear loss as seen in patients’ Betz cells and other ALS-related dysfunction in human neurons 4 , 37 , 38 (Fig. 4a and Extended Data Fig. 6a,b ). Application of a proteasome inhibitor to human pluripotent stem (hPS) cell-derived neurons 37 induced nuclear loss of TDP-43 (Fig. 4b ). Bulk RNA sequencing analysis showed widespread changes after treatment, with a significant overlap of upregulated genes between stressed hPS cell-derived neurons and sALS neurons (Fig. 4c ), specifically proteasome subunits and heat-shock response-associated chaperonins, confirmed by GO analysis of shared genes (Fig. 4d and Extended Data Fig. 6d ). Moreover, genes upregulated after inhibition show a significant overlap with transcripts misregulated after downregulation of TDP-43 in neurons 37 (Extended Data Fig. 6e ). This confirms that some changes identified in sALS neurons are connected to neuronally intrinsic proteostasis and are at least in part connected to alterations in TDP-43.

a , Diagram of neuronal differentiation from PSCs and treatment with proteasome inhibitors for bulk RNA sequencing. b , Immunofluorescence of TDP-43 localization after treatment. c , Venn diagram depicting shared upregulated genes between treated hPS cell-derived neurons and excitatory neurons from patients with ALS. d . GO analysis for shared genes in c , highlighted terms involved in protein folding and neurodegenerative diseases. CC, cellular components.

To confirm hindered proteostasis in ALS cortex, we selected a second cohort of patients with sALS and controls for biochemical evaluation. We extracted protein, confirmed increased insoluble TDP-43 in patients (Extended Data Fig. 6f,g ) and showed that, despite the presence of core proteosomal subunits, pathology is accompanied by the accumulation of highly ubiquitinated proteins (Fig. 3g,h ), the hallmark of impaired proteostasis. These findings suggest that the neuronal stress observed in ETNs from patients with ALS represents an intrinsic susceptibility to proteostatic stress orchestrated, in part, by abnormal homeostasis of TDP-43.

Oligodendroglia respond with a neuronally engaged state

To extend deep into the cord, ETNs are dependent on robust axonal integrity 39 . As others detected changes in myelination in ALS 12 and in FTD 40 , we analyzed 19,151 nuclei from myelinating cells that clustered into five groups: one of OPCs—oliglia3, and four of oligodendrocytes—oliglia0, 1, 2, 4 (Fig. 5a–c and Extended Data Fig. 7a,b ). We noted a significant shift of ALS nuclei representation in oliglia0 versus oliglia1 and oliglia4 (Fig. 5d ). Control-enriched oliglia0 were characterized by GO terms connected to oligodendrocyte development and myelination and expressed higher levels of myelinating genes, for example, CNP , OPALIN and MAG (Fig. 5e and Extended Data Fig. 7c,d ). ALS-enriched oliglia1 showed terms for neurite morphogenesis, synaptic organization and higher expression of synaptic-related genes DLG1 , DLG2 and GRID2 (Fig. 5f and Extended Data Fig. 7e,f ). Intriguingly, expression of neuronal enriched transcripts has been found in oligodendrocytes in primate motor cortex 24 .

a , t SNE projection of OPCs and oligodendrocytes markers. b , t SNE projection of oligodendroglia (ALS n  = 8,372 nuclei and control n  = 11,168 nuclei). c , t SNE projection of subclusters within oligodendroglia (Wilcoxon–Mann–Whitney). d , Distribution of subclusters by diagnosis) (mean ± s.e.m. e , GO analysis for genes characteristic of control-enriched oliglia0 highlighted terms involved in myelination. CC, cellular components. f , GO analysis for genes characteristic of ALS-enriched oliglia1 highlighted terms involved in neuro-engaged functions. g , Violin plots of representative genes for neuro-supportive functions (left) and myelination (right) (geometric box plots for median and interquantile ranges; symbols indicate log 2 (average expression) per individual (fraction of cell expressing). h , Volcano plot of DEGs in oligodendroglia. Highlighted genes identified in GO terms related to myelination (orange) and neuro-engaged functions (green). i , Violin plots representing z -score for selected GO terms and related t SNE projection (boxplot representing median and interquantile ranges; symbols indicate average score per individual). j , k , Western blot ( j ) and quantification ( k ) of CNPase and MBP from motor cortices of patients with ALS and age-matched controls ( t -test). l . Diagram illustrates shift of oligodendrocytes states ( t -test) (for a – l , n  = 3 control and n  = 5 patients with sALS). nUMI - normalized Unique Molecular Identifier.

While intragroup variability was apparent between aggregated oligodendroglial transcriptomic signatures, there was a demarcated separation of individuals by diagnosis with global gene expression conserved at single-cell and aggregated levels (Extended Data Fig. 7g,h ). We proceeded to calculate DEGs by excluding one individual at a time (ALLminus1 approach described above), selected genes commonly unregulated in at least four out of five patients and compiled gene lists for subsequent analyses (Extended Data Fig. 7i–k ). Altogether, DGE and GO analyses support a shift of sALS oligodendrocytes from myelinating to neuronally engaged states with upregulation of genes involved in synapse modulation and decrease of regulators of myelination (Fig. 5g–i and Extended Data Fig. 7l–o ). Loss of myelination is exemplified by the changes in G-protein coupled receptors (GPRCs) marking developmental milestones: GPR56 , expressed in OPCs 41 , and GPR37 , expressed in myelinating cells 42 , were lowly expressed in ALS-enriched groups and globally downregulated (Extended Data Fig. 7p ).

To further explore these changes, we compared them to published reports that identified shifts in oligodendrocytes in MS (Source Data Table 4 ) 43 . Comparison of Jäkel et al. 43 with our study revealed that control-related oliglia0 most closely resembled highly myelinating OPALIN + cells from Jäkel6 (Extended Data Fig. 8a–c ), while ALS-associated oliglia1 and oliglia4 aligned to not-actively myelinating Jäkel1 (Extended Data Fig. 8d–h ). To confirm this shift, we ran validations on protein extracts from motor cortices and showed that oligodendrocyte-specific, myelin-associated proteins CNP and MBP are downregulated in patients (Fig. 5j–k ), consistent with studies identifying demyelination in patients with sALS 12 and with bulk RNA sequencing studies that identified a decrease in myelinating markers 7 . The data so far show how activation of stress pathways in deep-layer neurons is juxtaposed to a shift in oligodendrocytes from active myelination to oligo-to-neuron contact (Fig. 5l ).

Microglia activate an endolysosomal response

Mouse models 44 , patient samples 5 and function of ALS-related genes in myeloid cells 45 have demonstrated the importance of microglia as modifiers of disease. In the 1,452 nuclei from microglia (Fig. 6a ), comparative DGE analyses showed intragroup variability in magnitude but conserved directionality of changes and robustness of the core common biology (Extended Data Fig. 9a–e ). We identified 159 genes upregulated in patients and, remarkably, many were associated with endocytosis and exocytosis, previously implicated in ALS 45 (Fig. 6b,c ). Several of these genes were also associated with microglial activation and neurodegenerative disorders ( CTSD, SPP1, CPM and APOE ) (Fig. 6c,d ) and interestingly with ALS–FTD ( TREM2 , OPTN , SQSTM1/p62 and GRN ) (Fig. 5e,f ). GO analysis for upregulated genes confirmed a proinflammatory state highlighting activation of endolysosomal pathways, secretion and immune cell degranulation previously associated with myeloid cells in ALS 45 (Fig. 6g,h ). Further subclustering identified three groups: homeostatic Micro0, ‘disease-associated microglia (DAM)’-like Micro1 and cycling Micro2 (Extended Data Fig. 9f–h ). Notably, genes that characterized Micro1 were also upregulated in sALS (Extended Data Fig. 9i,j ), in conjunction with a downregulation of homeostatic genes and upregulation of reactive pathways (Extended Data Fig. 9k–m ).

a , t SNE projection of microglia (ALS n  = 759 nuclei and control n  = 693 nuclei). b , c , Volcano plot of genes upregulated in microglia from ALS. Genes identified in GO terms for endocytosis and exocytosis ( b ) and genes associated with neurodegenerative diseases ( c ). d , Violin plots of representative genes upregulated in patients with ALS associated with reactive microglia (geometric box plots represent median and interquantile ranges; symbols indicate log 2 (average expression) per individual) (fraction of cell expressing). e , A dot plot representing expression of genes associated with ALS–FTD pathogenesis. f , Violin plots of representative ALS–FTD genes upregulated in ALS (geometric box plots represent median and interquantile ranges; symbols indicate log 2 (average expression) per individual) (fraction of cell expressing). g , GO analysis for genes upregulated in ALS microglia, highlighted terms involved in myeloid cells biology and/or pathogenesis of ALS (WP, WikiPathways). h , Violin plots representing z -scores for selected, statistically significant GO terms from f (geometric box plots represent median and interquantile ranges; symbols indicate average score per individual). i , Comparison of genes upregulated in microglia from ALS with genes upregulated in microglia in other neurodegenerative diseases (for a – i , n  = 3 control individuals and n  = 5 patients with sALS). nUMI, normalized Unique Molecular Identifier.

To identify modulators of this signature, we used the Connectivity Map pipeline 46 , which contains gene expression data of nine human cell lines after thousands of perturbations, allowing association between a given transcriptomic signature and a specific alteration. This analysis revealed that genes dysregulated in microglia positively correlated with regulators of cell cycle and senescence, suggesting an exhaustion of microglial proliferation. We also found a negative correlation with a type I interferon-associated responses ( IFNB1 ), often targeted in treatments for neurological diseases to reduce inflammation (Extended Data Fig. 10a ).

By comparing our results with published snRNAseq studies 35 , 47 , we identified dysregulation of lipid metabolism ( APOE , APOC1 and SPP1 ) as a common feature in microglia, genes associated with DAMs shared between ALS and MS ( CTSD , GPNMB , CPM and LPL ) and ALS and AD (for example, TREM2 ), as inferred by bulk RNA sequencing studies 7 (Fig. 5i ). Genes specifically upregulated in ALS were related to vesicle trafficking, myeloid cell degranulation and the lysosome (for example, SQSTM1/p62 , LGALS3 , GRN , ASAH1 and LRRK2 ). This evidence suggests the induction of a shared microglial reactive state, yet in ALS these changes are connected to endolysosomal pathways.

Given the stress signature identified in neurons, we wondered whether these transcriptomic changes were driven by neuronal distress. We differentiated induced microglia-like cells (iMGLs) 48 and neurons 38 , 49 from hPS cells, triggered neuronal apoptosis and then introduced apoptotic neurons to iMGLs in vitro 48 (Fig. 7a,b and Extended Data Fig. 10b ). Quantitative assessment by quantitative reverse trascription polymerase chain reaction (RT–qPCR) confirmed the treatment lead to significant downregulation of homeostatic genes, upregulation of genes involved in endolysosomal trafficking ( CTSD , ITGAX , LGALS3 and SQSTM1/p62 ) and downregulation of actively cycling cells markers (Fig. 7c,d and Extended Data Fig. 10c ), suggesting that changes identified in microglia from patients are, at least in part, a response to neuronal apoptosis.

a , Diagram of microglia and neuronal differentiation from PSCs and induction of apoptosis neurons and feeding to iMGLs (piNs, patterned induced neurons). b , Brightfield images of untreated day 40 iMGLs and day 40 iMGLs that were fed apoptotic neurons for 24 h. c . RT–qPCR quantification of homeostatic and DAM genes after feeding (AN, apoptotic neurons). d , RT–qPCR quantification of selected ALS–FTD-associated and lysosomal genes 24 h after feeding iMGLs with apoptotic neurons (AN, apoptotic neurons). ( t -test, * P  < 0.05, ** P  < 0.01, *** P  < 0.001, n  = 3 biological replicates). piNS, induced neurons from hPS cells.

A key question in the study of neurodegeneration is why certain cell types are more susceptible to different diseases. In this study, we identified the enrichment for ALS–FTD-associated genes in a class of ETNs, which provides a connection between this neuronal subtype and its propensity to accumulate TDP-43 aggregates 31 leading to their gradual loss in ALS–FTD 10 . This enrichment is not recapitulated for risk factors connected to AD and MS, related to immune processes and more enriched in microglia 18 , 19 , 20 . One study suggested that ALS-associated variants connected to autophagy and protein clearing are most highly expressed in glutamatergic neurons 33 , and these findings add to the importance of axonal dynamics and ribonucleotide metabolism 34 ; here, we provide a more detailed dissection of which subtype that might be.

Additionally, we identified a broadly shared transcriptomic signature of cellular stress pathways in classes of deep-layer excitatory neurons. These alterations in RNA translation and proteostasis have been implicated in models of ALS 1 , 2 ; our study highlights their cell type specificity and links them to rare mutations in regulators of these pathways in familial forms of ALS 4 . These molecular mechanisms are confirmed to be connected to proteasomal function and proteostasis by human neuronal in vitro models, a system that is already being used to identify therapeutic candidates funneled into clinical trial pipelines 50 . The nuclear nature and the low coverage of this kind of sequencing methodology, but also the small sample size in our study, obviates further, confident dissection of the neuronal-specific changes in RNA biology identified in in vitro models and patient samples 37 . Moreover, the phenotypic and potentially genetic heterogeneity of sporadic ALS is probably reflected in our dataset and should be considered. This study is not powered to distinguish the relative contribution of technical effects versus intrinsic heterogeneity across individuals with sALS, but it opens the field to important considerations for future studies that should include greater numbers of patients. Nonetheless, our report highlights the importance of RNA metabolism and proteostasis and their specific misregulation in ETNs. Mouse models where these pathways are specifically altered in CSMNs might shed a light on their interplay in this specific neuronal type.

We suggest two mechanisms by which ETNs are rendered more susceptible to sALS: (1) the intrinsically higher expression of risk factors and (2) processes of degeneration in classes of ETNs that might exacerbate/contribute to their vulnerability in a combinatorial effect. Recent snRNAseq studies unraveled susceptibility of specific neuronal types in other diseases: mid-layer RORB neurons in AD 51 , 52 , upper-layer CUX2 neurons in MS 22 , dopaminergic neurons in Parkinson’s disease 53 and ETNs affected in ALS–FTD as described by our study and spinal cord motor neurons as recently suggested in ALS 54 . Impairment of proteostatic mechanisms seems to be a common theme in degenerating neurons regardless of the disease; however, only in ALS are these changes specifically connected to upregulation of transcripts connected to RNA metabolism, a trend that appears to go in the opposite direction in AD 52 . Integrative analyses of these studies might mark the beginning of a new understanding of the mechanisms behind selective neuronal vulnerability to different diseases.

Emerging studies have shown that glial cells are important modifiers in ALS–FTD 44 . We show that changes in processes involved in oligodendrocyte differentiation and myelination may contribute to degeneration and/or be a coordinated response to ALS and appear to contrast with those described in MS 43 . We revealed perturbations in key myelin regulators, such as OPALIN , CNP and MAG , across oligodendrocyte clusters but in these cells only, as opposed to AD where myelination-related changes were present across multiple cell types 35 , 55 . Given the similarities in the stress signature identified in neurons with changes in MS lesions but not in AD, it is puzzling how changes in myelination might be a consequence or cause of neuronal degeneration.

Intriguingly, recent work showed expression of neuronal RNA in oligodendrocytes in human motor cortex 24 . Upregulation of synaptic transcripts in this cell type in patients with sALS might represent phagocytic activity 56 or the need for synaptic proteins during deposition of myelin sheath 57 . These speculations are interesting if coupled with the upregulation of synaptic machinery in upper layer neurons and the documented loss of postsynaptic molecules in ETNs in ALS 58 . Recent snRNAseq studies of FTD cortices identified changes in myelinating cells in response to neuronal loss and underlined the importance of cell-to-cell communication 40 , also GWAS studies have pointed at excitatory neurons, myelinating cells and inhibitory neurons’ sensitivity to genetic risks for ALS 34 . These observations suggest a coordinated response of the motor circuit in an attempt to compensate for loss of inputs to the cord. Further investigations could focus on shifting oligodendroglial states in mouse models and determine changes during disease progression to complement efforts aimed at controlling neuronal activity 59 .

Finally, we found distinct perturbations in ALS-associated microglia, particularly in endolysosomal pathways. We and others have implicated ALS–FTD-associated gene C9orf72 in endosomal trafficking and secretion in myeloid cells 44 , 45 and the upregulation of lysosomal constituents, for example, CTSD , was identified in this study and by others in patients 60 . Coupled with the upregulation of ALS–FTD-associated genes SQSTM1/p62 , OPTN , TREM2 and GRN , this suggests a mechanistic convergence on vesicle trafficking and inflammatory pathways that may initiate/exacerbate the homeostatic-to-DAM transition in ALS 7 . The interferon response-related changes we delineate, as identified by others in C9orf72 ALS 61 , provide a parallel between sporadic and familial ALS. Overall, these changes had partial overlap with microglia in AD 35 and in MS 47 , suggesting that drugs modulating myeloid cells in neurodegenerative diseases may provide a basis for new therapeutic approaches. Recent reports showed how DAM might be beneficial in disease contexts 62 , but others have inferred that microglial activation might result in poor disease outcomes. Studies manipulating microglial states might elucidate their ‘friend or foe’ role in sALS 48 .

In summary, we show that classes of ETNs require the expression of a collection of genetic risk factors for ALS–FTD with pivotal roles in proteostasis. This intrinsically higher expression of disease-associated genes might be at the bottom of a ‘first over the line’ mechanism leading to disruptions in groups of deep layer excitatory neurons. These alterations trigger a cascade of responses in glia: oligodendroglia shift from a myelinating to a neuronally engaged state and microglia activate a proinflammatory signature. Our study offers a view in which neurocentric disease vulnerability might spark responses in other cell types, but it also shows that enrichment of ALS–FTD-related genes in ETNs is coupled with processes engaging these genes in other cells too, that is, microglia. This view is a first insight into the disruptions of cortical biology in ALS and provides a connection between changes in cellular components and mechanisms associated with ALS. Future investigations should consider multicellular disruptions in ALS–FTD, where the survival of the neuron is unmistakably pivotal, but targeting other cells to reduce inflammation, promote myelination and bolster neuronal circuitry may re-establish a neuroprotective environment.

Limitations of this study

One limitation is the small size of the cohort. ALS is a heterogeneous disease 7 and smaller cohort sizes might not fully recapitulate its etiological diversity. Only recently have biobanks started to collect enough samples 6 , 7 , 8 , and we hope that increased sample availability and affordability of single-nucleus technology will allow a more comprehensive view of alterations in ALS. A larger cohort would also enable a more stringent analysis of differentially expressed transcripts that incorporates more sophisticated analytical tool.

We recognize that consensus on best practices in snRNAseq is still being reached, including for re-analysis of published studies 63 , 64 . We acknowledge that using single cells as a variable and not pseudobulked individuals might yield larger gene sets with possible confounding factors. We also recognize that our additional analyses revealed that some specific genes might be derived from outlier-driven effects and because of that we limited follow-up analyses only to genes shared by at least four out of five individuals. This is why we highlighted common features in wider biological pathways disrupted in cell types in patients with ALS followed up by validations at the protein level in a separate cohort of patients’ samples or in our in vitro studies. We hope that new reports will take into account the need for a more stringent investigation of reproducibility and we advocate for a more transparent conveying of results.

We also recognize that our study would benefit from additional validation at the RNA and/or protein level. This would elucidate some of the intriguing questions we raised. For example, are oligodendrocytes really expressing higher levels of neuronal genes or is this an artifact 65 ? Nonetheless, we believe that this study provides novel insights in the involvement of different cell types in ALS and a different view in the motor cortex of patients with ALS. The increase in cohort sizes, more sophisticated analyses and new technologies, such as spatial transcriptomics, might further enrich the understanding of neurodegeneration in ALS.

Human donor tissue sources and ethics

Frozen postmortem human cortical samples from cases of patients with sporadic ALS and age-matched controls were obtained from the Target ALS Neuropathology Core that drew upon the repositories of five institutions. Specimens from the medial, lateral or unspecified motor cortex were grouped together. Deidentified postmortem brain tissue samples were obtained from the Massachusetts Alzheimer’s Disease Research Center at Massachusetts General Hospital (MGH) and the Target ALS Multicenter Human Postmortem Tissue Core, which integrates five academic tissue repositories for ALS research. The protocols of the Massachusetts Alzheimer’s Disease Research Center for brain donations and the collection of postmortem tissue and clinical information for research purposes were approved by the Institutional Review Board of Partners Healthcare (currently Mass General Brigham) at MGH. Informed consent for brain autopsy and the use of postmortem tissue for research was provided by the legal next-of-kin in compliance with local and institutional guidelines at all brain tissue repositories involved. Use of postmortem deidentified tissue samples followed ‘Not Human Subjects Research’ determinations by the Harvard Faculty of Arts and Sciences and Partners (MGH) and considered exempt from the instituational research board given lack of interaction with living individuals/participants. The study protocol was further approved by Harvard Stem Cell and Regenerative Biology Department, Harvard University. Informed consent and study protocol for human stem cell work were provided by Stanley Center for Psychiatric Research at Broad Institute of MIT and Harvard and the Harvard Stem Cell and Regenerative Biology Department at Harvard University.

Isolation of nuclei

RNA quality of brain samples was assessed by running bulk nuclear RNA on an Agilent TapeStation for RNA integrity number scores. Extraction of nuclei from frozen samples was performed as previously described 66 . Briefly, tissue was dissected and minced with a razor blade on ice and then placed in 4 ml ice-cold extraction buffer (wash buffer (82 mM Na2SO4, 30 mM K2SO4, 5 mM MgCl2, 10 mM glucose and 10 mM HEPES, pH adjusted to 7.4 with NaOH) containing 1% Triton X-100 and 5% Kollidon VA64). Tissue was homogenized with repeated pipetting, followed by passing the homogenized suspension twice through a 26.5 gauge needle on a 3 ml syringe (prechilled), once through a 20 mm mesh filter and once through a 5 mm filter using vacuum. The nuclei were then diluted in 50 ml ice-cold wash buffer, split across four 50 ml tubes and centrifuged at 500 g for 10 min at 4 °C. The supernatant was discarded, the nuclei pellet was resuspended in 1 ml cold wash buffer.

10× loading and library preparation

Nuclei were 4,6-diamidino-2-phenylindole (DAPI) stained with Hoechst, loaded onto a hemocytometer and counted using brightfield and fluorescence microscopy. The solution was diluted to ~176 nuclei μl −1 before proceeding with Drop-seq, as described 14 . Complementary DNA amplification was performed using around 6,000 beads per reaction with 16 PCR cycles. The integrity of both the complementary DNA and tagmented libraries were assessed for quality control on the Agilent Bioanalyzer. Libraries were sequenced on a Nova-seq S2, with a 60 bp genomic read. Reads were aligned to the human genome assembly (hg19). Digital gene expression files were generated with the Zamboni drop-seq analysis pipeline, designed by the McCarroll group 66 , 67 .

Filtering of expression matrices and clustering of single nuclei

A single matrix for all samples was built by filtering any barcode with less than 400 genes and resulting in a matrix of 27,600 genes across 119,510 barcodes. This combined UMI matrix was used for downstream analysis using Seurat (v3.0.2) 15 . A Seurat object was created from this matrix by setting up a first filter of min.cells=20 per genes. After that, barcodes were further filtered by number of genes detected nFeature_RNA > 600 and nFeature_RNA < 6,000. Distribution of genes and UMIs were used as parameters for filtering barcodes. The matrix was then processed via the Seurat pipeline: log normalized by a factor of 10,000, followed by regressing UMI counts (nCount_RNA) and scaled for gene expression.

After quality filtering, 79,830 barcodes and 27,600 genes were used to compute shared nearest-neighbor graphs and t -distributed stochastic neighbor embedding ( t SNE) projections using the first ten statistically significant principal components. As previously described 44 , 68 , t SNE projection was used to determine minimum number of clusters at Resolution=0.2 (FindClusters). Broad cellular identities were assigned to groups on the basis of DEGs as calculated by Wilcoxon rank sum test in FindAllMarkers(min.pct=0.25, logfc.threshold=0.25). One subcluster with a specifically high ratio of UMIs/genes was filtered out resulting in 79,169 barcodes grouped in seven major cell types. Markers for specific cell types were identified in previously published small conditional RNA sequencing studies 16 .

Analysis of cellular subtypes was conducted by subsetting each group. Isolated barcodes were renormalized and scaled and relevant principal components were used for clustering as a separate analysis. Newly scaled matrices were used for DGE analysis with MAST algorithm in Seurat, as previously reported 38 , 43 , 44 , 53 , 68 , with parameters FindAllMarkers(min.pct=0.10, logfc.threshold=0.25) and subclustering for identification of subgroups. DGEs for downsampled excitatory neurons groups were computed by adding parameters to the functions described above, FindAllMarkers(min.pct=0.10, logfc.threshold=0.25, min.cells = ‘minimum number in the group’, random.seed=TRUE), and re-iterative lists were generated with >95% of overlap (data not shown). PCA by individual was performed using matrices generated by AverageExpression(celltype, group.by = ”ID”, return.seurat=TRUE); these were normalized and scaled and PCA was run using the DGEs with RunPCA(celltype.averages, features=c(DGEs), npcs=7). To further confirm that changes in DGEs were not driven by one individual only, we ran DGE analysis by excluding cells from one ALS individual at a time (control versus ALS-1) in a re-iterative manner and showed that magnitude of changes in DGE expression were still similar and that identified DGEs were shared by all five or at least four out of five individuals; these gene lists were used for GOy and protein–protein interaction analyses. Gene scores for different cellular subclusters were computed in each renormalized, rescaled submatrix using the AddModuleScore function in Seurat v3.0.2.

Re-analysis of publicly available datasets was performed using matrices and metadata available. Only barcodes with available metadata concerning their cellular identity were selected to use identities assigned by peer review publication 22 , 30 . The available barcodes were then loaded into Seurat v4.0.1 (ref. 69 ). Gene scores for different cellular subclusters were computed in each renormalized, rescaled submatrix using the AddModuleScore function, as previously described. Re-analysis of spatial transcriptomic from Maynard et al. was performed using publicly available data and codes from publication itself 29 .

GO, interactome and GSEAs

For GO terms analysis, we selected statistically significant upregulated or downregulated genes identified in each subcluster as described before (adjusted P values <0.05, log fold change (FC) of 2). These lists were fed in the gProfiler pipeline 70 with the following settings: use only annotated genes, g:SCS threshold of 0.05, GO cellular components and GO biological processes (26 May 2020 to 9 December 2021), only statistically significant pathways are highlighted. Only statistically significant upregulated genes identified in each subcluster as described before (adjusted P values < 0.05, log FC of 2) were used for GO analysis. The interactome map was built using STRING 71 protein–protein interaction networks, all statistically significant upregulated genes were used, 810 were identified as interacting partners using ‘experiments’ as interaction sources and a medium confidence threshold (0.400), only interacting partners are shown in Extended Data Fig. 6 . Gene set enrichment analysis (GSEA) was performed using GSEA software designed by UC San Diego and the Broad Institute (v4.0.3) 72 . Briefly, gene expression matrices were generated in which for each subcluster each individual was a metacell, lists for disease-associated risk genes were compiled using available datasets (PubMed, ALS–FTD; Source Data Table 2 ) or recently published GWAS for AD 18 , 19 and MS 20 .

Generation of microglia-like cells

Microglial-like cells were differentiated as described 48 . Briefly, WA01-H1 hPS cells were cultured in E8 medium (Stemcell Technologies) on Matrigel (Corning), dissociated with accutase (Stemcell Technologies), centrifuged at 300 g for 5 min and resuspended in E8 medium with 10 μM Y-27632 ROCK inhibitor, with 2 M cells transferred to a low-attachment T25 flask in 4 ml of medium and left in the suspension for 24 h. The first 10 days of differentiation are carried out in iHPC medium: Iscove’s modified Dulbecco’s medium (50%, Stemcell Technologies), F12 (50%, Stemcell Technologies), ITS-G-X 2% vol/vol (Thermo Fisher), l -ascorbic acid 2-phosphate (64 μg ml −1 , Sigma), monothioglycerol (400 mM, Sigma), PVA (10 mg ml −1 ; Sigma), glutamax (1×, Stemcell Technologies), chemically defined lipid concentrate (1×, Stemcell Technologies) and nonessential amino acids (Stemcell Technologies). After 24 h (day 0), cells were collected, and differentiation is started in iHPC medium supplemented with fibroblast growth factor 2 (FGF2) (PeproTech, 50 ng ml −1 ), bone morphogenetic protein (PeproTech, 50 ng ml −1 ), activin A (PeproTech, 12.5 ng ml −1 ), Y-27632 ROCK inhibitor (1 μM) and LiCl (2 mM) and transferred in a hypoxic incubator (20% O 2 , 5% CO 2 at 37 °C). On day 2, the medium was changed to iHPC medium plus FGF2 (PeproTech, 50 ng ml −1 ) and vascular enothelial growth factor (PeproTech, 50 ng ml −1 ) and returned to hypoxic conditions. On day 4, cells were resuspended in iHPC medium supplemented with FGF2 (PeproTech, 50 ng ml −1 ), vascular endothelial growth factor (PeproTech, 50 ng ml −1 ), TPO (PeproTech, 50 ng ml −1 ), SCF (PeproTech, 10 ng ml −1 ), interleukin (IL)-6 (PeproTech, 50 ng ml −1 ) and IL-3 (PeproTech, 10 ng ml −1 ), and placed into a normoxic incubator (20% O 2 , 5% CO 2 at 37 °C). Expansion of hematopoietic progenitors was continued by supplementing the flasks with 1 ml of iHPC medium with small molecules every 2 days. On day 10, cells were collected and filtered through a 40 mm filter. The single-cell suspension was counted and plated at 500,000 cells per well in a six-well plate coated with Matrigel (Corning) in microglia differentiation medium: Dulbecco’s modified Eagle medium/F12 (Stemcell Technologies), ITS-G 2% vol/vol (Thermo Fisher Scientific), B27 (2% vol/vol, Stemcell Technologies), N 2 (0.5% vol/vol, Stemcell Technologies), monothioglycerol (200 mM, Sigma), glutamax (1×, Stemcell Technologies), nonessential amino acids (1×, Stemcell Technologies), supplemented with macrophage colony-stimulating factor (25 ng ml −1 , PeproTech), IL-34 (100 ng ml, PeproTech) and ransforming growth factor β-1 (50 ng ml −1 , PeproTech). iMGLs were kept in this medium for 20 days with change three times a week. On day 30, cells were collected and plated on poly- d -lysine/laminin-coated dishes in microglia differentiation medium supplemented with CD200 (100 ng ml −1 , Novoprotein) and CX3CL1 (100 ng ml −1 , PeproTech), macrophage colony-stimulating factor (25 ng ml, PeproTech), IL-34 (100 ng/ml, PeproTech) and transforming growth factor β-1 (50 ng ml −1 , PeproTech) until day 40.

Feeding of apoptotic neurons to microglia-like cells

For feeding assays, neurons were generated from human iPSCs using an NGN2 overexpression system, as described previously 38 , 49 , 73 . Day-30 WA01-H1 hiPSC-derived neurons were treated with 2 μM H 2 O 2 for 24 h to induce apoptosis. Apoptotic neurons were gently collected from the plate and the medium containing the apoptotic bodies was transferred into wells containing day 40 iMGLs. After 24 h, iMGLs subjected to apoptotic neurons and controls were collected for RNA extraction.

RNA extraction and RT–qPCR analysis

RNA was extracted with the miRNeasy Mini kit (Qiagen, 217004). cDNA was produced with iScript kit (Bio-Rad) using 50 ng of RNA. RT–qPCR reactions were performed in triplicates using 20 ng of cDNA with SYBR Green (Bio-Rad) and were run on a CFX96 Touch PCR Machine for 39 cycles at 95 °C for 15 s, 60 °C for 30 s and 55 °C for 30 s.

Generation of hiPSC-derived neurons for bulk RNA sequencing

Human embryonic stem cells were cultured in mTeSR (Stemcell Technologies) on Matrigel (Corning). Motor neurons were generated from HuES-3-Hb9:GFP based on the differentiation protocol previously described 74 . On completion of differentiation, cells were sorted via flow cytometry based on green fluorescent protein (GFP) signal intensity to yield GFP-positive neurons that were plated on PDL/laminin-coated plates (Sigma, Life Technologies). Neurons were maintained in neurobasal medium (Life Technologies) and supplemented with N 2 (STEMCELL Technologies), B27 (Life Technologies), glutamax (Life Technologies), nonessential amino acids (Life Technologies) and neurotrophic factors (BDNF, GDNF and CNTF), and were grown for 28 days before the application of the proteasome inhibitors MG132 for 24 h.

RNA was extracted using RNeasy Plus kit (Qiagen), libraries were prepared using the Illumina TruSeq RNA kit v2 according to the manufacturer’s directions and sequenced at the Broad Institute core with samples randomly assigned between two flow chambers. The total population RNA sequencing FASTQ data was aligned against ENSEMBL human reference genome (build GRCh37/hg19) using STAR (v.2.4.0). Cufflinks (v.2.2.1) was used to derive normalized gene expression in fragments per kilo base per million. The read counts were obtained from the aligned BAM-files in R using Rsubread 73 . Differential gene expression was analyzed from the read counts in DESeq2 using a Wald’s test for the treatment dosage and controlling for the sequencing flow cell 73 .

Western blot analysis

As previously described, tissue was minced, lysed in RIPA buffer with protease inhibitors (Roche) and sonicated 75 . After centrifugation, the supernatant was collected as soluble fraction and the insoluble pellet was resuspended in 8 M urea buffer (Bio-Rad, 1632103). After protein quantification by the bicinchoninic acid assay (Thermo Fisher), 10 µg of protein was preheated in Laemmli’s buffer (Bio-Rad) and loaded in 4–20% mini-PROTEAN TGX precast protein gels (Bio-Rad), and then gels were transferred to a polyvinylidene fluoride membrane. Membranes were blocked in Odyssey blocking buffer (Li-Cor) and incubated overnight at 4 °C with primary antibodies. After washing with TBS-T, membranes were incubated with IRDye secondary antibodies (1:10,000, Li-Cor) for 1 h and imaged with the Odyssey CLx imaging system (Li-Cor). Primary antibodies used 1:1,000 dilution: TDP-43 (PeproTech 10782-2-AP), GAPDH (Millipore cat. no. MAB374; CST 2118 (14C10)), MBP (Thermo Fisher PA-1-10008), CNP (Abcam ab6319(11-5b)), 20S (Enzo BML-PW8195-0025) and ubiquitin (CST 3936 T (P4D1)). IRDye provided by Licor was used at 1:10,000 dilution.

Immunofluorescence assays

Cells were washed once with phosphate-buffered saline (PBS), fixed with 4% paraformaldehyde for 20 min, washed again in PBS and blocked for 1 h in 0.1% Triton in PBS with 10% donkey serum. Fixed cells were then washed and incubated overnight with primary antibodies at 4 °C. Primary antibody solution was washed and cells were subsequently incubated with secondary antibodies (1:2,000, AlexaFluor, Life Technologies) at room temperature for 1 h, washed with PBS and stained with DAPI. Primary antibodies used were Tuj1 (1:250, R&D, MAB1195) and TDP-43 (1:200, PeproTech 10782-2-AP). Images were analyzed using FIJI.

Proteasome activity assay

Neurons were sorted in 96-well plates and, after 2 weeks of maturation, treated for 24 h. Cells were washed with 1× PBS, exposed to ProteasomeGlo (Promega, G8660) and incubated for 30 min at room temperature. Fluorescence was measured using a Cytation 3 reader (BioTek).

Statistics and reproducibility

No statistical method was used to predetermine sample size. No statistical methods were used to predetermine sample sizes but our sample sizes are similar to those reported in previous publications 22 , 24 , 26 , 29 , 30 , 40 , 43 , 44 , 47 , 48 , 51 , 52 , 54 , 68 . No data were excluded from the analyses. The experiments were not randomized. The investigators were not blinded to allocation during experiments and outcome assessment. Data distribution was assumed to be normal but this was not formally tested. Data collection and analysis were not performed blind to the conditions of the experiments. Software used for analyses were Licor (Image Studio version 2.1), GraphPad Prism (version 7 and above), ImageJ (FIJI version 2.14) with Nikon NIS Elements version 4.0, Seurat version 3.0.2 and version 4.0.1, and GSEA version 4.0.3.

Reporting summary

Further information on research design is available in the Nature Portfolio Reporting Summary linked to this article.

Data availability

Transcriptomic raw data have been deposited in the Gene Expression Omnibus database under accession number GSE226753. All other data supporting the findings of this study are available as source data files or from the corresponding author upon reasonable request. All other data analyzed from previously published sources will be available at publication references in the manuscript (for Schirmer et al. Sequence Read Archive (SRA), under accession number PRJNA544731 and NCBI Bioproject ID: 544731; for Velmeshev et al. Sequence Read Archive, accession number PRJNA434002 ; for Maynard et al. available via GitHub at https://github.com/LieberInstitute/HumanPilot and https://github.com/LieberInstitute/spatialLIBD ).

Taylor, J. P., Brown, R. H. Jr. & Cleveland, D. W. Decoding ALS: from genes to mechanism. Nature 539 , 197–206 (2016).

Article   PubMed   PubMed Central   Google Scholar  

Brown, R. H. & Al-Chalabi, A. Amyotrophic lateral sclerosis. N. Engl. J. Med. 377 , 162–172 (2017).

Article   CAS   PubMed   Google Scholar  

Giacomelli, E. et al. Human stem cell models of neurodegeneration: from basic science of amyotrophic lateral sclerosis to clinical translation. Cell Stem Cell 29 , 11–35 (2022).

Article   CAS   PubMed   PubMed Central   Google Scholar  

Mordes, D. A. et al. Dipeptide repeat proteins activate a heat shock response found in C9ORF72-ALS/FTLD patients. Acta Neuropathol. Commun. 6 , 55 (2018).

D’Erchia, A. M. et al. Massive transcriptome sequencing of human spinal cord tissues provides new insights into motor neuron degeneration in ALS. Sci. Rep. 7 , 10046 (2017).

Tam, O. H. et al. Postmortem cortex samples identify distinct molecular subtypes of ALS: retrotransposon activation, oxidative stress, and activated glia. Cell Rep. 29 , 1164–1177 e1165 (2019).

Humphrey, J. et al. Integrative transcriptomic analysis of the amyotrophic lateral sclerosis spinal cord implicates glial activation and suggests new risk genes. Nat. Neurosci. 26 , 150–162 (2023).

Eshima, J. et al. Molecular subtypes of ALS are associated with differences in patient prognosis. Nat. Commun. 14 , 95 (2023).

Neumann, M. et al. Ubiquitinated TDP-43 in frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Science 314 , 130–133 (2006).

Hammer, R. P. Jr., Tomiyasu, U. & Scheibel, A. B. Degeneration of the human Betz cell due to amyotrophic lateral sclerosis. Exp. Neurol. 63 , 336–346 (1979).

Article   PubMed   Google Scholar  

Seeley, W. W. et al. Early frontotemporal dementia targets neurons unique to apes and humans. Ann. Neurol. 60 , 660–667 (2006).

Kang, S. H. et al. Degeneration and impaired regeneration of gray matter oligodendrocytes in amyotrophic lateral sclerosis. Nat. Neurosci. 16 , 571–579 (2013).

Boillee, S. et al. Onset and progression in inherited ALS determined by motor neurons and microglia. Science 312 , 1389–1392 (2006).

Macosko, E. Z. et al. Highly parallel genome-wide expression profiling of individual cells using nanoliter droplets. Cell 161 , 1202–1214 (2015).

Stuart, T. et al. Comprehensive integration of single-cell data. Cell 177 , 1888–1902 e1821 (2019).

Lake, B. B. et al. Integrative single-cell analysis of transcriptional and epigenetic states in the human adult brain. Nat. Biotechnol. 36 , 70–80 (2018).

Tirosh, I. et al. Dissecting the multicellular ecosystem of metastatic melanoma by single-cell RNA-seq. Science 352 , 189–196 (2016).

Kunkle, B. W. et al. Genetic meta-analysis of diagnosed Alzheimer’s disease identifies new risk loci and implicates Abeta, tau, immunity and lipid processing. Nat. Genet. 51 , 414–430 (2019).

Jansen, I. E. et al. Genome-wide meta-analysis identifies new loci and functional pathways influencing Alzheimer’s disease risk. Nat. Genet. 51 , 404–413 (2019).

International Multiple Sclerosis Genetics Consortium Multiple sclerosis genomic map implicates peripheral immune cells and microglia in susceptibility. Science 365 , eaav7188 (2019).

Article   PubMed Central   Google Scholar  

Arlotta, P. et al. Neuronal subtype-specific genes that control corticospinal motor neuron development in vivo. Neuron 45 , 207–221 (2005).

Schirmer, L. et al. Neuronal vulnerability and multilineage diversity in multiple sclerosis. Nature 573 , 75–82 (2019).

Ozdinler, P. H. et al. Corticospinal motor neurons and related subcerebral projection neurons undergo early and specific neurodegeneration in hSOD1G(9)(3)A transgenic ALS mice. J. Neurosci. 31 , 4166–4177 (2011).

Bakken, T. E. et al. Comparative cellular analysis of motor cortex in human, marmoset and mouse. Nature 598 , 111–119 (2021).

Guerra San Juan, I. et al. Loss of mouse Stmn2 function causes motor neuropathy. Neuron https://doi.org/10.1016/j.neuron.2022.02.011 (2022).

Hodge, R. D. et al. Transcriptomic evidence that von Economo neurons are regionally specialized extratelencephalic-projecting excitatory neurons. Nat. Commun. 11 , 1172 (2020).

Cobos, I. & Seeley, W. W. Human von Economo neurons express transcription factors associated with layer V subcerebral projection neurons. Cereb Cortex 25 , 213–220 (2015).

Zeng, H. et al. Large-scale cellular-resolution gene profiling in human neocortex reveals species-specific molecular signatures. Cell 149 , 483–496 (2012).

Maynard, K. R. et al. Transcriptome-scale spatial gene expression in the human dorsolateral prefrontal cortex. Nat. Neurosci. 24 , 425–436 (2021).

Velmeshev, D. et al. Single-cell genomics identifies cell type-specific molecular changes in autism. Science 364 , 685–689 (2019).

Nana, A. L. et al. Neurons selectively targeted in frontotemporal dementia reveal early stage TDP-43 pathobiology. Acta Neuropathol. 137 , 27–46 (2019).

Porta, S. et al. Patient-derived frontotemporal lobar degeneration brain extracts induce formation and spreading of TDP-43 pathology in vivo. Nat. Commun. 9 , 4220 (2018).

van Rheenen, W. et al. Common and rare variant association analyses in amyotrophic lateral sclerosis identify 15 risk loci with distinct genetic architectures and neuron-specific biology. Nat. Genet. 53 , 1636–1648 (2021).

Saez-Atienzar, S. et al. Genetic analysis of amyotrophic lateral sclerosis identifies contributing pathways and cell types. Sci. Adv. 7 , eabd9036 (2021).

Mathys, H. et al. Single-cell transcriptomic analysis of Alzheimer’s disease. Nature 570 , 332–337 (2019).

Limone, F., Klim, J. R. & Mordes, D. A. Pluripotent stem cell strategies for rebuilding the human brain. Front. Aging Neurosci. 14 , 1017299 (2022).

Klim, J. R. et al. ALS-implicated protein TDP-43 sustains levels of STMN2, a mediator of motor neuron growth and repair. Nat. Neurosci. 22 , 167–179 (2019).

Limone, F. et al. Efficient generation of lower induced motor neurons by coupling Ngn2 expression with developmental cues. Cell Rep. 42 , 111896 (2023).

Tomassy, G. S. et al. Distinct profiles of myelin distribution along single axons of pyramidal neurons in the neocortex. Science 344 , 319–324 (2014).

Gerrits, E. et al. Neurovascular dysfunction in GRN-associated frontotemporal dementia identified by single-nucleus RNA sequencing of human cerebral cortex. Nat. Neurosci. 25 , 1034–1048 (2022).

Giera, S. et al. The adhesion G protein-coupled receptor GPR56 is a cell-autonomous regulator of oligodendrocyte development. Nat. Commun. 6 , 6121 (2015).

Yang, H. J., Vainshtein, A., Maik-Rachline, G. & Peles, E. G protein-coupled receptor 37 is a negative regulator of oligodendrocyte differentiation and myelination. Nat. Commun. 7 , 10884 (2016).

Jakel, S. et al. Altered human oligodendrocyte heterogeneity in multiple sclerosis. Nature 566 , 543–547 (2019).

Limone, F. et al. Myeloid and lymphoid expression of C9orf72 regulates IL-17A signaling in mice. Sci. Transl. Med. 16 , eadg7895 (2024).

Burberry, A. et al. C9orf72 suppresses systemic and neural inflammation induced by gut bacteria. Nature 582 , 89–94 (2020).

Subramanian, A. et al. A next generation connectivity map: L1000 platform and the first 1,000,000 profiles. Cell 171 , 1437–1452 e1417 (2017).

Masuda, T. et al. Spatial and temporal heterogeneity of mouse and human microglia at single-cell resolution. Nature 566 , 388–392 (2019).

Dolan, M. J. et al. Exposure of iPSC-derived human microglia to brain substrates enables the generation and manipulation of diverse transcriptional states in vitro. Nat. Immunol. 24 , 1382–1390 (2023).

Nehme, R. et al. Combining NGN2 programming with developmental patterning generates human excitatory neurons with NMDAR-mediated synaptic transmission. Cell Rep. 23 , 2509–2523 (2018).

Morimoto, S. et al. Phase 1/2a clinical trial in ALS with ropinirole, a drug candidate identified by iPSC drug discovery. Cell Stem Cell 30 , 766–780 e769 (2023).

Leng, K. et al. Molecular characterization of selectively vulnerable neurons in Alzheimer’s disease. Nat. Neurosci. https://doi.org/10.1038/s41593-020-00764-7 (2021).

Otero-Garcia, M. et al. Molecular signatures underlying neurofibrillary tangle susceptibility in Alzheimer’s disease. Neuron 110 , 2929–2948 e2928 (2022).

Kamath, T. et al. Single-cell genomic profiling of human dopamine neurons identifies a population that selectively degenerates in Parkinson’s disease. Nat. Neurosci. 25 , 588–595 (2022).

Yadav, A. et al. A cellular taxonomy of the adult human spinal cord. Neuron 111 , 328–344 e327 (2023).

Sadick, J. S. et al. Astrocytes and oligodendrocytes undergo subtype-specific transcriptional changes in Alzheimer’s disease. Neuron 110 , 1788–1805 e1710 (2022).

Falcao, A. M. et al. Disease-specific oligodendrocyte lineage cells arise in multiple sclerosis. Nat. Med. 24 , 1837–1844 (2018).

Hughes, A. N. & Appel, B. Oligodendrocytes express synaptic proteins that modulate myelin sheath formation. Nat. Commun. 10 , 4125 (2019).

Genc, B. et al. Apical dendrite degeneration, a novel cellular pathology for Betz cells in ALS. Sci. Rep. 7 , 41765 (2017).

Wainger, B. J. et al. Effect of ezogabine on cortical and spinal motor neuron excitability in amyotrophic lateral sclerosis: a randomized clinical trial. JAMA Neurol. 78 , 186–196 (2021).

O’Rourke, J. G. et al. C9orf72 is required for proper macrophage and microglial function in mice. Science 351 , 1324–1329 (2016).

McCauley, M. E. et al. C9orf72 in myeloid cells suppresses STING-induced inflammation. Nature 585 , 96–101 (2020).

Ennerfelt, H. et al. SYK coordinates neuroprotective microglial responses in neurodegenerative disease. Cell 185 , 4135–4152 e4122 (2022).

Murphy, A. E., Fancy, N. & Skene, N. Avoiding false discoveries in single-cell RNA-seq by revisiting the first Alzheimer’s disease dataset. eLife https://doi.org/10.7554/eLife.90214 (2023).

Gautier, O. et al. Challenges of profiling motor neuron transcriptomes from human spinal cord. Neuron 111 , 3739–3741 (2023).

Marsh, S. E. et al. Dissection of artifactual and confounding glial signatures by single-cell sequencing of mouse and human brain. Nat. Neurosci. 25 , 306–316 (2022).

Krienen, F. M. et al. Innovations present in the primate interneuron repertoire. Nature 586 , 262–269 (2020).

Mitchell, J. M. et al. Mapping genetic effects on cellular phenotypes with ‘cell villages’. Preprint at bioRxiv https://doi.org/10.1101/2020.06.29.174383 (2020).

Rapino, F. et al. Small-molecule screen reveals pathways that regulate C4 secretion in stem cell-derived astrocytes. Stem Cell Rep. 18 , 237–253 (2023).

Article   CAS   Google Scholar  

Hao, Y. et al. Integrated analysis of multimodal single-cell data. Cell 184 , 3573–3587 e3529 (2021).

Raudvere, U. et al. g:Profiler: a web server for functional enrichment analysis and conversions of gene lists (2019 update). Nucleic Acids Res. 47 , W191–W198 (2019).

Szklarczyk, D. et al. STRING v11: protein–protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets. Nucleic Acids Res. 47 , D607–D613 (2019).

Subramanian, A. et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc. Natl Acad. Sci. USA 102 , 15545–15550 (2005).

Pietilainen, O. et al. Astrocytic cell adhesion genes linked to schizophrenia correlate with synaptic programs in neurons. Cell Rep. 42 , 111988 (2023).

Fukuda, A. et al. De novo DNA methyltransferases DNMT3A and DNMT3B are essential for XIST silencing for erosion of dosage compensation in pluripotent stem cells. Stem Cell Rep. 16 , 2138–2148 (2021).

Hao, J. et al. Loss of TBK1 activity leads to TDP-43 proteinopathy through lysosomal dysfunction in human motor neurons. Preprint at bioRxiv https://doi.org/10.1101/2021.10.11.464011 (2021).

Download references

Acknowledgements

We thank the study participants and staff at Massachusetts Alzheimer’s Disease Research Center for sequencing (ADRC) (NIA P50 AG005134). We thank the study participants and K. Wilsbach, L. Ostrow and staff at Target ALS Neuropathology core and associated institutions for validation of cohort samples. We thank UCB Pharma for partially funding these studies. D.M. acknowledges support from Massachusetts ADRC (5P50AG005134) pilot project and the National Institute of Neurological Disorders and Stroke (NINDS) (K08NS104270). We also like to thank P. Tesar and his group for invaluable discussions on oligodendroglial biology. Some figures were created with BioRender.com under license to the Eggan lab (2020–2022).

Author information

These authors contributed equally: Francesco Limone, Daniel A. Mordes.

Authors and Affiliations

Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA

Francesco Limone, Daniel A. Mordes, Alexander Couto, Brian J. Joseph, Jana M. Mitchell, Sulagna Dia Ghosh, Yingying Zhang, Aaron Burberry, Olli Pietiläinen & Kevin Eggan

Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA, USA

Francesco Limone, Daniel A. Mordes, Jana M. Mitchell, Martine Therrien, Sulagna Dia Ghosh, Daniel Meyer, Yingying Zhang, Melissa Goldman, Laura Bortolin, Beth Stevens, Steven A. McCarroll, Aaron Burberry, Olli Pietiläinen & Kevin Eggan

Neuroscience Institute, NYU Grossman School of Medicine, New York, NY, USA

Francesco Limone

Department of Pathology, Massachusetts General Hospital, Boston, MA, USA

Daniel A. Mordes & Inma Cobos

FM Kirby Neurobiology Center, Boston Children’s Hospital, Boston, MA, USA

Martine Therrien & Beth Stevens

Department of Genetics, Harvard Medical School, Boston, MA, USA

Sulagna Dia Ghosh, Daniel Meyer, Melissa Goldman, Laura Bortolin & Steven A. McCarroll

Howard Hughes Medical Institute, Boston, MA, USA

Beth Stevens

Neuroinflammation Focus Area, UCB Pharma, Braine-l’Alleud, Belgium

Irena Kadiu

Department of Pathology, School of Medicine, Case Western Reserve University, Cleveland, OH, USA

Aaron Burberry

Neuroscience Center, Helsinki Institute of Life Science, University of Helsinki, Helsinki, Finland

Olli Pietiläinen

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Contributions

The study was designed by F.L. and D.A.M. and directed and coordinated by K.E. and S.A.M. with input from B.S. and I.K. Manuscript writing was performed by F.L. and D.A.M., with support from O.P. and A.B. F.L. performed bioinformatics analysis with the help of S.D.G., D.M. and D.A.M. D.A.M. and I.C. supported obtaining postmortem samples and carried out nuclei isolation and RNA sequencing with M.G. and L.B.; Y.Z., F.L., M.T., O.P., A.B., A.C. and B.J.J. performed bioinformatics analyses of bulk RNA sequencing and helped with protein and RNA validation with cellular models; F.L. and J.M.M. performed analysis of published datasets; F.L., M.T. and B.S. contributed to microglial biology section; and K.E. acquired primary funding.

Corresponding authors

Correspondence to Francesco Limone or Kevin Eggan .

Ethics declarations

Competing interests.

I.K. is an employee at UCB Pharma and holds stock options. K.E. is a cofounder of Q-State Biosciences, Quralis, Enclear Therapies and is group vice-president at BioMarin Pharmaceutical. The remaining authors declare no competing interests.

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Extended data

Extended data fig. 1 technical parameters of snrnaseq and cell-type distribution across individuals..

a . Schematic diagram of workflow for isolation of nuclei from cortices of ALS patients and age-matched controls followed by single-cell RNA sequencing by DropSeq, library generation and Quality Controls for analysis with Seurat 3.0.2 b . Frozen tissue from one of the individuals. c . Staining for TDP-43 in one of the patient sample, note neuron with skein-like inclusions and faint nuclear staining (scale bar 25 μ m, n = 3 patients analysed). d . Quality controls post-filtering (FC – Frontal Cortex): number of total nuclei detected (barcodes), average number of genes per nucleus (nFeatures), and average number of UMIs (Unique Molecular Identifiers) per nucleus (nCounts). e . t -SNE projections of the whole cohort with expression of broad cell type markers. f . Dotplot representing percentage of cells expressing additional cell type specific markers. g . t -SNE distribution of whole cohort with annotated cell types split by diagnosis (ALS patients n  = 5, age-matched Controls n  = 3, n  = 79,169 total nuclei). h . Quality controls post-filtering (FC – Frontal Cortex): number of total nuclei detected (barcodes), average number of genes per nucleus (nFeatures), and average number of UMIs (Unique Molecular Identifiers) per nucleus (nCounts). i . Fraction of each cell types identified in whole cohort split by diagnosis (mean ± SEM, ALS patients n  = 5, age-matched Controls n  = 3).

Extended Data Fig. 2 Expression of ALS-FTD associated genes in different cellular subtypes and excitatory neurons subtypes.

a . Dotplot representing expression of gene associated with the ALS-FTD spectrum in each cell type identified in the whole cortex split by diagnosis. b . t -SNE projection of excitatory neurons clusters (ALS n  = 15,227 nuclei, Control n  = 17,583 nuclei). c . t -SNE projection of subclusters identified in excitatory neurons represents different, biologically relevant neuronal layers ( FindNeighbor (res=0.2)). d . Dotplot representing percentage of cells expressing broad markers for different cortical layers. e . Distribution of excitatory neurons in subclusters by diagnosis (mean ± SEM). f . t -SNE projection of excitatory neurons by clusters and by diagnosis. g . Gene Set Enrichment Analysis for the ALS-FTD associated genes in Exc1 excitatory neuron subtype. h . Heatmap representing expression of gene associated with the ALS-FTD spectrum in each excitatory neurons identified split by diagnosis.

Extended Data Fig. 3 L5-ETNs/CSMNs-like neurons express higher levels of ALS-FTD related genes.

a,b , Spotplot and corresponding boxplot from Maynard et al. for the expression of layer Vb Motor Cortex marker, SNCG and THY1 , identified as enriched in Exc1 (boxplots for mean and interquantile ranges + -SD). c,d . Dotplot and representative Violin plots for markers of L5 ExtraTelencephalic neurons of human Motor Cortex in Schirmer et al. e,f . Dotplot and representative Violin plots for markers of L5 ExtraTelencephalic neurons of human Motor Cortex in Velmeshev et al. (geometric boxplots for median and interquantile ranges) g-i . Dotplot representing expression of Layer V markers (d), von Economo markers (e), LR-SCPN markers (f) in Schirmer et al. and Velmeshev et al. l . Violin plots and corresponding Gene Set Enrichment Analysis of z-scores for expression of ALS-FTD-associated genes in THY1-neurons identified by Schimer et al. (bars denote median). m . Violin plots and corresponding Gene Set Enrichment Analysis of z-scores for expression of ALS-FTD-associated genes in THY1-neurons identified by Velmeshev et al. (bars denote median). n . Boxplot from Maynard et al. for the expression of additional top ALS/FTD associated genes identified in Exc1 (geometric boxplots for mean and interquantile ranges + -SD) (n=multiple individuals as reported in publicly available datasets)

Extended Data Fig. 4 Classes of L5-ETNs express higher levels of stress pathways.

a . DGE-based PCA plots showing separation of individuals by diagnosis in Exc1. b . Violin plots of z-scores for genes upregulated in each class of L5-ETNs (DGE1, DGE5, DGE6) in the three groups split by diagnosis (bars denote median – symbols: average score per individual). c , Comparison of DGEs signature (>10% cells, >2-FC, adj.p < 0.05) and redoDGE signatures identified in each subgroup with random seeding of equal cell numbers per diagnosis (>10% cells, >2-FC, adj.p < 0.05, max.cells.per.ident=lowest.number.per.group). d . Violin plots of z-scores for redoDGE signatures (median and interquantile ranges – symbols: average score per individual). e . Correlation plot of Log Fold Changes in ALLminus1 comparisons. f . Venn diagram comparing genes shared between ALLminus1 analyses. g . Venn diagram comparing genes in common between ALLminus1 analysis and ExcAll. h . Violin plots of z-scores for genes globally downregulated in Excall (geometric boxplots represent median and interquantile ranges – symbols: average score per individual). i . Comparison of genes globally upregulated in ALS (DGEall) with genes upregulated in CUX2-exc0 (>10% of cells, >2-FC, adj.p < 0.05) j . Gene Ontology analysis of terms for DGE0 highlighted terms involved in synaptic biology (CC=Cellular Components). k. Gene Ontology analysis of terms for DGEall highlighted terms involved in synaptic biology (CC=Cellular Components). l , m. Selected GO terms for DGE0, DGE1 and DGEall. n. Venn Diagram for shared upregulated genes between other excitatory neurons and global signature. o , p. Venn Diagrams for shared upregulated genes between treated MS excitatory neurons from Schirmer et al. and AD excitatory neurons from Mathys et al. with changes found in this study.

Extended Data Fig. 5 Global protein-protein interaction network for genes upregulated in ALS excitatory neurons.

Color-coding derived from MCL clustering to identified closely related groups of proteins.

Extended Data Fig. 6 Proteostatic stress in hPSC-derived neurons resembles changes in excitatory neurons from brain of ALS patients.

a . Diagram of neuronal differentiation from Pluripotent Stem Cells and treatment with proteasome inhibitors for bulk RNA-sequencing. b . Quantification of proteasome inhibition (mean ± SD, n = 3 biological replicates, n = 9 technical total, (t-test, ****p < 0.001). c . Principle Component Analysis plot showing strong effect of treatments compared to untreated controls (n = 3 biological replicates). d . Protein-protein interaction network of shared genes. e . Venn Diagram depicting shared upregulated genes in treated hPSC-derived neurons (MG132), excitatory neurons from ALS patients (DGEall) and genes misregulated in human neurons after TDP-43 siRNA from Klim et al. (proportional test) f-g . Western Blot quantification of soluble and insoluble TDP-43 from motor cortex of ALS patients and age-matched controls (t-test).

Extended Data Fig. 7 Oligodendrocytes polarize between myelinating and neuro-engaged states.

a,b . t -SNE projection and Violin plot of markers of Oligodendrocyte Progenitor Cells (OPCs) and mature oligodendrocytes (geometric boxplots represent median and interquantile ranges – symbols: log2(AverageExpression) per individual) (fraction of cell expressing). c,d . t -SNE projection of markers of actively myelinating oligodendrocytes and violin plots representing z-score for selected GO terms by cluster (geometric boxplots represent median and interquantile ranges – symbols: average score per individual). e,f . t -SNE projection of markers of neuronally-engaged oligodendrocytes and violin plots representing z-score for selected GO terms by cluster (geometric boxplots represent median and interquantile ranges – symbols: average score per individual). g . DGE-based PCA plot showing separation of individuals by diagnosis. h . Violin plot for z-score for genes upregulated and downregulated in ALS patients (geometric boxplots represent median and interquantile ranges – symbols: average score per individual). i . Correlation plot of Log Fold Changes in ALLminus1 comparisons. j . Venn diagram comparing genes shared between ALLminus1 analyses. k . Venn diagram comparing genes in common between ALLminus1 analysis and OligoAll. l . Violin plot of markers of actively myelinating oligodendrocytes (geometric boxplots represent median and interquantile ranges – symbols: log2(AverageExpression) per individual) (fraction of cell expressing). m . Violin plot of markers of neuronally-engaged oligodendrocytes (geometric boxplots represent median and interquantile ranges – symbols: log2(AverageExpression) per individual) (fraction of cell expressing). n , o . GO analysis for genes downregulated and upregulated in ALS oligodendrocytes, highlighted terms involved in myelination (CC=Cellular Component). p . Dotplot representing genes characteristic of maturation and development of OPCs in myelinating oligodendrocytes in each subcluster split by diagnosis.

Extended Data Fig. 8 Comparison of ALS-driven changes with study with similar signature disrupted in disease (MS).

a,b . t -SNE projection and violin plot representing z-score for highly myelinating, OPALIN + oligodendrocytes in Jäkel et al (geometric boxplots represent median and interquantile ranges – symbols: average per individual). c . Comparison of genes downregulated in oligodendroglia from ALS patients with genes characteristic of highly myelinating, OPALIN + subtypes identified by this study (oliglia0) and by Jäkel et al (Jäkel6), highlighted genes are shared with GO terms shown in figures. d,e . t -SNE projection and violin plot representing z-score for genes of mature, not-actively myelinating oligodendrocytes in Jäkel et al (geometric boxplots represent median and interquantile ranges – symbols: average per individual). f,g . Comparison of genes upregulated in oligodendroglia from ALS patients with genes characteristic of mature, lowly myelinating groups in this study (oliglia1 and 4) and by Jäkel et al (Jäkel1), highlighted genes are shared with GO terms shown in figures. h . Dotplot representing z-scores for the genetic signatures identified in the actively myelinating cells, the mature lowly myelinating cells and DEGs identified in this study. (for whole panel n  =  3 Control individuals, n  =  5 sALS patients)

Extended Data Fig. 9 Shared features between ALS-driven changes and reactive subcluster of microglia.

a . Correlation plot of Log Fold Changes in ALLminus1 comparisons. b . Venn diagram comparing genes shared between ALLminus1 analyses. c . Venn diagram comparing genes in common between ALLminus1 analysis and MicroAll. d . DGE-based PCA plot showing separation of individuals by diagnosis. e . Violin plot for z-score for genes upregulated and downregulated in ALS patients (geometric boxplots represent median and interquantile ranges – symbols: average score per individual). f . t-SNE projection of subclusters identified within microglia (Micro0 = Homeo = homeostatic, Micro1 = DAMs = Disease-associated microglia, Micro2 = Cycling cells)). g . Distribution of microglia within clusters by diagnosis. h . Dotplot representing genes identified as characteristic of Homeostatic microglia and DAMs by subcluster. i . Dotplot representing genes identified as characteristic of Homeostatic microglia and DAMs by diagnosis. j . Volcano plot of statistically significant differentially expressed genes between Control and ALS microglia (top ten upregulated and top ten downregulated genes highlighted). k . Violin plots of representative DEGs downregulated in ALS patients of genes associated with homeostatic microglia (geometric boxplots represent median and interquantile ranges – symbols: log2(AverageExpression) per individual) (fraction of cell expressing). l . Gene Ontology analysis of terms associated with genes characteristic of DAMs microglia, highlighted terms playing important role in microglial biology and/or pathogenesis of the disease. m . t-SNE projections representing z-score for selected, statistically significant GO terms.

Extended Data Fig. 10 Apoptotic neurons upregulate lysosomal genes in microglia.

a . Schematic of workflow and results from the Connectivity Map project for the genes upregulated in ALS microglia. Heatmap shows what cellular signature is most closely related to the query. b . Diagram of microglia and neuronal differentiation from Pluripotent Stem Cells, induction of apoptosis neurons and feeding to iMGLs. c . RT-qPCR quantification of cell cycle-associated genes after feeding (t-test, *p < 0.05, **p < 0.01, ***p < 0.001; n = 3 biological replicates).

Supplementary information

Reporting summary, supplementary data 1.

Graphical abstract and working model. Our study highlights cell type-specific changes in premotor/motor cortex of patients with sporadic ALS. Specifically, we identify upregulation of synaptic molecules in excitatory neurons of upper cortical layers, interestingly correlating to hyperexcitability phenotypes seen in patients. Moreover, excitatory neurons of the deeper layers of the cortex, which project to the spinal cord and are most affected by the disease, show higher levels of cellular stresses than other neuronal types. Correspondently, oligodendrocytes transition from a highly myelinating state to a more neuronally engaged state, probably to counteract stressed phenotypes seen in excitatory neurons. At the same time, microglia show a reactive state with specific upregulation of endolysosomal pathways.

Source Data

Unmodified western blots from all manuscripts.

Source data tables: (1) Patient information. (2) Disease-associated genes used for module scores. (3) Genes upregulated in each neuronal subtype in patients with ALS. (4) Genes used for oligodendrocytes module scores.

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Limone, F., Mordes, D.A., Couto, A. et al. Single-nucleus sequencing reveals enriched expression of genetic risk factors in extratelencephalic neurons sensitive to degeneration in ALS. Nat Aging (2024). https://doi.org/10.1038/s43587-024-00640-0

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DOI : https://doi.org/10.1038/s43587-024-00640-0

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Drug Combination Slows Progression Of ALS And Could Mark 'New Era' In Treatment

Jon Hamilton

Patients with a fast-progressing form of ALS who got daily doses of an experimental two-drug combination called AMX0035 scored higher on a standard measure of function than patients who didn't get the drug. Zephyr/Science Source hide caption

Patients with a fast-progressing form of ALS who got daily doses of an experimental two-drug combination called AMX0035 scored higher on a standard measure of function than patients who didn't get the drug.

A combination of two experimental drugs appears to slow the decline of patients with amyotrophic lateral sclerosis, an illness often known as ALS or Lou Gehrig's disease.

A six-month study of 137 patients with a fast-progressing form of the disease found that those who got daily doses of a two-drug combination called AMX0035 scored several points higher on a standard measure of function, a team reports in the Sept. 3 issue of The New England Journal of Medicine .

The difference was modest but meaningful to patients, said Dr. Sabrina Paganoni . She's the lead author and a researcher at the Sean Healey & AMG Center for ALS at Mass General and Harvard Medical School.

"They want to be able to continue to use their hands so they can cut their own food and type emails, or they want to be able to walk and climb stairs," Paganoni said. "And this is exactly what we measured in the trial."

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Heads Still Dry, Scientists Try New Approach With ALS

The results are far from a cure. Even so, "I am convinced that we are at the beginning of a new era in ALS treatment discovery," Paganoni said.

"There's great hope for disease-modifying treatment," added Tania Gendron , who studies neurodegenerative diseases at the Mayo Clinic in Jacksonville and was not involved in the study. "In the next few years I think there are going to be some big discoveries."

ALS destroys the nerve cells that control muscle movement. Patients typically become disabled and die within five years of their diagnosis.

For decades, the only drug approved by the Food and Drug Administration for ALS was riluzole , which has been on the market since 1995 and has been shown to extend the lives of patients. Then in 2017, the FDA approved edaravone , which helps some patients retain function longer.

AMX0035 works by protecting nerve cells from two types of damage that are hallmarks of ALS. And in the study it produced a benefit, even though many of the patients were already taking riluzole and edaravone.

It appears that the new and old drugs all work in different ways to slow down the disease, Paganoni said. "We think ultimately we will need a combination of treatments to effectively fight ALS."

It's not clear yet whether AMX0035 extends life or maintains muscle strength. And ordinarily at least one larger study would be required before the FDA considered approving the drug.

But the ALS Association along with the advocacy group I AM ALS have joined forces to petition the FDA to make an exception.

"In ALS, a trial like this would probably take about three years," said Neil Thakur, chief mission officer of the ALS Association. "And so the question for the whole community is what do we gain for that three-year study?"

The ALS Association helped fund the research on AMX0035 and has a limited financial stake in its success. The group's main concern, though, is patients who won't live long enough to wait for another study, Thakur said.

"That's why we're thinking the best thing to do for the community is to make this drug available sooner and let everyone have it as a treatment option as soon as possible," he said.

AMX0035 has taken a highly unusual path toward approval.

It was developed by Amylyx , a tiny company founded by a couple of college students, Josh Cohen and Justin Klee, who are still in their 20s. The pair were working late one night in the company's Cambridge, Mass., office when they learned the results of the ALS study.

"When the statisticians called, you could hear their whole firm cheering in the background," Cohen said. "So we knew before they said the numbers that something good had happened."

But their elation was mixed with a sense of responsibility, Klee said.

"While these results are great, it's not a cure, and so we and others in the whole community need to keep pushing forward until we get cures," he said.

That's beginning to look more likely than it did even a few years ago, the Mayo Clinic's Gendron said.

One reason is that scientists are discovering new biological markers that appear in the blood or spinal fluid of ALS patients. These markers will allow earlier diagnosis when treatments are more likely to work, Gendron said. They will also help select which patients to give a particular drug and allow researchers to know whether the drug is reaching its intended target.

Another reason for optimism is the sheer amount of research going on.

"What makes this time so exciting is there are over 50 different clinical trials that are enrolling and recruiting ALS patients right now," said Kuldip Dave, the ALS Association's vice president of research. "And they're all going after different targets."

• Brain research
• neuroscience
• Lou Gehrig's Disease

Amyotrophic Lateral Sclerosis (ALS)

What is amyotrophic lateral sclerosis (als).

Amyotrophic lateral sclerosis (ALS), formerly known as Lou Gehrig's disease, is a neurological disorder that affects motor neurons, the nerve cells in the brain and spinal cord that control voluntary muscle movement and breathing. As motor neurons degenerate and die, they stop sending messages to the muscles, which causes the muscles to weaken, start to twitch (fasciculations), and waste away (atrophy). Eventually, in people with ALS, the brain loses its ability to initiate and control voluntary movements such as walking, talking, chewing and other functions, as well as breathing. ALS is progressive, meaning the symptoms get worse over time.

The U.S. Food and Drug Administration has approved several drugs for ALS that may prolong survival, reduce the rate of decline, or help manage symptoms. However, there is currently no known treatment that stops or reverses the progression of ALS.

Early symptoms include:

• Muscle twitches in the arm, leg, shoulder, or tongue
• Muscle cramps
• Tight and stiff muscles (spasticity)
• Muscle weakness affecting an arm, a leg, or the neck
• Slurred and nasal speech
• Difficulty chewing or swallowing

As the disease progresses, muscle weakness and atrophy spread to other parts of your body. People with ALS may develop problems with:

• Chewing food and swallowing (dysphagia)
• Drooling (sialorrhea)
• Speaking or forming words (dysarthria)
• Breathing (dyspnea)
• Unintended crying, laughing, or other emotional displays (pseudobulbar symptoms)
• Constipation
• Maintaining weight and getting enough nutrients

Eventually, people with ALS will not be able to stand or walk, get in or out of bed on their own, use their hands and arms, or breathe on their own. Because they usually remain able to reason, remember, and understand, they are aware of their progressive loss of function. This can cause anxiety and depression in the person with ALS and their loved ones. Although not as common, people with ALS also may experience problems with language or decision-making. Some also develop a form of dementia known as FTD-ALS .

Most people with ALS die from being unable to breathe on their own (known as respiratory failure,) usually within three to five years from when the symptoms first appear. However, about 10% survive for a decade or more.

Who is more likely to get amyotrophic lateral sclerosis (ALS)?

A risk factor is a condition or behavior that occurs more frequently in those who have a disease, or who are at greater risk of getting a disease, than in those who don't have the risk factor. Having a risk factor doesn't mean a person will develop a disorder, and not having a risk factor doesn't mean you won’t. Risk factors for ALS include:

• Age —Although the disease can strike at any age, symptoms most commonly develop between the ages of 55 and 75.
• Biological sex —Men are slightly more likely to develop ALS than women. However, at older ages, men and women are equally likely to be diagnosed with ALS.
• Race and ethnicity —Whites and non-Hispanics are most likely to develop the disease, but ALS affects people of all races and ethnic backgrounds.

Some studies suggest military veterans are about one and a half to two times more likely to develop ALS, although the reason for this is unclear. Possible risk factors for veterans include exposure to lead, pesticides, and other environmental toxins. Some studies have also shown that head injury can be associated with higher risk for ALS, but more research is needed to understand this connection.

Sporadic and Familial ALS

Nearly all cases of ALS are considered sporadic, meaning the disease seems to occur at random with no clearly associated risk factors and no family history of the disease. Although family members of people with ALS are at an increased risk for the disease, the overall risk is very low, and most will not develop ALS.

About 10% of all ALS cases are familial (also called inherited or genetic). Changes in more than a dozen genes have been found to cause familial ALS.

• About 25-40% of all familial cases (and a small percentage of sporadic cases) are caused by a defect in the C9orf72 gene. C9orf72 makes a protein found in motor neurons and nerve cells in the brain.
• Another 12-20% of familial cases result from mutations in the SOD1 gene. SOD1 is involved in production of the enzyme copper-zinc superoxide dismutase 1.

In 2021, a team of scientists led by the NIH and the Uniformed Services University of the Health Sciences announced it had discovered a unique form of genetic ALS that affects children as early as age 4 years . This childhood form is linked to the gene SPTLC1 that is part of the body's fat production system and may be caused by changes in the way the body metabolizes fatty materials (lipids).

How is amyotrophic lateral sclerosis (ALS) diagnosed and treated?

Diagnosing als.

It is important to get an accurate ALS diagnosis as soon as possible. ALS treatments may be most effective early in the course of the disease. A neurologist familiar with ALS can help a person get diagnosed early after symptom onset.

There is no single test that can definitely diagnose ALS. A healthcare provider will conduct a physical exam and review the person’s full medical history. A neurologic examination will test reflexes, muscle strength, and other responses. These tests should be performed at regular intervals to assess whether symptoms are getting worse over time.

A healthcare provider may conduct muscle and imaging tests to rule out other diseases. This can help support an ALS diagnosis. These tests include:

• A nerve conduction study (NCS) — measures the electrical activity of nerves and muscles by assessing the nerve's ability to send a signal along the nerve or to the muscle.
• A needle exam — a recording technique that detects electrical activity in muscle fibers using a needle electrode.
• Magnetic resonance imaging (MRI) — uses a magnetic field and radio waves to produce detailed images of the brain and spinal cord.

Blood and urine tests may be performed based on the person’s symptoms, test results, and findings from a neurological exam. In some cases, a spinal tap (lumbar puncture) may be performed to obtain the fluid that surrounds the brain and spinal cord called cerebrospinal fluid (CSF) for additional testing. A physician may order these tests to eliminate the possibility of other diseases. A muscle biopsy may be performed to help determine whether the person may have a muscle disease other than ALS.

Treating ALS

There is no treatment to reverse damage to motor neurons or cure ALS at this time. However, some treatments may slow progression of the disease, improve quality of life, and extend survival. New treatments have become available in the past several years, and researchers continue to explore diverse avenues to slow or stop progression of ALS.

Supportive health care is best provided by integrated, multi-disciplinary teams of professionals that may include physicians, pharmacists, physical, occupational, speech, and respiratory therapists, nutritionists, social workers, clinical psychologists, and home care and hospice nurses. These teams can design an individualized treatment plan and provide special equipment aimed at keeping people as mobile, comfortable, and independent as possible.

Doctors may use the following medications approved by the U.S. Food and Drug Administration (FDA) to support a treatment plan for ALS:

• Riluzole (Rilutek) is an oral medication believed to reduce damage to motor neurons by decreasing levels of glutamate, which transports messages between nerve cells and motor neurons. Clinical trials in people with ALS showed that riluzole may prolong survival by a few months. The thickened liquid form (Tiglutik) or the tablet (Exservan) that dissolves on the tongue may be preferred if the person has swallowing difficulties.
• Edaravone (Radicava) is an antioxidant given either orally or intravenously and has been shown to slow functional decline in some people with ALS. RADICAVA ORS is a form of edaravone that can be taken orally or via feeding tube.
• Sodium phenylbutyrate/taurursodiol (Relyvrio) is an oral medication that was proposed to prevent nerve cell death by blocking stress signals in cells.  The FDA approved Relyvrio based on safety and efficacy data from a single, smaller ALS clinical trial in September 2022. However, a larger clinical trial failed to confirm the earlier findings, and the manufacturer of Relyvrio removed the drug from the market in 2024.
• Tofersen (Qalsody) is given through a spinal injection to people with ALS who have been determined to have a mutation in the SOD1 gene. While the benefits of this drug are still under study, it may work by decreasing one of the markers of damage to neurons.

A doctor may prescribe other medications or treatments to help manage symptoms, including muscle cramps and stiffness, excessive saliva and phlegm, and unwanted episodes of crying and/or laughing, or other emotional displays. Medications may also help with any pain, depression, sleep disturbances, or constipation.

Rehabilitation, therapy, and other support

A treatment plan for ALS usually includes rehabilitation, which should be tailored to the person’s individual needs and may include physical, occupational, and speech therapy.

Support for physical function and daily life

Physical therapy can help the person with ALS maintain function, including lowering their risk of falls and joint pain and maximizing their independence at different stages of the disease. Low-impact exercises such as walking, swimming, or using a stationary exercise bike along with range of motion exercises can help maintain muscle strength and function. Occupational therapists can help with activities of daily living and self-care. They can also suggest assistive devices for feeding, bathing, and grooming so that the person can be as independent as possible.

Speech and communication support

Speech therapists can help people with ALS learn strategies to speak louder and more clearly and help maintain the ability to communicate. Computer-based speech synthesizers use eye-tracking devices that allow a person to navigate the web and to type on custom screens to communicate. Voice banking is a process sometimes used by people with ALS to store their own voice for future use in computer-based speech synthesizers.

A brain-computer interface (BCI) is a system that allows individuals to communicate or control equipment such as a wheelchair using only brain activity. Researchers are developing more efficient, mobile BCIs for people with severe paralysis and/or visual impairments.

Support for nutrition, breathing, and feeding

People with ALS may have trouble chewing and swallowing their food, and getting the nutrients they need. Nutritionists and registered dieticians can help plan small, nutritious meals throughout the day and identify foods to avoid. When the person can no longer eat with help, a feeding tube can reduce the person’s risk of choking and pneumonia.

As the muscles responsible for breathing start to weaken, individuals with ALS may have shortness of breath during physical activity and difficulty breathing at night or when lying down. Noninvasive ventilation (NIV) is a type of breathing support that is usually delivered through a mask over the nose and/or mouth. It may help decrease the discomfort of breathing in some individuals with ALS. Initially, NIV may only be necessary at night, but may eventually be used full time. As the disease progresses, the person may need the support of respirators (mechanical ventilators) to inflate and deflate the lungs.

Because the muscles that control breathing become weak, people with ALS also may have trouble generating a strong cough. There are several techniques to increase forceful coughing, including mechanical cough assistive devices.

Caring for a person living with ALS

As the person with ALS progresses in their disease, they will need more and more help with daily activities. Being a caregiver for a person with ALS, while rewarding, can be challenging for the person’s loved ones and caregivers. It is important for caregivers take care of themselves and to seek support when needed. Free and paid resources are available to provide home health care services and support. Visit the organizations listed at the end of this article to find support in your area.

What are the latest updates on amyotrophic lateral sclerosis (ALS)?

NINDS is the primary federal funder of research on the brain and nervous system, including disorders such as ALS. NINDS is a component of the NIH, the leading supporter of biomedical research in the world. In 2023, NINDS published strategic priorities for ALS research (pdf, 1818 KB) to accelerate the development of effective interventions for the diagnosis, treatment, management, prevention, or cure of ALS. These strategic priorities were developed with input from scientists, clinicians, advocates, people affected by ALS, and the public. Under the Accelerating Access to Critical Therapies for ALS Act , NINDS also funds research on expanded access for investigational new drugs to people living with ALS who are not eligible for clinical trials.

Scientific discoveries have resulted in the identification of multiple therapeutic targets for ALS, and four disease-modifying, plus one symptom-managing, ALS therapies have been approved by the FDA. However, the impact of these disease-modifying therapies is modest. To develop truly effective ALS treatments, we must address numerous challenges. The goals of NINDS’s ALS research are to understand the cellular mechanisms involved in the development and progression of the disease, investigate the influence of genetics and other potential risk factors, identify biomarkers, and develop new treatments.

Cellular defects

Ongoing studies seek to understand the mechanisms that selectively trigger motor neurons to degenerate in ALS, which may lead to effective approaches to stop this process. Research using cellular culture systems and animal models suggests that motor neuron death is caused by a variety of cellular defects, including those involved in protein recycling and gene regulation, as well as structural impairments of motor neurons. Increasing evidence also suggests that glial support cells and inflammation cells of the nervous system may play an important role in ALS.

Scientists are turning adult skin and blood cells into stem cells that are capable of becoming any cell type, including motor neurons and other cells which may be involved in ALS. NINDS-funded scientists are using stem cells to grow human spinal cord sections on tissue chips to help better understand the function of neurons involved in ALS.

Genetics and epigenetics

Clinical research studies supported by NINDS are looking into how ALS symptoms change over time in people with C9orf72 mutations. Other studies are working to identify additional genes that may cause or put a person at risk for either familial or sporadic ALS.

A large-scale collaborative research effort supported by NINDS, other NIH institutes, and several public and private organizations is analyzing genetic data from thousands of individuals with ALS to discover new genes involved in the disease. By using novel gene sequencing tools, researchers are now able to rapidly identify new genes in the human genome involved in ALS and other neurodegenerative diseases. People who carry genes associated with ALS may be able to participate in long-term, observational studies to help researchers understand how the disease progresses over time in diverse populations.

Additionally, researchers are looking at the potential role of epigenetics in ALS development. Epigenetic changes can switch genes on and off during a person’s lifetime, which can greatly impact both health and disease. Although this research is exploratory, scientists hope that understanding epigenetics can offer new information about how ALS develops.

Biomarkers  

NINDS supports research on the development of biomarkers—biological measures of a disease. Biomarkers can be molecules derived from a bodily fluid (blood or cerebrospinal fluid), an image of the brain or spinal cord, or a measure of the ability of a nerve or muscle to process electrical signals. ALS biomarkers can help identify the rate of progression and the effectiveness of current and future therapies.

For research articles and summaries on ALS, search  PubMed , which contains citations from medical journals and other sites.

How can I or my loved one help improve care for people with amyotrophic lateral sclerosis (ALS)?

The  National ALS Registry  collects, manages, and analyzes de-identified data about people with ALS in the United States. Developed by the Center for Disease Control and Prevention's Agency for Toxic Substances and Disease Registry (ATSDR), this registry establishes information about the number of ALS cases, collects demographic, occupational, and environmental exposure data from people with ALS to learn about potential risk factors for the disease, and notifies participants about research opportunities. The Registry includes data from national databases as well as de-identified information provided by individuals with ALS. All information is kept confidential. People with ALS can add their information to the registry and sign up to receive for more information.

Consider participating in a clinical trial so clinicians and scientists can learn more about ALS. Clinical research uses human study participants to help researchers learn more about a disorder and perhaps find better ways to safely detect, treat, or prevent disease.

All types of study participants are needed— those who are healthy or may have an illness or disease— of all different ages, sexes, races, and ethnicities to ensure that study results apply to as many people as possible, and that treatments will be safe and effective for everyone who will use them.

For information about participating in clinical research visit  NIH Clinical Research Trials and You . Learn about clinical trials currently looking for people with ALS at  Clinicaltrial.gov .

NINDS also supports the  NIH NeuroBioBank , a collaborative effort involving several brain banks across the U.S. that supply investigators with tissue from people with neurological and other disorders. Tissue from individuals with ALS is needed to help advance critical research on the disease. A single donated brain can make a huge impact on ALS research, potentially providing information for hundreds of studies. The goal is to increase the availability of, and access to, high quality specimens for research to understand the neurological basis of the disease. Prospective donors can begin the enrollment process by visiting  Learn How to Become a Brain Donor .

Where can I find more information about amyotrophic lateral sclerosis (ALS)? The following organizations and resources help individuals, families, friends, and caregivers of people living with ALS:  ALS Therapy Development Institute Phone: 617-441-7200 Genetic and Rare Diseases (GARD) Information Center Eldercare Locator Phone: 800-677-1116 I AM ALS Phone: 866-942-6257 Les Turner ALS Foundation Phone: 847-679-3311 MedlinePlus Muscular Dystrophy Association Phone: 800-572-1717 National ALS Registry Project ALS Phone: 212-420-7382 The ALS Association Phone: 800-782-4747

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Congress called for an ALS moonshot. The plan for it doesn’t leave Earth

By Bernard Zipprich June 30, 2024

T he National Academies of Sciences, Engineering, and Medicine recently released a congressionally mandated report on how to make amyotrophic lateral sclerosis — a brutal, always fatal condition — a “livable” disease in the next 10 years. Essentially, the committee was tasked with delivering a plan for a moonshot.

As a health care innovation expert and someone living with ALS, I have to say that the report barely gets off the ground.

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ALS is a devastating neurological condition that progressively robs people of their ability to move, speak, eat, and eventually breathe unassisted. Although it’s about as common as multiple sclerosis, because most people with it die within two to five years of diagnosis , only about 30,000 Americans are living with the disease at any one time . There are no effective treatments for 99% of them, and clinical trials keep failing at rates far higher than normal.

To change this bleak outlook, in 2022, Congress commissioned a study and allocated $1 million to identify ways to transform ALS from a fatal condition to a manageable, chronic one “within a decade.” The National Academies of Sciences, Engineering, and Medicine accepted the charge and, to its credit, began studying the issue from a variety of angles. The final report reflects the care that went into the investigation, and includes plenty of good suggestions, such as calls to improve access to high-quality care, eliminate insurance barriers, expand genetic testing, and enhance caregiver support.

Related: Watch: How Harvard researchers boosted an ALS patient’s independence with a box and a balloon

But when it comes to the most important problem of all — accelerating treatment development — the report largely punts.

As someone who provided feedback on a draft of the report, I had high hopes that the final report would highlight promising advances at the scientific frontier and recommend a comprehensive R&D strategy to accelerate treatment development. But the final report does no such thing. Instead, it eschews cutting-edge science in favor of tepid recommendations to do more of the same, like develop better biomarkers and run a new natural history study.

It’s hard to disagree with those generic recommendations. But this isn’t a strategy to end deaths from ALS by 2035.

What might a better strategy entail?

As I suggested to the committee, a comprehensive strategy would accept that ALS, like cancer, is a highly heterogeneous disease. Two cases are rarely alike. Think of it almost like a tree: At the roots, more than 50 different genes can independently trigger 15% to 20% of cases . The rest, while seemingly occurring at random, are likely triggered by a range of environmental, lifestyle, and as-yet-unknown genetic factors. At the trunk, despite this multitude of causal pathways, most cases converge on the breakdown of a protein called transactive response DNA binding protein of 43 kDa (TDP43). At the branches, because TDP43 plays an important role in transcribing DNA into RNA and proteins, genetics and epigenetics likely contribute to important differences in terms of how ALS plays out. This means that drugs that work for one person may not work for another — which is exactly what the research suggests.

This biological context might then inform a three-pronged strategy:

Prong 1 would focus on the trunk of the tree — accelerating the development of drugs focused on addressing TDP43 malfunctions and their proximate effects. Specific recommendations here might include funding to parallel process the preclinical tests necessary to move promising drugs into human trials faster, expanding the size of those trials to enable accelerated approval if warranted by the results, and fast-tracking the development of TDP43 biomarkers and tracers so it’s possible to tell how well such drugs work using medical imaging. (This would also be useful in earlier diagnosis).

Related: Amylyx ALS drug failure leaves patients, advocates, and researchers reeling — and wondering what’s next

Prong 2 would focus on the roots and branches — better defining ALS subtypes to enable precision therapies. Here, I’d want to see expanded efforts to develop gene therapies for the most common genetic forms of ALS and to better understand possible epigenetic subtypes of non-genetic forms. In addition, there should also be a big push to develop disease models that better reflect the heterogeneous nature of ALS. Imagine turning a vial of a patient’s blood into ALS neurons, then using these to identify subtype-specific mechanisms and biomarkers to discover targeted drugs, to run better trials, and eventually to tailor treatments for that individual. This may sound like science fiction, but proofs of concept already exist.

Because breakthrough therapies are still a few years away at best, Prong 3 would focus on R&D initiatives designed to rejuvenate the tree — things like accelerating the availability of brain-computer interfaces that facilitate conversation-speed speech , and more basic research on nerve regeneration .

Some might say that many of these technologies are too nascent and unproven. But that is precisely why they warrant additional focus and investment. Others might say that the idea of targeting treatments at ALS subtypes isn’t yet justified by the science. But it’s known from other heterogeneous conditions, such as lung cancer and MS, that subtype-specific therapies are usually the most transformative. Indeed, the most successful ALS drug so far, tofersen , is a precision therapy for 1% of individuals with a rare genetic ALS subtype. And besides, this is why the strategy proposed here also calls for prioritizing the “trunk.” Maybe we’ll get lucky.

What won’t be achieved if the National Academies’ recommendations for ALS research priorities are accepted as the last word, however, is something different from the same old same old, at least not in the coming decade. To make ALS livable, genuine innovations are needed in the approach to drug development and collaboration between the National Institutes of Health, the Advanced Research Projects Agency for Health, ALS nonprofits, and industry players to make it happen.

The National Academies passed the buck. Who accepts the mantle of leadership is now to be seen.

Bernie Zipprich, M.B.A., is a health care innovation expert and founder of Zipprich Ventures LLC, an advisory practice focused on accelerating treatments for ALS and similar conditions. He is also a trustee of the ALS Association and a research ambassador for the Northeast ALS Consortium. The views expressed here are his own.

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Staying fit may decrease men's risk of ALS

The search for a cure for ALS has been elusive, but researchers may have identified a way to lower a man's risk in the long run.

Staying fit and getting moderate levels of exercise may lower the chances for amyotrophic lateral sclerosis in later life, Norwegian researchers reported Wednesday in the journal Neurology . Advertisement

They did not find a similar link between physical activity and women's risk of ALS, also known as Lou Gehrig 's disease.

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ALS is a neurodegenerative disease that affects nerve cells in the brain and spinal cord. Over time, people lose their ability to eat, speak, move and even breathe. There is no cure.

The new study included more than 373,000 people in Norway (average age, 41). During a follow up that averaged 27 years, 504 developed ALS. Of those, 59% were men. Advertisement

For the study, participants completed a questionnaire about their physical activity level. They listed their activity in one of four categories: sedentary; at least four hours a week of walking or cycling; at least four hours a week of heavy gardening or recreational sports; or participation several times a week in hard training or competitive sports.

So few of the participants put themselves in the most active group that researchers combined the top two categories into one "high-activity" group.

Of the close to 42,000 men who described themselves as most active, 63 developed ALS during the study. Of 77,000 participants with intermediate activity levels, 131 developed ALS, as did 68 of the 29,500 who were least active.

After accounting for factors such as smoking and body mass index, researchers found the most active group had a 41% lower risk than the least active group. Those with moderate levels of activity had a 29% lower risk.

So, the findings show that "not only do moderate to high levels of physical activity and fitness not increase the risk of ALS, but that it may be protective against the disease," Myhre Vaage said in a journal news release. "Future studies of the connection between ALS and exercise are needed to consider sex differences and higher, or professional athlete, activity levels." Advertisement

Researchers noted that men in the lowest of four categories for resting heart rate -- a benchmark of being physically fit -- had a 32% lower risk of ALS compared to men with higher rates.

One limitation of the study was that the activity questionnaire was completed only once. As such, it may not have captured participants' exercise levels over time.

More information

The ALS Association has more about amyotrophic lateral sclerosis .

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Breaking news, exercise may lower the als risk for men — but not women: new study.

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Moderate or vigorous exercise may lower the risk of ALS, a fatal disease, for men but not women, new research finds .

The study, published Wednesday in Neurology , the medical journal of the  American Academy of Neurology , followed 373,696 Norwegian people for about 27 years.

Participants recorded their physical activity, ranging from sedentary to moderate to intense. During the follow-up period, 504 people developed ALS.

Adjusting for lifestyle factors that can affect the risk of ALS, like smoking and body weight, researchers found that male participants who reported moderate levels of physical activity had a 29% lower risk of ALS while high levels of physical activity meant a 41% lower risk.

The study only found an association between physical activity and the risk of ALS in male, not female participants.

Researchers also analyzed participants’ resting heart rates, an indicator of overall fitness, finding that those with the lowest rates had a 32% reduced risk of ALS compared to participants with higher rates.

Study author Dr. Anders Myhre Vaage, of Akershus University Hospital in Norway, notes that the diagnosis of ALS in high-profile athletes has spurred the thinking that strenuous physical activity is an environmental risk factor that leads to the development and early onset of the disease.

One study found that NFL players are four times more likely to develop and die from ALS than the general adult male population.

Research has also shown that ALS risk genes are activated by exercise, adding to the growing debate about the relationship between physical activity and ALS.

“There have been conflicting findings on levels of physical activity, fitness and ALS risk,” Myhre Vaage said. “Our study found that for men, living a more active lifestyle could be linked to a reduced risk of ALS more than 30 years later.”

What is ALS?

ALS, also referred to as Lou Gehrig’s disease, for the Hall of Fame baseball player who died of it in 1941, is a progressive neurodegenerative disease. With ALS, the nerve cells that control muscle function deteriorate, and patients gradually become unable to walk, move, eat, speak and breathe, leading to partial or total paralysis and death.

There is no known cure for ALS — the average life expectancy after diagnosis is two to five years.

Myhre Vaage hopes the study’s findings lead to more research on ALS risk factors.

“Our findings show that, for men, not only do moderate to high levels of physical activity and fitness not increase the risk of ALS, but that they may be protective against the disease,” he said. “Future studies of the connection between ALS and exercise are needed to consider sex differences and higher or professional athlete physical activity levels.”

Other research suggests that the type of physical activity is an important factor in mitigating ALS risk. For example, one study proposed that golfing and gardening put men at three times greater risk of developing ALS.

That study found that golfers and gardeners are especially prone because of frequent exposure to pesticides, which  prior research has tied  to the disease’s development.

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Famous people who have been diagnosed with ALS

According to the ALS Association , someone dies from ALS and someone is diagnosed with it every 90 minutes — and celebrities are not immune to its punishing effects.

Footballer Dwight Clark and physicist, cosmologist and author Stephen Hawking have lost their battles with the disease, while singer Roberta Flack , sportswriter Sarah Langs  and former Chicago Bears football player Steve McMichael are still fighting.

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