Projects Funded 2019

ALS Canada Project Grants

How do unique protein interactions explain TDP-43 behaviour in different people with ALS?

Elucidating the landscape of pathogenic macromolecular assemblies in the motor neurons of ALS patients with TARDBP mutation

$100,000 awarded to Dr. Mohan Babu at the University of Regina

Proteins are large molecules that do most of the work inside the body. They are responsible for building, operating and regulating all tissues and organs. They often work together to accomplish their tasks in processes called protein-protein interactions.

In over 97 per cent of people with ALS, TDP-43 protein collects outside of its normal place in the nucleus of motor neurons and aggregates in the cytoplasm. There is still much to learn about this abnormal process and why it occurs in ALS.

With this grant, Dr. Mohan Babu is collaborating with Dr. Christine Vande Velde at the Université de Montréal and Dr. Antonia Ratti at the University of Milan, Italy. They will examine the protein-protein interactions that occur in different people with the same TDP-43 mutation to see if any unique patterns in the biology might explain each person’s specific clinical symptoms. They will grow motor neurons from stem cells derived from the skin and blood cells of people living with and without ALS and use powerful computer analysis techniques to analyze the complex interactions between TDP-43 protein and its surrounding proteins.

Learning whether TDP-43 interactions with other proteins can reflect unique aspects of ALS symptoms and the resulting clinical journey could be critical to advancing our understanding and treatment of disease in the years ahead.

How do environmental marks on RNA play a role in how ALS is caused?

Defining m6A epitranscriptomic variants in the context of two ALS genetic risk factors: TARDBP and C9ORF72

$100,000 awarded to Dr. Patrick Dion at McGill University

DNA holds the master code of genetic instructions for all processes that take place in the body. Another genetic molecule called RNA is created from those instructions and performs many important cellular processes including overseeing the production of proteins.

Environmental impacts can make changes to our DNA that affect how our genes are expressed. For example, environmental stress can add tags in specific locations on DNA in a process called methylation. This process modifies the genetic instructions, which in turn changes how much RNA and protein is produced.

Scientists have recently discovered that RNA can also be tagged by methylation, affecting how it functions in cellular health. This exciting new scientific field is called epitranscriptomics.

For this pilot project, Dr. Patrick Dion is collaborating with Dr. Guy Rouleau at McGill University. They will be among the first scientists to study RNA methylation in ALS. They will investigate a specific type of methylation called m6A in the RNA of cells that have mutations in TDP-43 and C9ORF72 genes, two of the most common genetic forms of ALS. m6A is the most abundant modification, and it affects almost all RNA processes.

Dr. Dion hopes to uncover new understandings about how environmental impacts affect RNA processes in ALS. This project promises to set the stage for future ALS research in an exciting new field.

Does prior exposure to common viruses influence ALS onset and disease progression?

Defining how viral infection influences the onset and progression of ALS

$100,000 awarded to Dr. Matthew Miller at McMaster University

Over our lifetime, a variety of common viruses, such as the cold or flu virus, can make us ill. We recover from these infections, but scientists do not know yet if those events change our experiences with other diseases that develop later in life.

Mutations in a protein called senataxin have been shown to cause some forms of juvenile-onset ALS. In previous work, Dr. Miller identified that senataxin is also critical to managing how a cell responds to viral infection. Since this discovery, he has been exploring a potential connection between the body’s response mechanisms to viruses and the onset and disease progression of ALS.

With this grant, Dr. Miller will expose ALS model mice to two common viruses — influenza and herpes simplex virus 1 — and then study whether those experiences accelerate ALS disease processes later in life. If they do, he will look for a mechanism to explain how that acceleration occurs, by investigating whether immune responses were triggered and if those responses were associated with abnormal behaviour by proteins involved in ALS.

New understandings about a potential link between exposure to common viruses and accelerated ALS disease onset and progression may lead to a better understanding of how ALS is triggered, who may be susceptible and new ways to potentially treat the disease.

Does a substance in gut or oral bacteria influence the disease course of ALS?

Host microbiota-derived products in amyotrophic lateral sclerosis

$100,000 awarded to Dr. Minh Dang Nguyen at the University of Calgary

In recent years, scientists have learned that changes in gut bacteria can influence health. For example, studies have linked reduced quantities of good bacteria with a variety of health conditions, including irritable bowel syndrome, diabetes and eczema.

Some types of gut bacteria secrete a substance called lipopolysaccharide (LPS) that is known to stimulate the innate immune response. The innate immune response takes place when the body identifies foreign invaders as a threat, increases inflammation and initiates other defensive processes in an attempt to remove them.

For this project, Dr. Minh Dang Nguyen and Dr. Gerald Pfeffer at the University of Calgary will use ALS mouse models to study forms of LPS from five different gut bacteria. They will investigate whether these specific forms of LPS cause inflammation and affect ALS disease onset and progression. If they do change the course of ALS in mice, the researchers will examine inflammatory processes and the composition of the mice’s gut bacteria to understand the underlying processes.

To further examine a connection between resident bacteria and the disease, Dr. Nguyen and Dr. Pfeffer will also test saliva from 50 people with ALS and 50 people without ALS to look for differences in their oral bacteria. In the future, changes in oral bacteria could serve as a useful biomarker to identify changes in ALS disease progression or responses to treatments in clinical trials.

Can a new ALS mouse model provide important information for understanding and treating ALS?

Establishing MATR3 S85C knock-in mice as a preclinical model of ALS

$100,000 awarded to Dr. Jeehye Park at the Hospital for Sick Children (SickKids) Research Institute, Toronto

ALS researchers often work with mice that have been genetically modified to model ALS in humans. Yet, it can be a challenge to create mouse models with the right levels of mutated proteins to accurately represent the protein abnormalities found in people with ALS. Without the right levels, it can be difficult to tell if research findings resulted from the disease or the presence of too much protein. Previous-generation mouse models that make high levels of the mutant MATR3 protein may not accurately represent disease mechanisms.

Dr. Jeehye Park has produced a new mouse model of the MATR3 mutation using a modern gene-editing technique called CRISPR. CRISPR involves editing the mouse’s MATR3 gene directly, similar to cutting and pasting a letter in a misspelled word to correct a typo. These mice are showing ALS-like symptoms, including motor abnormalities, inflammation and misplacement of MATR3 protein from the nuclei of motor neurons into the cytoplasm, with some aggregation occurring as well. The development of these symptoms is a promising sign that these mice may be good models for studying ALS.

Dr. Park received an ALS Canada-Brain Canada Career Transition Award in 2016. This 2019 grant leverages the previous funding to provide continuing support that will allow Dr. Park to finish validating the MATR3 mice models of ALS. She hopes to publish her work so that ALS researchers around the world may use these models in future research to understand the underlying biology and mechanisms that contribute to ALS, which could help identify new treatment targets.

Are the same faulty nerve-muscle connections in ALS mice also occurring in humans?

Characterization of neuromuscular junctions in ALS patients: a proof of concept study

$100,000 awarded to Dr. Richard Robitaille at the Université de Montréal

One of the earliest signs of dysfunction in animal models of ALS is the disruption of the place where neurons connect to muscle, called the neuromuscular junction (NMJ). Dr. Robitaille is one of the world’s leading scientists in studying the NMJ. In recent years, he has discovered several abnormalities at the NMJ in ALS mouse models that may help us better understand the disease while also identifying potential new avenues for treatment.

However, it is difficult to know if these findings in mice are the same in humans. With this grant, Dr. Robitaille aims to explore this for the first time. As part of a multi-disciplinary team that includes neurologist, Dr. Angela Genge, neurosurgeons and protein scientists, he will obtain small biopsies of muscle from people living with ALS and study the structure and function of the NMJs with the same methods used in the mouse models. Dr. Robitaille will also compare them to samples previously obtained from individuals without ALS. In addition, he will determine the levels of NMJ proteins in the biopsies to see if the changes mimic those observed in mice.

If this project confirms that human NMJs replicate the disease processes seen in ALS mice, the findings will significantly advance the understanding of human ALS and likely identify exciting new targets for treatments that could move quickly to clinical trials.

What is the role of the annexin A11 gene in ALS disease processes?

Investigating the mechanisms regulating the role of the ALS-associated annexin A11 protein in intraneuronal trafficking of RNP granules

$100,000 awarded to Dr. Peter St. George-Hyslop at the University of Toronto

Neurons are like long, living wires that connect to each other. They have an axon at one end and dendrites at the other. Electrical signals flow from the axon of one neuron to the dendrites of another neuron across gaps called synapses. For a healthy signal to cross the synapse, the right biological processes must occur in the axon and the dendrites. In ALS, motor neurons gradually break down and healthy signals no longer communicate with muscles.

In 2017, researchers discovered that mutations in a gene called annexin A11 (ANXA11) can be found in ALS. The protein expressed by this gene, called annexin A11, plays several roles in the body, including moving proteins to the right locations.

Recently, Dr. Peter St. George-Hyslop was a co-author of a study led by Dr. Michael Ward at the National Institute of Neurological Disorders and Stroke in the United States. They discovered that the annexin A11 protein acts like a seat belt to strap RNA and RNA-binding proteins to other cellular structures called lysosomes. This tethering allows the RNA and RNA-binding proteins to “hitchhike” to their final destinations in dendrites and axons, where they play an important role in maintaining their health. When annexin A11 protein is mutated, this tethering process malfunctions, impairing the normal movement of RNA and RNA binding proteins.

With this award, Dr. St. George-Hyslop will further explore how ANXA11 affects the proper biology of motor neurons. Findings from this project may provide new insights about the underlying mechanisms driving the development of ALS and potentially discover new treatment targets that help RNA and RNA binding proteins move to where they are needed.

Can advanced technology reveal the role of multiple cell types affecting ALS in humans?

Comparing the transcriptomes of motor neurons, astrocytes and microglia from spinal cords of ALS and non-ALS individuals

$98,400 awarded to Dr. Stefano Stifani at the Montreal Neurological Institute at McGill University

The primary symptoms of ALS are caused by degeneration of motor neurons. Abnormalities in other cells called glia that surround motor neurons also play an underlying role in ALS disease processes. Therefore, understanding the unique contribution of each cell type is important for developing effective treatments.

For this project, Dr. Stefano Stifani will collaborate with three colleagues at McGill University: Dr. Guy Rouleau, Dr. Carlo Santaguida and Dr. Luke Healy. Using a powerful new technology called single-cell RNA sequencing, they will focus on motor neurons, as well as two types of glia called astrocytes and microglia.

Dr. Stifani and colleagues will analyze spinal cord tissue generously donated by ten people with ALS and ten people without ALS. What sets this project apart from other ALS research using single-cell RNA sequencing is that these scientists will use spinal cord tissue that has not been previously frozen. The Montreal Neurological Institute has set up a program that allows scientists to analyze unfrozen autopsy tissue from people who choose to donate their bodies to scientific research.

The researchers believe that unfrozen spinal cord tissue may be better for research purposes because freezing and thawing tissue samples can cause changes that are not due to disease processes. New insights about disrupted biological processes in cells surrounding motor neurons may provide new information about ALS.

Can earlier palliative care consultation improve patient and caregiver quality of life?

Understanding palliative care and end-of-life needs in the ALS population: The first step to improving patient and caregiver quality of life

$55,437 awarded to Dr. Jocelyn Zwicker and Dr. Christine Watt at The Ottawa Hospital

Currently, there is no cure for ALS. It is a progressive disease that has an average life expectancy of two to five years after diagnosis.

Palliative care is an area of medicine aimed at improving the quality of life of patients and their families facing the problems associated with a life-threatening illness. These consultations can provide increased psychological support, assistance with advanced care planning and information on end-of-life symptoms and care. Typically, palliative care physicians are not involved as part of routine multidisciplinary care until later in the disease, or in some cases, not at all.

With this grant, Dr. Jocelyn Zwicker and Dr. Christine Watt will identify palliative needs and study the impact of offering earlier palliative care to individuals with ALS and their caregivers. The researchers will measure the benefits of palliative care by asking participants to complete surveys about their quality of life and mood at the time of the initial consultation, and then one month, three months and every three months after that. The surveys include the ALS-Specific Quality of Life Scale-Revised (ALSSQOL-R) and the Hospital Anxiety and Depression Scale (HADS). Individuals with ALS and their caregivers who decline early palliative care will have the opportunity to receive it at a later time.

The research team includes Dr. Usha Buenger, Dr. Ariel Breiner, Dr. Jill Rice, and Susan McNeely, RN at The Ottawa Hospital. In addition to assessing the feasibility and impact of providing earlier palliative care, the researchers also hope to identify the timeframe when people with ALS and their caregivers are most likely to benefit, and how to recognize individuals who may benefit from earlier consultation.

Can speech-recognition technology help diagnose ALS?

Machine learning in the detection of upper and lower motor neuron features in speech of patients with ALS

$100,000, in partnership with Orangetheory Fitness Canada, awarded to Dr. Yana Yunusova at the Sunnybrook Research Institute, Toronto

At present, no one test or procedure can diagnose ALS. Doctors focus on ruling out other diseases that share some similar initial symptoms. The complexity of the disease and the need for additional diagnostic tests means that for some people, there is a significant delay in making a definitive diagnosis.

Thirty per cent of people with ALS have bulbar-onset ALS. They experience voice and speech changes at disease onset due to a loss of motor neuron function in the corticobulbar area, the area of the brain that controls the muscles of the face, head and neck. Almost all other people living with ALS will have voice and speech difficulties at later stages of the disease.

For this project, Dr. Yana Yunusova will collaborate with Dr. Agessandro Abrahao and Dr. Lorne Zinman at Sunnybrook Health Sciences Centre in Toronto, Dr. Babak Taati at the UHN-Toronto Rehabilitation Institute, and Dr. Sanjay Kalra at the University of Alberta.

Dr. Yunusova and colleagues will use machine learning to train an artificial intelligence (AI) model on voice recordings from people with ALS. They will use voice recordings from people with primary lateral sclerosis, which affects mostly upper motor neurons, people with Kennedy’s disease, which involves only lower motor neurons, and people with ALS affecting both the upper and lower motor neurons. The goal is to create a tool that can separately identify the subtle acoustic features of upper and lower motor neuron disease.

The researchers hope that the new tool will assist existing methods in providing a faster and more accurate way to diagnose people with ALS. It may also help to determine the onset of bulbar symptoms in people with limb-onset disease. Further, it may provide a fast way to identify people who are carriers of specific gene mutations, allowing them to access clinical trials or proven therapies more quickly in the future.

ALS Canada Trainee Awards

Doctoral Award

Can an animal model provide new insights into the formation of stress granules?

Investigation of the interplay between stress and genetics in ALS

$75,000 awarded to Alicia Dubinski, a Ph.D. student in Dr. Christine Vande Velde’s lab at the Université de Montréal.

Dr. Vande Velde’s lab and her team have been investigating whether reduced levels of TDP-43 in the cell nucleus, as commonly seen in ALS, causes reduced levels of another protein called G3BP1. It is an essential protein for the formation of stress granules, protective structures that healthy cells make when they are exposed to environmental stress. Stress granules protect vulnerable RNA, molecules that translate genetic instructions and oversee protein production from becoming damaged. In ALS, disruption of proper stress granule biology appears to play a central role in the disease process. So far, the research on these structures has largely been conducted in cell studies.

With this grant, Alicia Dubinski will be among the first researchers to look at stress granule processes in an ALS animal model. She will apply mild heat to TDP-43 model ALS mice, similar to placing them in a warm sauna, and examine how the disease processes affect the formation of stress granules. She will also investigate how aging and being exposed to heat over time changes their ability to properly make stress granules.

This project builds on Dr. Vande Velde’s 2015 ALS Canada-Brain Canada Arthur J. Hudson Translational Team Grant. Continued support will allow Dubinski to shed light on how TDP-43 affects the formation of stress granules. New insights about this biological process may help identify new treatment targets for maintaining healthy formation of stress granules to halt or slow disease progression in ALS.

What is the role of a newly discovered protein in ALS?

Biological and pathological relevance of hnRNP A1B; an alternative splice variant of TDP-43

$75,000, in partnership with La Fondation Vincent Bourque and Brain Canada, awarded to Myriam Gagné, a Ph.D. student in Dr. Christine Vande Velde’s lab at the Université de Montréal.

In 97 per cent of ALS cases and nearly half of cases of frontotemporal dementia, the TDP-43 protein is misplaced to an area outside the cell nucleus of the motor neuron called the cytoplasm. The mislocated TDP-43 may be toxic to motor neurons because it aggregates in the cytoplasm, or because it is no longer performing its normal function in the nucleus.

Scientists have also discovered that mutations in another protein, called hnRNP A1, can cause ALS. In 2018, Dr. Christine Vande Velde discovered that when TDP-43 moves out of the cell nucleus, a new version of hnRNP A1 forms, called hnRNP A1B. Her preliminary work has shown that this new protein may be even more toxic and appears to have a greater ability to form potentially toxic clumps.

With this grant, Myriam Gagné will aim to understand how hnRNP A1B is involved in ALS disease processes by performing cell and mice experiments. She will also validate her findings using ALS tissue samples generously donated by people who had ALS.

Gagné hopes that new insights about the role of hbnRNP A1B in ALS may reveal a better understanding of disease processes and potential targets for new therapies and biomarkers.

Is the loss of normal function of C9ORF72 in a particular cell type a key driver of ALS disease processes?

Functional role and diagnostic signature of C9ORF72 in amyotrophic lateral sclerosis

$75,000 awarded to Rahul Kumar , a Ph.D. student in Dr. Peter McPherson ’s lab at the Montreal Neurological Institute at McGill University.

In 2011, scientists discovered that mutations in a gene called C9ORF72 are the most common genetic cause of ALS. People with these gene mutations make less of the normal C9ORF72 protein, but also have more toxic substances in their motor neurons.

Exactly how C9ORF72 mutations cause ALS remains unknown. Some researchers believe the main issue is the loss of normal protein function in motor neurons, while others believe toxic substances aggregating in the nucleus and cytoplasm cause problems.

Through an initiative called the ALS Reproducible Antibody Platform, co-funded by ALS Canada, the ALS Association (US) and the MND Association (UK), a 2016 ALS Canada Trainee Award recipient, Dr. Carl Laflamme discovered that methods used to detect C9ORF72 protein in some previous studies were not correct. Using new methods, he found that C9ORF72 protein was very concentrated in blood cells called macrophages, white blood cells that engulf and digest cellular debris.

For this project, Rahul Kumar will perform a set of experiments to find evidence for the loss of function theory by studying blood macrophages from people living with ALS. He will examine the levels of C9ORF72 protein using blood samples donated by families with familial ALS and investigate how C9ORF72 in macrophages affects the normal movement of substances inside cells, a proposed normal function for this protein.

If the loss of normal function of C9ORF72 protein is essential in ALS disease processes, that insight will provide important information for developing new ALS treatments in the future.

Is an experimental drug that can prevent abnormal protein behaviour in ALS already out there?

Validation of small molecules preventing TDP-43 aggregation as a therapeutic for amyotrophic lateral sclerosis

$75,000, in partnership with La Fondation Vincent Bourque and Brain Canada, awarded to Marc Shenouda, a Ph.D. student in Dr. Janice Robertson’s lab at the University of Toronto.

TDP-43 is a protein that behaves abnormally in the motor neurons of 97 per cent of people with ALS. It is usually found in the nucleus, but in people with ALS, it becomes trapped outside in the cytoplasm where it forms aggregates. One theory is that these aggregates are toxic to motor neurons. Therefore, preventing these clumps from forming, or breaking them apart, could be a potential method for treating ALS.

In Dr. Robertson’s lab, Marc Shenouda has been using a computer program that models how compounds fit together with proteins. He has screened 50,000 experimental drugs to look for those that have the most potential to bind to TDP-43 and prevent it from aggregating. He has created a shortlist of 500 drug candidates for further testing.

For this project, Shenouda will test 500 experimental drugs in motor neuron cells in culture to see if they can effectively reduce TDP-43 aggregation and toxicity. He will then take the best candidate compound and test it in a TDP-43 ALS mouse model to see if it can change the course of the disease. Shenouda hopes that this project may discover a potential new treatment for ALS that could someday be tested further in a clinical trial.

Could newly discovered tags on TDP-43 protein explain its abnormal behaviour in ALS?

Disrupted SUMOylation facilitates rogue TDP-43 in ALS

$75,000, in partnership with Brain Canada, awarded to Terry Suk, a Ph.D. student in Dr. Maxime Rousseaux’s lab at the University of Ottawa.

Proteins are large molecules that play many essential roles in the body. They do most of the work inside cells and are necessary for the structure, function and regulation of all tissues and organs. TDP-43 is a protein that is usually found inside the cell nucleus. However, in most people with ALS, it is located outside the cell nucleus in the cytoplasm of motor neurons.

Tags called small ubiquitin-like modifiers (SUMOs) can change how proteins function and where they are located in a cell. Scientists have also found SUMOs on TDP-43 protein, but nobody knows yet how these tags might change TDP-43 structure, location, or function.

With this grant, Terry Suk will investigate exactly where SUMOs tag TDP-43 and how they affect its behaviour using fruit fly models. He will also examine donated post-mortem spinal cord tissue from people who had ALS to learn about the extent of SUMO tagging of TDP-43 in ALS disease processes.

New learning about how SUMO tagging affects TDP-43 and ALS disease processes may lead to a new therapeutic target for ALS in the future.

Postdoctoral Fellowship

Can new understandings about nuclear speckles lead to new treatment options for ALS?

Regulation of TDP-43 and FUS by phosphoinositides in nuclear speckles during ALS

$165,000 awarded to Dr. Ulises Rodríguez Corona, a postdoctoral fellow in Dr. Marlene Oeffinger’s lab at the Institut de recherches cliniques de Montréal (ICRM).

Inside a cell nucleus, many biological processes take place. Two proteins, TDP-43 and FUS, are primarily found inside the nucleus where they play important roles. In ALS, they are mutated and behave abnormally by moving out of the nucleus and aggregating in the cytoplasm. The mislocation of TDP-43 outside the cell nucleus is a hallmark sign of most ALS cases.

Scientists have recently discovered other structures inside the cell nucleus. But instead of having distinct shapes and cell walls, they resemble droplets of liquid that form and dissolve. These liquid droplets, called nuclear speckles, contain proteins and other molecules called RNA that can change protein behaviour.

Preliminary work by Dr. Ulises Rodríguez Corona has shown that specialized fats, called nuclear phosphoinositides (PIs), may stick to TDP-43 and FUS inside nuclear speckles, changing how these proteins behave and move.

With this grant, Dr. Rodríguez Corona will investigate whether PIs stick to TDP-43 and FUS proteins and how that changes their function and movement into and out of nuclear speckles. New understandings about the normal processes that direct TDP-43 and FUS proteins to move and function may lead to potential new treatment options for ALS by normalizing their behaviour.