Projects Funded 2016
Table of Contents
ALS Canada-Brain Canada Arthur J. Hudson Translational Team Grant
The projects that have received Hudson Grants in 2016 are:
Pathogenic mechanism of C9orf72 haploinsufficiency in ALS/FTLD: a road to therapeutic discovery
Principal Investigator: Dr. Janice Robertson, Professor, University of Toronto
$1.6 million over five years
- Dr. Julia Keith, Neuropathologist, Sunnybrook Health Sciences Centre
- Dr. Ian Mackenzie, Professor, University of British Columbia
- Dr. Peter McPherson, Professor, McGill University
- Dr. Ekaterina Rogaeva, Professor, University of Toronto
- Dr. Lorne Zinman, Neurologist, Sunnybrook Health Sciences Centre
In 2011, scientists discovered that hereditary mutation of a previously unstudied gene called C9ORF72 was the most prominent cause of both ALS and frontotemporal dementia (FTD). These mutations were found in both hereditary (familial) and non-hereditary (sporadic) forms of the disease and comprise about one third of all cases.
Since that discovery, ALS researchers have aimed to understand the normal function of C9ORF72, determine how mutations in the gene can cause the disease and create animal and cellular models to study it. In 2015, Dr. Janice Robertson published a landmark paper that demonstrated the existence of two different forms of C9ORF72 protein, including one that localized to the outer layer of the cell’s command centre called the nucleus. This was the first indication of what would become a breakthrough discovery by three teams independently showing that mutations in C9ORF72 disrupt the movements of critical substances into and out of the nucleus.
When a mutation in a gene causes ALS, it may do so by removing the normal function of the resulting protein (loss of function), by causing the resulting protein to have some extra, toxic function (gain of function), or in some cases, both. The recent discovery of the effects on movement in and out of the nucleus has led to a major focus on the gain of function hypothesis. However, some work has indicated that loss of function of C9ORF72 may also be important in ALS and this has still not been well defined. Furthermore, many studies have indicated that there is a reduced amount of C9ORF72 in affected regions, supporting the loss of function hypothesis.
In this Hudson project, the team led by Dr. Robertson will pursue a comprehensive understanding of the normal functions of C9ORF72 and provide a better determination of whether loss of these functions via mutation may cause or contribute to ALS.
One of the first discoveries about C9ORF72 was that it was a type of protein containing something called a DENN domain, which means it has function in normal movement of substances inside cells. Team member Dr. Peter McPherson, at McGill University, who discovered this unique function for DENN proteins, will further explore preliminary data that connects C9ORF72 to critical compartments in cells called lysosomes and their ability to recycle cellular components through a disposal process called autophagy. In recent years, numerous ALS genes have been connected to autophagy pathways.
Furthermore, the team will examine other preliminary work showing that normal C9ORF72 may be necessary for proper movement of substances into and out of the nucleus, indicating that mutations in this gene causing ALS might have a dual effect on cells through loss and gain of function mechanisms. Finally, the team will study a completely unexplored potential function of C9ORF72 in a process called compensatory collateral sprouting, which is a function of neurons where they can regrow new connections at their ends when they get disconnected from muscles. Other work, including that of Canadian researchers, has indicated that this sprouting is impaired in ALS, but connection to C9ORF72 has not yet been made.
Nothing has been a greater focus of global ALS research over the past few years than C9ORF72 and the majority of the work has emphasized studies centred on gain of function hypotheses. When a mutation causes an extra, toxic function, the first therapeutic strategy is to use advancements in medicine that are capable of reducing the amount of the mutant protein. However, without first understanding whether the normal function of C9ORF72 is crucial to the health of motor neurons and other cell types affected in ALS, such strategies may not be effective, or if mutations cause loss of function, potentially detrimental. This Canadian team is uniquely qualified to comprehensively answer these questions and their findings will ultimately enhance our ability to efficiently target C9ORF72 and its mechanisms in future treatment strategies.
A patient-derived IPSC platform of disease relevant cell models for biological studies
Principal Investigator: Dr. Guy Rouleau, Professor, McGill University
$2.2 million over five years
- Dr. Benoit Coulombe, Université de Montréal
- Dr. Patrick Dion, McGill University
- Dr. Edward Fon, McGill University
- Dr. Jack Puymirat, Université Laval
- Dr. Stefano Stifani, McGill University
One of the greatest advancements in science over the past 10 years is the development of induced pluripotent stem cells (iPSCs). The inventors were awarded the Nobel Prize in Medicine in 2012. These cells make it possible to create essentially any cell type in the body using any other as a starting point. In ALS, iPSCs provide the opportunity to take skin or blood cells from someone living with the disease and turn them into motor neurons and other relevant cell types so that we may study human cells with the exact genetic makeup of the person donating them.
In this Hudson project, the team will create iPSCs from 40 people living with ALS, including individuals with different hereditary genetic mutations, those with typical disease progression and those with atypical disease features. These iPSCs will them be made into motor neurons and astrocytes (the personal assistant cells to motor neurons that are also actively malfunctioning in ALS). Furthermore, using a special technology called CRISPR-Cas9, the team will specifically change the mutations in those from hereditary cases back to the correct form so that they may also be studied alongside those with the mutations. These are called isogenic controls and are a highly sought after resource in the global ALS research community.
Using these newly developed motor neurons and astrocytes, the team will first determine if they develop any specific characteristics that can be correlated with the disease course and examine if they show differences as compared to others in the study. Furthermore, they will use these cells to develop novel tests for screening drug treatments that may be able to indicate an ability to slow down disease processes in human motor neurons.
Finally, the team will also take advantage of this unique model of ALS to learn more about how the disease occurs. Using a technique called quantitative proteomics, they will examine all of the substances involved in the communication between human motor neurons and astrocytes in these lab cultures. It is known in ALS that this crosstalk can be critical to the development and possibly progression of the disease and gaining a full understanding of how this happens will undoubtedly reveal new targets to treat the disease.
The outcomes of this intense five-year collaboration will not only provide a better understanding of ALS and potentially new options to pursue for treatment, but will also develop a valuable new set of tools to be shared amongst the global ALS community to impact the international effort in making it a treatable, not terminal disease.
ALS Canada-Brain Canada Discovery Grants
The 2016 recipients of ALS Canada- Brain Canada Discovery Grants are:
Gary Armstrong, PhD
Montreal Neurological Institute, McGill University
Title: Generating expanded repeats in the C9ORF72 ortholog in zebrafish
Over the past several years, zebrafish have proven to be a valuable model to study motor neuron degeneration and search for new treatments. To date, several key mutations that cause ALS in humans (SOD1, TDP-43, FUS, etc.) have been introduced to zebrafish resulting in motor neuron dysfunction and paralysis. These models not only provide an opportunity to study disease mechanisms with a goal of better understanding ALS, but they are also important tools for screening potential therapeutics to see if motor dysfunction can be slowed down using systems designed to robotically test thousands of substances in a single experiment. A substance is applied to the water they swim in and they are examined to see if impairment is slowed.
In 2011, the most common ALS mutation was discovered as an abnormally long piece of DNA in a gene called C9ORF72. To date, some difficulty has arisen in the creation of C9ORF72 zebrafish models that resemble the actual genetic abnormality in humans. Through this Discovery Grant, Dr. Gary Armstrong will use an advanced gene editing technique called CRISPR/Cas9, including a novel modification he invented, to more accurately accomplish this goal. If successful, Dr. Armstrong’s zebrafish could represent a very valuable resource for the ALS research community and provide an important new tool for therapeutic screening.
Dr. Neil Cashman
University of British Columbia
Title: Mechanisms of in vivo synaptic transmission of mis-folded human SOD1
The process by which ALS spreads throughout the body remains a mystery, but Dr. Neil Cashman has spent decades looking at how abnormal changes in shape (scientifically called misfolding) of crucial proteins (the substances that do life processes) in our cells might propagate disease from one cell to another. In particular for ALS, a protein called superoxide dismutase 1 (SOD1) has been hypothesized as the culprit that misfolds and propagates disease by triggering a domino effect of further SOD1 misfolding.
Signals from our brain to our muscles and throughout our nervous system occur through internal wiring of cells called neurons that are interconnected. Between neurons, there is a tiny gap at their connection point called the synapse and to date, there has been no demonstration of misfolded SOD1 being able to cross the synapse to spread toxicity from cell to cell. Using unique aspects of fruit fly (Drosophila) neurons involved in smell, Dr. Cashman has teamed up with a Drosophila expert, Dr. Catherine Cowan, also at UBC, to visualize if this neuronal transmission indeed occurs. If so, Dr. Cashman intends to further examine the characteristics of how SOD1 can cross synapses and even determine if he can develop a test in flies that would allow for screening of drugs that may be able to alter this transmission and possibly be a blocker of ALS spread throughout the body. Ultimately, if the hypothesis is true, the implications would be huge for ALS treatment. Proof that SOD1 misfolding is common to most cases of ALS, and not just the 2% that have hereditary SOD1 mutations, will massively increase the potential value of SOD1 targeted therapeutics that are already in the clinical trial pipeline.
Dr. Charles Krieger
Simon Fraser University
Title: Impaired neuromuscular junction connectivity in amyotrophic lateral sclerosis
Many scientists believe that one of the earliest abnormalities in ALS is dysfunction at the place where motor neurons connect to muscle, known as the neuromuscular junction (NMJ). Dr. Charles Krieger has observed that a substance called adducin, which is critical to the structure of the NMJ, is altered in multiple animal models of ALS. As a result, Dr. Krieger has hypothesized that abnormal levels or regulation of adducin may be responsible for this early NMJ dysfunction.
In this Discovery Grant, Dr. Krieger will collaborate with NMJ expert, Dr. Richard Robitaille of the Université de Montréal to further examine the role of adducin in ALS. Using two different animal models (fruit flies and mice), representing two different genetic causes of ALS (TDP-43 and SOD1), they will examine the mechanisms of adducin alteration by looking at its regulation and the various proteins it interacts with to properly maintain NMJ structure. Boosting NMJ health may represent an important avenue to slow down ALS and unraveling the role of adducin in maintaining this may provide unexplored new targets for treatment.
Eric Lécuyer, PhD
Université de Montréal
Title: Defining conserved functions of RNA binding proteins in stress-granule biogenesis
RNA is a mobile form of genetic information that is made from our DNA and its complex regulation is extremely important to the proper functioning of our cells. Since the discovery of TDP-43 in 2006 as a substance that plays a major role in ALS, the hypothesis that abnormal regulation of RNA is critical to the disease has come to the forefront. In the past decade, this has only strengthened as numerous newly-identified ALS genes pointed to an influence on RNA biology.
One of the standard features of RNA biology is the formation and disintegration of structures called stress granules that are created temporarily to protect RNA and RNA-binding proteins in times of potentially harmful environmental triggers. However, in ALS it has been observed that stress granules fail to properly break apart. In this Discovery Grant, Dr. Eric Lécuyer will use an unprecedented library of tools that recognize RNA binding proteins in stress granules to better understand their content in human stem cells derived from people living with ALS. Furthermore, Dr. Lécuyer will use a second set of tools developed in his lab to systematically remove RNA binding proteins from stress granules to determine which ones are critical to the proper formation and disintegration, and finally, determine how they play a role in toxicity to motor neurons in ALS.
An earlier Discovery Grant and Hudson Grant team have focused on understanding the content and dynamics of stress granules in ALS using other methods and the work of Dr. Lécuyer should complement these efforts well in a way that brings strong expertise into an already world-leading team effort in Canada. The magnitude of evidence that abnormal RNA biology is a key factor in how ALS is caused has grown considerably in recent years and finding ways to normalize areas of malfunction may represent one of the best avenues for discovering treatments that are significantly effective in slowing down the disease.
Marlene Oeffinger, PhD
Institut de recherches cliniques de Montréal (IRCM)
Title: Proteomic and transcriptomic profiling of paraspeckle function in healthy and ALS model neuronal cells
Over the past decade, the number of new cellular structures that have been identified with critical functions for life has expanded considerably. One of the more recent discoveries is a structure called a paraspeckle, which remains poorly understood but also has multiple components that have been identified as proteins that can cause ALS when altered through mutation, and may disrupt critical RNA biology that has been strongly linked to the mechanism of disease.
Using advanced protein and RNA identification techniques, Dr. Marlene Oeffinger will examine the full contents of paraspeckles to characterize them in a way that has not previously been done in the nervous system. Furthermore, her lab will observe how paraspeckles function and are altered in neuronal cells that have ALS-causing mutations. By tackling this as-yet unstudied cellular structure in ALS, Dr. Oeffinger may not only better understand how the disease occurs, but also potentially identify new and exciting treatment targets.
Alex Parker, PhD
CRCHUM, Université de Montréal
Title: Investigation of microbiota mediated suppression of motor neuron degeneration in genetic models of ALS
Scientists have long wondered what the contribution of environment is to ALS and in recent years the idea that a combination of genetic susceptibility and environmental triggers has taken shape. Furthermore, one of the emerging areas of interest for environmental exposure is the gastrointestinal tract and how dietary exposures might influence our nervous system through something called the gut-brain axis.
Dr. Alex Parker is an expert on studying the biology of tiny worms called C. elegans, including using them as animal models to understand ALS and screen for potential treatments. When worms are altered to have a mutant gene that causes ALS in humans, they get motor neuron degeneration and paralysis. Through a partnership with a company aimed at studying fat accumulation when worms were exposed to probiotics (helpful bacteria, as in certain yogurts, that are considered healthy for your digestive system) in their food, Dr. Parker serendipitously discovered that feeding the worms some of these probiotics also slowed down progression of symptoms in ALS worms. In this Discovery Grant, Dr. Parker will more closely examine how this neuroprotection occurs and what the underlying biology is that explains the effect. Understanding how dietary probiotics might alter cellular mechanisms in worm that may be protective in ALS may not only tell us more about the disease and the influence of environmental factors, but may also reveal information about the potential for interesting new types of experimental treatments.
Lisa Topolnik, PhD
Centre Hospitalier de l’Université Laval (CHUL)
Title: Decoding motor cortex circuit abnormalities at ALS onset through combined two-photon imaging in vivo and pharmacogenetics
ALS is a disease that is characterized by degeneration of both upper and lower motor neurons, and while a significant proportion of work has focused on motor neurons in the spinal cord, the motor cortex in the brain, where electrical signals to your muscles originate have clear, but poorly understood involvement in the disease process. One of the hypotheses in ALS is that excessive activity of upper motor neurons may be an early sign of the disease and Dr. Lisa Topolnik, an expert in brain circuitry (understanding how the various brain neurons interact), has developed a unique system to study whether this is a result of abnormal actions of other neurons (called interneurons) in the motor cortex that should be moderating this neuron firing. In this Discovery Grant, Dr. Topolnik will use brain imaging to examine the effects in motor cortex when ALS model mice are using their muscles prior to and at onset of disease symptoms. If the hypothesis that these interneurons are failing to inhibit excessive upper motor neuron activity is true, Dr. Topolnik will examine the effect of various treatments to circumvent this effect. Ultimately this is an example of how expertise outside the field of ALS can be made applicable to answer key questions that have not been previously examined and this work will undoubtedly yield new insight into how the disease is caused, with further potential to discover novel treatments as well.
ALS Canada-Brain Canada Career Transition Award
The recipients of the 2016 ALS Canada-Brain Canada Career Transition Award are:
Dr. Jeehye Park
Assistant Professor, Department of Molecular Genetics
Hospital for Sick Children, Toronto, ON
Title: Characterization of MATR3 mutations associated with ALS
$315,000 over three years
Dr. Park has made significant contributions to neurodegenerative disease research since the beginning of her career. During her PhD work in South Korea with Dr. Jongkyeong Chung, Dr. Park discovered a key connection between two Parkinson’s disease pathways that had a major impact on the field and was published in the elite scientific journal Nature. She subsequently pursued postdoctoral research at Baylor College of Medicine under the guidance of Dr. Huda Zoghbi, where Dr. Park helped to create a network of laboratories with expertise across different animal models to screen for treatments for the neurodegenerative disease spinocerebellar ataxia 1, which led to yet another paper in Nature. Her research then led her to study RNA binding proteins (RBP), where she not only developed a new tool to study them, but became interested in the multiple RBPs that are linked to ALS.
In her lab, Dr. Park will examine how abnormalities in RBPs – in particular, one called Matrin 3 (MATR3) – can lead to ALS. MATR3 was discovered to be a genetic cause of ALS in 2014 and has yet to be studied in any detail. By creating the first-ever cell, fruit fly and mouse models of MATR3, Dr. Park will learn both about the functions of MATR3 and how mutations can confer motor neuron degeneration. Dr. Park will then search for other genes that may increase or reduce mutant MATR3 toxicity in both human cells and fruit fly models to find potential targets for treatment, and follow up with the most promising candidates being tested in the new MATR3 mouse models with an aim to eventually move them forward translationally into the clinic.
As a member of the Canadian ALS research community, Dr. Park will be able to integrate the knowledge gained about MATR3 with the work of others here and around the world as yet another puzzle piece in understanding ALS. By focusing the early stages of her independent career on a less understood ALS mechanism, she intends to find connections between MATR3 and more prominently studied RBPs like TDP-43 and FUS to ultimately unravel key mechanisms in the development of ALS, as well as new targets to treat the disease.
Dr. Veronique Belzil
Mayo Clinic, Jacksonville, Florida
Supervisor: Dr. Leonard Petrucelli
Title: Discovery of transcriptomic biomarkers and epigenetic therapeutic targets for c9ALS and sALS
$110,000 over two years; eligible for an additional $315,000 over three years
Dr. Belzil began her research career as a PhD student at the Université de Montréal under the guidance of world renowned geneticist and Director of the Montreal Neurological Institute and Hospital, Dr. Guy Rouleau. During this time, Dr. Belzil pursued a better understanding of the genetics behind familial/hereditary ALS and led or contributed to more than 20 manuscripts, an amazing accomplishment for a graduate student.
For the past four years, Dr. Belzil has spent her postdoctoral studies pursuing the complex understanding of how alterations in genetic regulation may lead to ALS not just in certain familial forms, but in sporadic ALS that makes up 90-95% of cases. She has led or contributed to a large number of important discoveries.
The high impact work that Dr. Belzil has been pursuing during her postdoctoral training translates very well into an expanded program for an independent laboratory and she aims to continue to tackle these mechanisms as an Assistant Professor. The program she has outlined is also designed to apply the knowledge of these discoveries into a strategy to develop novel and exciting new treatments for ALS that would be based on an intricate understanding of the disease.
Dr. Petrucelli and a mentoring committee at Mayo Clinic are committed to assisting Dr. Belzil to not only reach her goal of becoming an independent investigator at a Canadian institution, but to become an internationally recognized leader in translational ALS research.
Dr. Kessen Patten
Assistant Professor, Genetics and Neurodegenerative Disease
Centre INRS–Institut Armand-Frappier, Laval, QC
Title: Pathogenic mechanisms of C9ORF72 repeat expansion in ALS and development of therapeutics
$315,000 over three years
Dr. Patten started his research career as a PhD student at the University of Alberta under the supervision of Dr. Declan Ali in 2004. There he trained in electrophysiology, cell biology and imaging using zebrafish as a model to study neurodevelopment. After publishing several manuscripts on his discoveries and receiving multiple awards, including national recognition for the outstanding quality of his PhD thesis, Dr. Patten pursued a postdoctoral fellowship in Montreal with Drs. Florina Moldovan and Pierre Drapeau. During that time, among other achievements, he developed zebrafish models of human disease including ALS, and used those models to develop a high-throughput method for drug discovery. This procedure was then used by Dr. Patten in the identification of pimozide as a lead compound in a translational pipeline that has led to a multi-centre Canadian clinical trial to start in 2017. The trial is being supported by the first ALS Canada-Brain Canada Arthur J. Hudson Translational Team Grant that was awarded in 2014.
In the initial years of his independence as an Assistant Professor, Dr. Patten will pursue the development and use of zebrafish models of the most common genetic cause of ALS, C9ORF72, as well as use of the high-throughput screening method to examine more promising compounds for further examination. As a key addition to his work, he has formed strong collaborations with international ALS experts with proficiency in developing motor neurons from induced pluripotent stem cells (iPSCs) that will undoubtedly strengthen the ability to translate zebrafish discoveries to the clinic via the use of human cells.
Dr. Patten has been a regular attendee at the ALS Canada Research Forum for the past several years and has formed relationships with a number of other investigators in the community. Combined with multiple other Canadian investigators using ALS model zebrafish, C. elegans worms, Drosophila fruit flies, mice, rats and iPSC derived motor neurons, Dr. Patten will strengthen this country’s expertise on forming a pipeline of drug discovery that can efficiently reach the clinic and ultimately help make ALS a treatable, not terminal disease.
Ronald Peter Griggs and Tim E Noël Postdoctoral Fellowships
In 2016, we awarded two fellowships at $165,000 over three years:
Carl Laflamme, PhD
Montreal Neurological Institute, McGill University
Supervisor: Dr. Peter McPherson
Title: Characterization of the cell biological function of the ALS gene C9ORF72
In 2011, the most common genetic cause of ALS (and frontotemporal dementia) was discovered, identified as an abnormality in a gene called C9orf72. Prior to that, researchers did not know of C9orf72 and a great deal of effort has been put forth to understand what its function is so that we might understand how a mutation could cause the disease. One of the first aspects discovered about C9orf72 was that it contains something called a DENN (differentially expressed in normal and neoplastic cells) domain, which means it is capable of altering function of substances called Rabs. Rabs are key factors in the shuttling of important pieces to our cellular health between the surface of cells and the site of their action, and improper Rab functioning could cause many important processes to be disrupted. DENN domains were discovered in the laboratory of Dr. Peter McPherson at the Montreal Neurological Institute, McGill University.
ALS Canada is pleased to announce that the 2016 Ronald Peter Griggs Memorial Postdoctoral Fellowship in ALS Research was awarded to Dr. Carl Laflamme, who will study this function of C9orf72 as a potential mediator for membrane trafficking. Dr. Laflamme’s preliminary evidence has already demonstrated that C9orf72 controls the function of a specific Rab (Rab9), which already provided some crucial information as to C9orf72’s importance in the normal functioning of motor neurons and other cell types affected in ALS. As Dr. Laflamme further unravels these mechanisms, it is likely that new therapeutic targets will emerge that could be developed to make ALS a treatable disease.
Sali Farhan, PhD
Broad Institute of MIT and Harvard
Supervisor: Dr. Benjamin Neale and Dr. Mark Daly
Title: Leveraging large-scale exome sequencing data to elucidate the genetic influences of amyotrophic lateral sclerosis
Over the past two decades, the best way to study the molecular aspects of ALS has been through discovery of genetic causes of the familial/hereditary disease (which only accounts for 5-10% of cases) that can then be used in the laboratory to make cell and animal models that mimic the motor neuron degeneration and abnormal biology of the human disease. In the past five to ten years, there has been an explosion in the number of genetic causes discovered, each representing a new tool to utilize in understanding the disease and identifying new targets for treatment. However, some genetic causes still remain to be identified and they may be key pieces to a puzzle by linking to the others that are now known and becoming understood.
ALS Canada is pleased to announce that Canadian scientist Dr. Sali Farhan is the recipient of the 2016 Tim E. Noël Postdoctoral Fellowship to work on furthering our genetic understanding of ALS under the supervision of world class geneticists Dr. Benjamin Neale and Dr. Mark Daly at the Broad Institute of MIT and Harvard. During her work, Dr. Farhan will analyze one of the largest databases of familial ALS genetic sequences to potentially discover new genetic mutations that cause the disease, but also to confirm those that have been identified in other studies. Furthermore, the state-of-the-art techniques for these analyses will provide Dr. Farhan with valuable expertise that will be applicable in future endeavours that aim to understand the genetic underpinnings of sporadic ALS through whole genome sequencing. Through this study and those of the future, the training from this fellowship will poise Dr. Farhan to make significant advances to our understanding of ALS and stimulate new directions for identification of therapeutics.
ALS Canada Doctoral Research Awards
2016 recipients of ALS Canada doctoral research awards are:
Supervisor: François Gros-Louis
Title: Development and characterization of a new tissue-engineered skin model for the study of ALS
Most of the attention around making ALS treatable is placed on development of therapies that can slow down disease progression. Equally important is the ability to diagnose people earlier so that such therapies can be applied at a time when motor neuron loss is not already so significant. Researchers have long sought after something called biomarkers, which are substances that can be detected through imaging or in body fluids (blood, cerebrospinal fluid, etc.) that can indicate if someone has ALS at a time point earlier than diagnosis by exclusion. Ideally, these tests will be available at symptom onset, and possibly someday, before physical manifestations of ALS ever occur.
For decades, ALS clinicians and researchers have known that the skin of people with the disease has unique characteristics, but until recently no one had created a way to study these effectively. In recent years, Dr. François Gros-Louis utilized expertise of collaborators at Université Laval to a develop tissue-engineered skin model derived from people living with ALS. PhD student Bastien Paré has received a 2016 Doctoral Research Award from ALS Canada to study these models for biomarkers that may be detectable at or before the onset of neurological symptoms. In preliminary work, Bastien has already discovered intriguing abnormalities in a region called the extracellular matrix, and perhaps even more promising, clumps of TDP-43, a well-known marker of ALS, in the skin. Furthermore, Bastien will examine the possibility of tissue-engineered skin to be a model for monitoring the effectiveness of drug treatments, giving the model even more value. Ultimately, the potential to use a simple skin sample for diagnosis and/or monitoring of clinical trials is a very attractive notion and would be a very important part of making ALS a treatable. It will certainly be exciting to observe the progress of Bastien’s work in the years ahead.
The Hospital for Sick Children
Supervisor: Christopher Pearson
Title: Characterization of the disease-associated expanded C9orf72 repeat and potential therapeutic target
Genetic mutations can take on many forms. Some are deleted portions of a gene, some are simply wrong pieces substituting for the right ones and some are additions, where a gene becomes longer with excess, unnecessary pieces inserted, making it oversized and dysfunctional. In 2011, the most common genetic cause of ALS was discovered. This gene, called C9orf72, can be abnormally elongated, causing it to be dysfunctional and lead to motor neuron degeneration, but the exact mechanisms that confer ALS are not yet fully understood. When this gene grows in length, it forms unusual shapes that our natural defense mechanisms attempt to repair. However, it is believed that in the case of C9orf72, our DNA repair may actually make things worse.
With funding from ALS Canada, Monika Schmidt will pursue this topic for her PhD work in the lab of Dr. Christopher Pearson at The Hospital for Sick Children. Previous work in the Pearson lab has demonstrated that a substance in our DNA repair system, called MutSb, which is particularly good at fixing some types of genetic errors, is particularly bad at causing elongated gene mutations to have even worse effects, including making them even longer and possibly triggering disease.
Monika will examine how MutSb may play a role in making C9orf72 toxic, or in enhancing its toxicity by looking at their specific interaction. The work will include examining the connection both in laboratory cells and in a recently created mouse model of C9orf72 ALS, and could potentially identify MutSb as a therapeutic target for future treatment. Ultimately, Monika’s work represents a very novel avenue of research in the field and we look forward to learning of her results as they emerge.
ALS Canada Bridge Grants
2016 recipients of ALS Canada Bridge Grants are:
Dr. Neil Cashman, University of British Columbia
Dr. Cashman will receive additional support to work on the connections between two prominent ALS proteins, SOD1 and TDP-43. In ALS, TDP-43 is a protein that is abnormally shaped (called “misfolded”) and found in the wrong location in cells in 98% of cases. His hypothesis is that in sporadic ALS, this mislocalized, misfolded TDP-43 causes further misfolding of SOD1, and thus starts a domino effect of misfolded SOD1 and spreading of pathology throughout the body. Last year, he received a two year Bridge Grant to explore the following:
Though SOD1 and TDP-43 were the first two prominent genetic factors discovered in ALS, our understanding of their unique functions did not yield many studies examining a relationship between them in the ALS disease process. Dr. Cashman aims to first examine if abnormal structure of normal, non-mutant SOD1 (wild-type SOD1), which is termed “misfolding”, can cause neuronal death, as well as the mechanism by which this may occur. When SOD1 has a mutation, it misfolds and can cause familial (hereditary) ALS. Demonstration that misfolded wild-type forms can cause neurodegeneration would implicate SOD1 in sporadic ALS as well. Uniquely, he will then examine if mutations in TDP-43 can trigger wild-type SOD1 misfolding in cell cultures and possibly model ALS disease in a novel mouse model designed to study this connection.
To complement that work, Dr. Cashman will explore how a specific amino acid on TDP-43 called tryptophan might confer the ability to cause SOD1 misfolding. Proteins consist of long chains of substances called amino acids and Dr. Cashman had previously shown that a specific tryptophan amino acid is critical to SOD1 misfolding that he hypothesizes as starting a misfolding cascade that leads to ALS spread throughout the body. Recently he has determined that tryptophans on TDP-43 may also be key component to their ability to cause subsequent SOD1 misfolding. Using the 2016 Bridge funds, he will work to determine which tryptophan amino acids in the TDP-43 sequence are important for this function. If identified and his hypothesis is proven correct, this may represent a very plausible target for therapy, especially given modern technology of genetic modification (like CRISPR) that could alter these tryptophans to something innocuous.
Dr. Honglin Luo, University of British Columbia
Dr. Luo’s work is aimed at understanding if there is a role for enterovirus (EV) infection in the development of ALS. In work published last year, she found an intriguing set of similarities between motor neurons in ALS and cells infected with EV, including abnormal redistribution, clumping and cleavage of TDP-43. This disruption in TDP-43 is seen in 98% of ALS cases. As a result, Dr. Luo will test if chronic, persistent EV infection many contribute to or cause sporadic ALS by looking for EV presence in human ALS post-mortem tissue, determining if EV infection can accelerate or worsen disease in established ALS mouse models, and examining if mice making EV proteins develop ALS-like symptoms. If successful, Dr. Luo will then seek to identify the mechanisms of action as a means of developing therapeutic strategies.
Christopher Pearson, PhD, The Hospital for Sick Children
Dr. Christopher Pearson will receive another one year of support to continue work on the biology of the most prominent genetic cause of ALS, C9ORF72, which is mutated by having pieces of DNA abnormally replicated into long stretches that are dysfunctional. Through his work supported by a 2015 Bridge Grant, Dr. Pearson will now be able to extend his focus to learning more about why these stretches happen and focusing on a discovery of a particular substance called MutSβ that typically is very good at fixing damaged DNA, but can actually worsen situations where genes are abnormally elongated, as with C9ORF72. By working at both levels, Dr. Pearson’s lab should both gain important insight into how C9ORF72 mutations cause toxicity in ALS and also further develop MutSβ as a potential target for therapy. Dr. Pearson has spent his 16 year career focused on diseases caused by these specific elongation mutations and when the field first identified C9ORF72 as an ALS gene in 2011, he brought his expertise to the table in an effort to contribute knowledge that wasn’t pre-existing in the community.
Chantelle Sephton, PhD, Université Laval
Dr. Chantelle Sephton will receive another one year of support to continue work on unique roles of FUS (an ALS gene/protein) at the junction where neurons connect, called the synapse. Most of the work done on FUS focuses on its functions in the central region of the neuron called the cell body and in this project, she will examine several critical functions at the synapse that appear to be disrupted by abnormal FUS. When there is a mutation in the FUS gene, it causes ALS and it is important to consider all ways and locations in which this altered FUS can affect motor neuron degeneration. Dr. Sephton will was also a recipient of both an ALS Canada Bridge Grant and an ALS Canada-Brain Canada Career Transition Award in 2015 to work towards these goals and this additional year of funding will provide and opportunity to further advance her work early in her career and provide a bigger foundation of data for future applications.