Research Grants 2024-2025

Applicant
Institution
Title
Award
Dr. Gabriel Bossé
Université Laval
Quebec, QC
Elucidating the long-term neurobiological impact of neonatal opioid exposure
2024/2025: $40,000
2025/2026: $40,000
2026/2027: $40,000
Dr. Mathieu Flamand
Université Laval
Quebec, QC
Unraveling the role of m6A RNA methylation in the pathogenesis of neurodegenerative disease
2024/2025: $40,000
2025/2026: $40,000
2026/2027: $40,000
Dr. Anthony Flamier
CHU Sainte-Justine
Montreal, QC
How to build an attractive cell body – Regulation of subcellular synaptic specificity
2024/2025: $40,000
2025/2026: $40,000
2026/2027: $40,000
Dr. Cairan Murphy-Royal
Centre Hospitalier de l’Université de Montréal (CHUM)
Montreal, QC
Investigating the role of astrocytes in sleep-wake disturbances
2024/2025: $40,000
2025/2026: $40,000
2026/2027: $40,000
Dr. Galen Wright
University of Manitoba
Winnipeg, MB
Long-read RNA sequencing in human models of Rett syndrome
2024/2025: $40,000
2025/2026: $40,000
2026/2027: $40,000
Click on the Research Grant Title to view a brief description of Currently Funded Research Grants below


A Brief Description of Currently Funded Research Grants 2024 – 2025


 

Elucidating the long-term neurobiological impact of neonatal opioid exposure

Dr. Gabriel Bossé
Université Laval
Quebec, QC

Introduction: The widespread usage of opioids as painkillers during pregnancy and the current opioid addiction crisis has led to a sharp increase in the number of newborns exposed to opioids in the womb. A tragic consequence of this epidemic has been an increase of 242%, over the last ten years, of newborns suffering from a syndrome called: Neonatal Opioid Withdrawal Syndrome (NOWS). Newborns with NOWS exhibit symptoms such as difficulty feeding, inconsolability and uncontrolled limb movements, and current treatments involve using opioids. Unfortunately, in humans, prenatal opioid exposure can also lead to long-term defects such as learning difficulties, and anxiety, while in rats, neonatal exposure to opioids leads to delayed learning and impairs attention and impulse control.
At the moment, neonatal opioid exposure’s biological and molecular consequences remain largely unknown, and there are currently no treatment options to prevent the development of long-term effects associated with NOWS. As such, there is an urgent need to develop new approaches to study NOWS and its consequences to develop novel therapies for these patients.
While rodents are most commonly used in neuroscience, novel models such as zebrafish are gaining popularity. These small fish present several advantages, such as a lower cost, the ability to use genetic tools, and the ability to perform large-scale drug discovery approaches. Furthermore, zebrafish are social animals capable of learning small tasks, and they possess a complete nervous system. Their genome shares 70% with humans, and many neuronal pathways are conserved from fish to humans.
Objectives: This research program seeks to understand the underlying molecular mechanisms driving the long-term impact of prenatal opioid exposure on behaviour.
It has been shown that exposing zebrafish embryos to morphine will affect neuronal development and locomotion. Based on these observations, we performed a detailed behavioural characterization of the long-term effects of embryonic morphine exposure.
Our preliminary data demonstrate that this exposure leads to sleeping impairment, increased stress, and changes in thermosensation at the larvae stage. Moreover, at the adult stage, fish exposed to morphine during development were more anxious and less social. Taken together, we observed similar long-term consequences in zebrafish as in other animal models and humans. This validates that zebrafish could be used to study the biological mechanisms driving these changes.

Outline of research: Our behavioural characterization revealed both short-term and long-term behavioural alterations following embryonic morphine exposure. For this proposal, we aim to bridge the gap between molecular and behavioural effects of neonatal morphine exposure. We aim to study the impact of neonatal morphine exposure on gene expression levels and patterns in the brain with an emphasis on important neurotransmitter receptors. Zebrafish larvae are fully transparent, and tools have been developed to measure neuronal activity in live animals. These approaches will allow us to identify the neurobiological impact of neonatal opioid exposure at the molecular level.

Project benefits and application of findings: Taking advantage of the strengths of zebrafish, this research program will lead to the discovery of novel biological pathways implicated in the development of long-term adverse effects after neonatal morphine exposure. This will establish the first program using zebrafish to model NOWS in Canada. Our methods will shed light on biological mechanisms and provide a better understanding of the underlying physical consequences of NOWS. There is currently no cure for these adverse effects, and the pathways and potential drug candidates we will uncover will also provide novel therapeutical options for children.


 

Unraveling the role of m6A RNA methylation in the pathogenesis of neurodegenerative disease

Dr. Mathieu Flamand
Université Laval
Quebec, QC

Introduction: Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are debilitating neurodegenerative diseases characterized by the progressive loss of motor neurons and cognitive decline, respectively. There is an increasingly recognized convergence of clinical, pathological, and genetic aspects between ALS and FTD, with nearly half of ALS patients displaying cognitive impairments. Both diseases are now recognized as different manifestations of a common neurological disease spectrum. However, the precise mechanisms driving the pathogenesis of these diseases remain elusive, and therapeutic options are limited. To develop new treatments and diagnostic tools, it is important to understand the molecular mechanisms involved in these diseases. Common observations in both diseases are the presence of toxic aggregates of the RNA-binding protein TDP-43, as well as changes in RNA regulation, a process crucial to the health and function of neurons. Recent studies have suggested a link between ALS and FTD and the RNA modification N6-methyladenosine (m6A), which itself has important roles in brain function. However, previous studies have revealed conflicting findings, and we do not yet understand the contribution of m6A to these diseases and cognitive impairment in general.

Objectives: Our long-term research aim is to understand the contribution of RNA-binding proteins and RNA modifications in learning and memory formation, and to understand their contribution to neurodegenerative disorders causing dementia. We hypothesize that changes in m6A levels in ALS and FTD contribute to the loss of neuronal activity and cell death, resulting in accelerated neurodegeneration. In this project, our objectives are to characterize the alteration of the m6A pathway in cellular models of ALS and FTD, determine the impact of its dysregulation on global gene expression, to the function of TDP-43, and determine if m6A can be targeted to alleviate neurodegenerative phenotypes.

Outline of research: Towards this, we will follow three specific objectives:
1) Characterize the alteration of the m6A pathway in ALS/FTD: We will use an established cell line and primary mouse neurons to model ALS and FTD in vitro by introducing genetic features associated with familial forms of both diseases. We will next use state-of-the-art genomics methods to profile m6A and measure the abundance of the proteins of its pathway in healthy and diseased cells. Global changes in gene expression will also be assessed and correlated to alterations in m6A. This approach will allow a better understanding of the link between genetic features of these diseases and the levels and function of m6A.
2) Investigate the functional interplay between m6A and TDP-43: We will next investigate how TDP-43, a central player in the pathogenesis of ALS and FTD, is interacting with the m6A pathway in normal and neurodegenerative conditions. Using a unique genomic tool I developed, we will identify the targets of both m6A and TDP-43 and characterize the genetic pathways controlled by both. Their concerted effects will be validated using biochemical tools and molecular biology methodologies. This approach will enable a new understanding of the interplay between two important contributors to RNA regulation in ALS and FTD.
3) Assessing the targetability of the m6A pathway in ALS and FTD: Using genetic tools such as CRISPR/Cas9, we will remove individual genes of the m6A pathway in neurons determine if they can be targeted to prevent neurodegeneration in vitro. Then, using small molecule inhibitors of enzymes responsible for depositing and removing m6A, we will determine if a pharmaceutical approach can be used to alleviate the loss of neuronal function and cell death observed in ALS and FTD patients.

Project benefits: This project proposes original and innovative studies on m6A in ALS and FTD. It will enable a unique understanding of the disruption in the m6A pathway in neurological disorders and reveal its contribution to changes in gene expression in concert with TDP-43 during neurodegeneration. This project will not only address gaps in our understanding of m6A dysregulation in ALS and FTD but will also lay the foundation for identifying potential biomarkers and therapeutic targets in these severe diseases.


 

Unraveling the impact of proteasome dysfunction on epigenetic regulation and neurodevelopment.

Dr. Anthony Flamier
CHU Sainte-Justine
Montreal, QC

Introduction: Understanding the complexities of brain development and the genesis of neurodevelopmental disorders remains a major challenge in neuroscience. Neurodevelopmental disorders, such as intellectual disabilities and autism, profoundly impact cognitive function, yet their underlying biological mechanisms are not fully understood. This proposal focuses on unraveling the role of the UBR7 gene, which recent studies suggest is crucial for normal brain development and function.
Objective: The primary objective of this research is to elucidate how deficiencies in the UBR7 gene contribute to changes in the brain’s epigenetic landscape that may lead to neurodevelopmental disorders. By exploring the intricate mechanisms through which UBR7 regulates brain development, this study aims to lay the groundwork for potential therapeutic strategies that could mitigate or even prevent these disorders.
Outline of Research: Our research utilizes two innovative models: in vitro cerebral organoids derived from induced pluripotent stem cells (iPSCs) and an in vivo mouse model. The iPSCs are engineered from cells of individuals with a diagnosed UBR7 deficiency, allowing us to observe the developmental trajectory in a controlled environment that mimics human brain development.
1-In Vitro Model: We will generate cerebral organoids from UBR7-deficient iPSCs to simulate brain development. These organoids will be analyzed over several months to track epigenetic changes, such as DNA methylation and histone modification, which influence gene expression critical for brain development.
2-In Vivo Model: Parallel to our in vitro work, we will study mice genetically modified to lack UBR7. These mice will help us understand the physiological and behavioral outcomes of UBR7 deficiency, providing a comprehensive picture of its role in neurodevelopment.
Projected Benefits and Application of Findings: The findings from this study are expected to have significant implications for the understanding of neurodevelopmental disorders at a molecular level. By identifying the specific epigenetic mechanisms altered by UBR7 deficiency, this research could lead to novel diagnostic markers for early detection of such disorders. Furthermore, understanding these pathways opens the possibility of developing targeted therapies that could correct or compensate for these epigenetic abnormalities. Ultimately, this research aims to provide new insights into the prevention and treatment of disorders like intellectual disability and autism, significantly improving the quality of life for affected individuals and their families.


 

Investigating the role of astrocytes in sleep-wake disturbances

Dr. Cairan Murphy-Royal
Centre Hospitalier de l’Université de Montréal (CHUM)
Montreal, QC

Introduction: Early-life stress in the form of neglect, abuse, and malnutrition during childhood has been shown to result in higher incidence of psychiatric disorders and cognitive dysfunction later in life. This human condition is faithfully reflected in rodent models using adverse experiences including maternal separation and deprivation, resulting in complex behavioral dysfunctions. Consider that sleep disturbances are a common and problematic symptom of psychiatric disorders, with both insomnia and hypersomnia forming hallmark symptoms of various stress disorders, understanding the biological mechanisms holds the potential to have benefits across a wide number brain disorders.

Objectives: Our project sets out to determine how stress affects the lateral hypothalamic nucleus of the brain. This small but important brain region is critical for regulating wakefulness. Experimental manipulation of the lateral hypothalamus has been shown to induce narcolepsy, when activity is decreased, and promote insomnia when this brain region is hyperactive. Considering the vast number of brain disorders associated with poor sleep, whether that be sleep quantity, quality, or disrupted sleep patterns, we aim to further understand the role of the lateral hypothalamus in regulating sleep and how it is affected by stress.

Outline of Research: The unique aspect of our project lies in our preliminary observations showing that a type of brain cell, called the astrocyte, is critical for regulating activity of neurons in the lateral hypothalamus. Astrocytes in this particular brain region remain critically understudied. As such, we want to dissect the role of astrocytes, how they are affected by stress, and whether dysfunction of these brain cells results in sleep-wake dysfunction.
To address this we will carry out three primary aims 1) investigate the effects of early-life stress on astrocyte structure and function, 2) determine the downstream consequences for wake-promoting orexin neurons, and 3) identify whether targeting astrocytes improves sleep-wake cycles.

Projected Benefits: Our work will identify whether astrocytes might represent a novel therapeutic avenue to treat stress-induced sleep disorders. As sleep is key to proper brain function, including learning, memory, cognitive function, understanding this essential mechanism is key to brain and cognitive health. While our project uses stress as a tool to disrupt sleep and understand the neurobiological mechanisms, the data and methodologies outlined in our proposal could be readily applied to a wide variety of disorders of the mind that present sleep dysfunction as a symptom. This can include children with autism that often report difficulty to fall asleep with frequent waking during the night. If we consider that the lateral hypothalamus is a wake-promoting brain region, future studies could determine causal links.


 

Long-read RNA sequencing in human models of Rett syndrome

Dr. Galen Wright
University of Manitoba
Winnipeg, MB

Introduction: Rett syndrome (RTT) is a rare neurodevelopmental disorder that affects mainly girls. Symptoms appear in children between 6 and 18 months old and include loss of movement and communication skills, slowed growth, and breathing difficulties. Over 95% of affected individuals have a mutation in the MECP2 gene, which plays a key role in brain development. Understanding how this gene mutation affects brain development and related pathways in RTT can offer insights into other neurodevelopmental disorders.

Human pluripotent stem cells (hPSCs) offer a promising experimental system for studying the underlying disease biology of RTT and other related neurological disorders, which can help identify new treatments. Stem cells can be converted into various cell types, including neural cells, allowing scientists to observe the impacts of MECP2 mutations in a controlled manner. Notably, new methodologies have been developed to use these cells to create miniature models of the human brain called organoids. Since hPSC models can mimic human biology, we can use cutting-edge genomic technologies to generate a wealth of information to help us understand the processes that occur during disease.

Projected Benefits and Applications of Findings: This research will help us understand how RTT and other neurodevelopmental disorders affect our brains at both the molecular and cellular levels during the early stages of development. By using new genomic technologies, such as long-read RNA sequencing, we will be able to precisely capture disease-associated changes in how genes are used in individual brain cell types. This will allow us to accurately study these disease-relevant changes at scale for the first time. Understanding the specific gene expression changes and isoform alterations in neural cell types can help identify possible therapeutic targets for these conditions. In the long term, the knowledge gained from this research could inform the development of new treatments for RTT and related disorders, ultimately improving the quality of life for affected individuals and their families. Finally, this research will train students and personnel in innovative human genomics and stem cell-based methodologies, with an emphasis on neurodevelopmental disorders, therefore enhancing Canadian research capacity in these areas.

Objective: The primary objective of this research is to use long-read RNA sequencing in human stem cell models of RTT to study disease-related changes in gene expression in different brain cell types. Long-read RNA sequencing is a new genomic method that allows us to identify the exact versions of individual genes expressed in individual cell types. Rather than piecing together incomplete fragments from genes, as is used in traditional approaches (e.g., short-read sequencing), long-read technologies allow us to observe complete versions of expressed genes directly. These technologies have only recently become cost-effective, which now allows us to study how the loss of MECP2 leads to RTT more accurately than was previously possible.

Outline of Research: We will employ a human model of RTT created from hPSCs using both two-dimensional (2D) layers of brain cells and three-dimensional (3D) brain organoid models. Aim 1 will differentiate hPSCs into both disease-relevant neural progenitor cells and neurons and use bulk long-read RNA sequencing to comprehensively study these cells in detail. This approach will allow us to identify whether common and rare versions of genes in these cells are altered in RTT. Aim 2 will generate 3D brain organoids from RTT hPSCs which will allow us to study how several brain cell types are affected in disease at the gene expression level. We will combine cutting-edge approaches – single-cell sequencing combined with long-read sequencing – to study whether different versions of genes are used more or less frequently in individual brain cell types in disease. These complementary approaches will allow us to comprehensively analyze how the loss of MECP2 impacts gene regulation and human brain development at a resolution that was not previously possible.