LSI researchers awarded funding in Fall 2024 CIHR Project Grant competition

Congratulations to all LSI researchers awarded funds in the Fall 2024 CIHR Project Grant competition! A description of the funded projects is provided below. In total, these grants were awarded >$7.7 million from this competition.

Epilepsy, a brain disorder characterized by recurrent seizures, imposes significant challenges for individuals and society. Many individuals with epilepsy exhibit “pharmacoresistance”; that is, seizures that fail to respond to medication. Revealing the causal neurobiology underlying epilepsy – for example, neuron types and molecules that drive seizures – will likely prove critical for identifying next-generation for targets and treatments to overcome this pharmacoresistance. In our specialized collaboration spanning researchers in experimental neuroscience, along with clinicians involved in epilepsy neurosurgery and neuropathology, we seek to develop this knowledge on neurons and molecules that cause seizures. Through this collaboration, we have discovered an atypical neuron type, present in both the mouse and human brain, which may act as a seizure “detonator” due to its heightened excitability and extensive long-range connections. We will study this neuron type using a complement of experiments in mice, as well as living human brain tissue obtained from informed, consenting participants undergoing brain surgery for epilepsy. To begin, in epileptic mice, we will measure the ability of detonator neurons to drive widespread neural activity and seizures. In living human brain tissue, we will assess whether detonator neurons are similarly capable of driving widespread neural activity in the human brain. Using cutting-edge spatial transcriptomics experiments in both mice and humans, we will identify druggable molecules in detonator neurons and other cell types, and use this knowledge to disrupt these molecules using novel drug and genetic interventions. In the long term, these results may inform and guide the generation of new therapeutic approaches for epilepsy that was previously deemed pharmacoresistant.

This research addresses critical challenges associated with blood transfusion: the prevention of bacterial growth and quality of stored blood. Platelets and whole blood are given to patients suffering hemorrhage, cancer, bleeding and other disorders. Currently platelets are kept in bags made of plastics not originally designed for cell storage. Fresh whole blood (FWB) is collected and stored in similar bags. Current bag materials have no antibacterial properties, with one in every 1000-1500 platelet units being contaminated by bacteria, rendering the unit highly hazardous to recipients. FWB is used within 24 h of collection due to fear of infection. Canada has introduced pathogen-inactivating systems to combat these bacteria, but the process damages the platelets and it has been shown that bacteria can escape inactivation causing harm. There is no technology currently available to present bacterial growth in whole blood or any red cell containing transfusion products. This research addresses this critical challenge, by developing self-sterilizing devices for the collection, processing and storage of the blood, and to ensure the blood remain bacteria-free while improving their shelf-life and quality for transfusion. We will achieve this goal by leveraging cutting-edge antibacterial coatings developed by our lab. We will screen various coating compositions to identify top performers against a broad spectrum of clinically relevant bacteria. We will also develop novel surface attached antimicrobial agents identified using our novel screening method. These novel coatings easily applied to blood storage bags will be optimized for long-term antibacterial function, and to ensure they stabilize the platelets and whole blood. Through this technology, we aim to eliminate the fear of infection in transfusion products to minimize the adverse effects associated with transfusions, and improve the safety, decrease shortage and increase their shelf-life.

Our knowledge of the functions for membrane contact sites within cells is extremely limited yet they are present in all eukaryotic cells, from budding yeast to humans. Hence employing models such as the budding yeast to make breakthrough fundamental discoveries will lead to a better understanding of membrane contact site function in humans and their roles in disease. A major contributing organelle to membrane contact sites is the endoplasmic reticulum, which makes contacts with all other organelles in the cell. Some well characterized contacts are with mitochondria, plasma membrane, lysosomes, peroxisomes, Golgi and endosomes. However, membrane contact sites are not restricted to the endoplasmic reticulum and have been observed between mitochondria and plasma membrane and vacuoles/lysosomes. Main functions ascribed to membrane contact sites so far are in lipid and calcium traffic, cell signaling, and organelle biogenesis/homeostasis. The work proposed in this grant will use yeast to identify new components of membrane contact sites and define their functions and organization in much more detail. This will be through the use of high-resolution microscopy, genetic and biochemical approaches. The importance of membrane contact sites in human disease is also being uncovered through defining proteins with functions at membrane contact sites and include cancer, obesity, diabetes, and neurological diseases. However, many of the disease mechanisms have yet to be defined largely due to a lack of a clear understanding of the physiological functions of MCSs. This work will expand our understanding of these mechanisms.

Blood transfusion is a life-saving therapy used to treat a variety of medical conditions. In some situations, such as trauma or surgery, the transfused blood is only needed for a short period of time (less than two days). In other situations, such as for patients with certain hereditary diseases or patients undergoing certain chemotherapies, the transfused blood is needed for a long period of time (several weeks). For this second group of patients, it is important for the transfused blood to last as long as possible in order to reduce the need for repeated transfusions and the potential complications associated with blood transfusions. The longevity of transfused blood depends on the donor, but currently, there are no methods for predicting which donor can provide long-lasting blood. Our study will investigate the potential to estimate the longevity of donor blood from the deformability, or softness, of individual red blood cells. Red blood cells must traverse the entire circulatory system every minute. During this journey, they must be repeatedly deformed to transit through the microscopic capillaries in the body. The loss of red blood cell deformability is associated with the removal these cells from circulation. Therefore, red blood cell deformability is a likely predictor of donor blood longevity in transfusion recipients. If successful, our study can provide an approach to identify donors who can provide long-lasting blood. Blood from these donors could then be reserved for patients that could benefit from long-lasting blood in order to reduce the number of transfusions and transfusion-associated complications.

Mycobacteria, such as Mycobacterium tuberculosis, are the deadliest bacterial pathogens, responsible for over 1.5 million deaths annually. One of the major reasons for the success of these pathogens is their unique and highly impermeable cell envelope, consisting of two membranes. While this envelope serves as a crucial barrier for survival during infection by conferring resistance to numerous antibiotics, it must simultaneously permit nutrient uptake and protein secretion for vital interactions with the host. To overcome these challenges, Mycobacteria have evolved a unique secretion system, the type VII secretion system (T7SS), as their main machinery that allows them to transport substrates, while maintaining the integrity of their cell envelope. While some substrates are important for virulence, others are essential for the uptake of specific metabolites that support mycobacterial growth. Elucidating the mechanism of T7SS secretion and the role of individual effector proteins is therefore crucial for understanding the success of pathogenic mycobacteria. Outcomes from this work could facilitate the identification of novel targets for drug and vaccine development.

The stress hormone cortisol (aka hydrocortisone) is a glucocorticoid that is needed for development and function of many body systems, and is one of the most widely-prescribed medications. In our blood, cortisol is bound to a protein called corticosteroid-binding globulin (CBG) which carries it in the circulation and controls how it works on target tissues. When the blood levels of CBG are too low, there can be health problems, including poor ability to cope with stress, injury or inflammation, or chronic fatigue/pain and depression. Many of these traits are glucocorticoid dependent, but it remains unclear why these are manifested differently in males and females. A genetically-modified, CBG-deficient rat has been generated to test the role of CBG in modulating sex differences in immune, hormonal, and behavioural responses to stress.

Transposable elements (TEs) are selfish genes that copy themselves and jump around the genome. Sophisticated defense mechanisms prevent TEs from rampant replications and insertions causing devastating mutations that destroy genes, generate breaks in the DNA, and induce chromosomal rearrangements. TE regulation is therefore pivotal to the maintenance of genome stability, and failure to suppress them are associated with myriads of disease phenotypes and illness including tumorigenesis and sterility. Despite the robust and conserved defense mechanisms, TEs repeatedly developed means of evading control. This is evident from their overwhelming abundance across eukaryotes, including the human genome of which they occupy nearly 50%. Using the powerful model genetic organism, the Drosophila, this proposal aims to dissect the molecular bases for two strategies in which TEs adopt during a critical juncture of early embryonic development called zygotic genome activation. The first strategy to be elucidated is the timely activation of TEs during this critical time point allowing them to proliferate before silencing mechanisms are established. The second is for TEs to concentrate their activity at favorable locations to increase their chance to insert into the germline necessary for transmission to the next generation. To understand the mutational and disruptive impacts of individual insertions, we further aim to artificially induce TE insertions through multiple generations. Large numbers of novel TE insertions will allow exhaustive investigations of how individual insertions affect local epigenetic state, gene regulation, and chromatin architectures genome-wide. Understanding how TEs proliferate and how they disrupt genome architecture will critically inform preventative and treatment plans for the plethora of diseases caused by genome instability.

Autophagy is an essential “junk removal” system used by all cells in our bodies to package large objects such as cellular organelles and protein aggregates and deliver these packages to the “cellular incinerator” the lysosome for breakdown. Malfunction of autophagy affects the normal balance between production and breakdown and has been shown to lead to a wide range of human diseases from neurodegeneration, cancer, to infectious diseases. To determine how autophagy is disrupted in the disease state, our laboratory is trying to obtain a comprehensive understanding of how autophagy operates at the molecular level by studying the dedicated protein machinery that coordinates this multi-step process. This project grant application focuses on three components of this machinery (EPG5, HOPS complex, Rab3GAP1/2 complex). Mutations to EPG5, HOPS, and Rab3GAP1/2 lead to the congenital disorders Vici syndrome, HOPS-associated dystonia, and Warburg micro syndrome, respectively. We will investigate the molecular mechanisms of how these proteins and protein complexes check the identities of the junk packages and facilitate their delivery to the lysosome for breakdown. Our proposed research will fill key knowledge gaps in our knowledge on how cells control and execute autophagy degradation essential to health. In the long term, our research will pave the way for the development of autophagy-targeted therapeutics for treating different human diseases, and improve the quality of life of the large number of Canadians suffering from these disorders.

Malaria is a severe illness that impacts millions of people annually, especially in regions like sub-Saharan Africa, Southeast Asia, and parts of South America. In 2022, malaria resulted in 249 million cases and 608,000 deaths worldwide, with young children and pregnant women being the most affected. Although there are vaccines like RTS,S/AS01 available, their effectiveness decreases over time, highlighting the urgent need for better treatments. Our project, partially supported by Eyam Vaccines and Immunotherapeutics Ltd. and the Bill & Melinda Gates Foundation, aims to address this issue with an innovative approach. Instead of producing malaria-fighting antibodies in a lab and injecting them into patients, which is technically challenging and expensive, we are developing a technology that allows the body to create these antibodies on its own through advanced genetic techniques. This technology, which can be applied to malaria as well as other diseases and ailments, is known as the Gemini platform. It is designed to induce strong and long-lasting protection, reducing the need for frequent treatments. It is relatively cheap to produce; it can be freeze-dried and stored without refrigeration, making it ideal for use in areas with limited access to cold storage-something crucial in malaria-endemic regions with scarce resources. Our primary goals are to develop and test this new treatment to determine if it can effectively protect against malaria in laboratory and animal studies. If successful, this technology could greatly enhance and democratize access to more affordable malaria treatments. Looking ahead, this approach could also be applied to other diseases, offering a new way to improve global health. Ultimately, our aim is to reduce the global burden of malaria and other diseases by providing more durable, cost-effective, and accessible solutions.

The COVID-19 pandemic and Mpox epidemics have underscored the need for innovative vaccines to enhance public health responses. The currently approved Mpox vaccine in Canada, JYNNEOS (aka Imvamune), is a live attenuated virus vaccine based on a modified Vaccinia virus. While generally safe, JYNNEOS can cause adverse effects such as myocarditis and complications in individuals with skin disorders. The estimated rate of protection against clade IIb provided by existing vaccines is between 60 and 70%, which might be insufficient to prevent spread, particularly in immunocompromised individuals. Therefore, we are developing and alternative, the first made-in-Canada recombinant Mpox vaccine designed to offer superior safety and immune protection. Our approach utilizes the Gemini platform, a cutting-edge nucleic acid vector, superior to existing mRNA strategies. Gemini is not a virus, reducing potential risks and enabling rapid, cost-effective manufacturing. Our vaccine further uses information from the Jennerator, which is a revolutionary computer-aided design tool that accurately vaccine payloads. Importantly, our vaccine does not require lipid nanoparticles, which enhances its manufacturability and affordability. To ensure the efficacy and safety of our vaccine, we will conduct head-to-head trials comparing our vaccine directly with JYNNEOS, to provide critical data on relative safety, immune response, and effectiveness. Through our partnership with Eyam Vaccines and Immunotherapeutics, who have licensed the technology from the University of British Columbia, we have a robust plan to accelerate the vaccine’s journey from development to clinical trials and eventual commercialization. This proposal aims to advance and validate our innovative vaccine platforms, addressing the urgent need for effective Mpox vaccines and enhancing Canada’s preparedness for future pandemics.

Our research group established unique methods for simultaneously testing thousands of drugs to identify drugs that might be useful for beta cell protection in diabetes. One of these screens was designed to identify drugs that can protect beta cells from immune attack in type 1 diabetes. The drugs we tested were selected from a library of FDA-approved drugs, meaning that they were already optimized for use in people. We identified a drug called carbamazepine that reduced beta cell death in a type 1 diabetes context. Carbamazepine is an interesting lead compound because it works by inhibiting a specific type of sodium channels, encoded by the Scn9a gene, but only when they are over-active. This was the first implication of sodium channels in beta cell survival and opened a whole new area for type 1 diabetes drug development. Importantly, pancreatic beta cells have a unique complement of sodium channels, meaning that this drug and its derivatives are likely to be relatively beta cell-selective. We validated our cell culture data by showing that carbamazepine reduces diabetes incidence in the gold standard NOD mouse model. We also determined that carbamazepine can protect human beta cells in the lab. In the proposed studies, we plan to continue this exciting line of investigation towards a new diabetes drug and answer several key questions that arose from our previous studies. First, we need to understand exactly how carbamazepine and Scn9a affect the activity of the beta cells. Second, we must determine how carbamazepine and Scn9a protect beta cells in the context of type 1 diabetes. Third, we need to study how beta cell sodium channels may modulate local immunity to slow type 1 diabetes. These studies will help us determine whether beta cell sodium channels are a viable therapeutic target in type 1 diabetes.