Multifunctional immunomodulating conjugates for targeting and treating glycocalyx dysfunction in inflammatory conditions

Diseases that involve the heart or blood vessels, autoimmune diseases (e.g. diabetes, multiple sclerosis), or the rejection of transplanted organs affect about 1 in 3 Canadians and constitute a significant cost to the Canadian economy. In the perpetuation of these diseases glycocalyx shedding plays a key role. The glycocalyx (literally meaning “sugar coat”) is a sugar polymer-based structure that covers the surface of the cells, which are lining all organs and blood vessels. It lies at the interface between bloodstream and organ tissue and represents the protective front line against inflammatory and immune-mediated diseases. Thus, we aim to specifically target and treat glycocalyx dysfunction by rapidly rebuilding it through a new cell surface engineering approach, which should enable organs to maintain or reestablish their function. To do so, we will develop polymer conjugates which can selectively bind and retain on the endothelial cell surface. The conjugates will present sugar moieties which resemble the natural glycocalyx layer. We anticipate to realize a novel approach with significant therapeutic potential to improve treatment for diverse disease conditions where glycocalyx dysfunction is contributing to the pathology.

T-cell repertoire analysis for immune monitoring in renal transplantation

Kidney disease affects 1 in 10 Canadians with an estimated cost of over $2 billion per year. Transplantation is the treatment of choice for kidney failure, but unfortunately approximately 30% of kidney transplants are lost to severe immune rejection. This leads to approximately 500 Canadians losing their transplant every year and returning to dialysis. These patients have a four-fold increased risk of death, decreased quality-of-life, and a cost of up to $1 million each to the healthcare system over their remaining life. Despite improvements in transplant care, there are still no proven methods to detect early immune rejection. Our goal is to develop a new minimally invasive blood-based test to monitor the immune system of transplant patients to detect immune rejection before kidney damage happens. This would allow transplant doctors to intervene early with powerful immune regulating medications and prevent irreversible damage to the transplant kidney. Our approach would not only benefit patients and their families with improvement in survival, quality of life, caregiver burden, and personal health expenses, but also the healthcare system, with reduced costs related to dialysis, re-transplantation, and improved organ availability.

The effect of temperature on brain bioenergetic stress in hypoxia

Cooling the brain is a therapeutic strategy to protect it from stress. The long-held belief is that cooling the brain reduces its activity — and thus its need for oxygen — thereby tilting a favourable balance of oxygen supply and demand. However, recent data from our lab challenges this paradigm. We have shown that brain blood flow is reduced by whole-body cooling, and this dramatically impairs oxygen supply to the brain. Therefore, it is important to know exactly how much the brain’s activity is reduced so that we can determine whether the balance of oxygen supply and demand is improved or further disrupted. Surprisingly, this is unknown in the human brain. Our objective is determine how the brain’s oxygen supply and demand is affected by cooling and heating, and how this impacts its resilience to stress. We will heat and cool healthy human subjects and expose them to low oxygen, whilst measuring markers of brain stress. We will then collect the same markers of brain stress in patients with brain injury before, during and after therapeutic cooling. Together, these studies will expose how temperature affects the brain’s resilience to stress and provide rationale for how best to harness the cold to protect the brain.

Impact of prostate cancer subtypes on bone remodeling and microarchitecture in metastatic lesions of the spine: A combined cellular, genetic and mineralization study

One in nine men will develop prostate cancer (PC) in their lifetime. Although modern therapies have increased the survival rate, almost all advanced cases will metastasize to bone, with the axial skeleton being the most frequent location. Bone metastases (BM) are the most severe complications of PC generating severe pain, fractures, and spinal cord compression. So far, it is not clear how PC BM are related to pain and fracture. Most cancers that generate bone complications, are associated with bone loss. However, PC is associated with bone formation. The aims of this project are to understand the structure of this new formed bone, how prostate cancer cells induce these changes, and if there are any specific types of PC associated with these changes. The ultimate goal is improving disease management and preventing complications of PC BM.
I have observed the microstructure structure of PC BM in mineral and protein content. Also, I have identified different types of PC cells in the PC BM, meaning the cells are undergoing a transformation process in the bone. These results are unprecedented, and my aim is now to expand the sample size and to explore the structure of PC BM in greater detail in order to prevent its severe consequences.

Programming bordetella pertussis to produce novel vaccines

Pertussis (whooping cough) continues to be a problem despite high vaccination coverage against Bordetella pertussis, the bacterium that causes the disease. Annually, there are 24 million cases of pertussis and ~160,700 deaths worldwide. Pertussis is a respiratory disease that is transmitted from person to person through airborne droplets and poses a threat to unvaccinated infants and children whose immunity has dropped. Currently, there are two forms of the vaccine in use. The first is the killed whole-cell vaccine (wP), which is effective, but has side-effects such as swelling at the site of injection and fever. These adverse effects have diminished its acceptance in high-income countries and led to its replacement by the acellular vaccine (aP) that only contains purified components of the organism. While the aP vaccine protects against getting pertussis, it does not prevent transmission of the disease and fails to provide long-term immunity.
We aim to develop two new vaccine candidates: a revised wP and a novel aP to control the re-emergence of pertussis. This will be done through modifying some of the structural components of the bacteria to either alleviate the side effects or overcome the deficiencies of the wP and aP vaccines.

Endothelial calcium dynamics regulating cerebrovascular function and capillary stalling in the healthy and diabetic brain

The brain is a metabolically demanding organ . Mismatch between blood flow and demand (from neurons) leads to a disruption and in extreme cases injury. Because the smallest blood vessels in the brain are narrow, they are prone to becoming obstructed by circulating cells and debris. This is exacerbated in Diabetics, with “sticky” blood vessels. The cells of blood vessels, endothelial cells, are more than just “pipes”, they form large physically connected networks between themselves. An important regulator of these networks, and a signal to communicate between them, is waves of calcium flowing into cells, which can propagate between these cells. How Diabetes affects these networks of blood vessels, and in turn impact the health of the brain is unknown. Thanks to new genetic tools with state of the art microscopes, we can directly observe these calcium fluxes into endothelial cells in the living, awake, mouse brain, and especially when these blood vessels become occluded. Combined with simultaneous monitoring of blood flow and neural activity I will be able to directly measure concurrent changes in brain activity, blood flow and calcium fluxes to investigate these dynamics in the living healthy or Diabetic mammalian brain.

The role of mechanics in EMT and cancer metastasis

The World Health Organization reports that cancer is the second leading cause of death globally, responsible for 1 in every 6 deaths. This ratio doubles in Canada, with the Canadian Cancer Society estimating that nearly 1 in 2 Canadians will develop cancer, and about 1 out of 4 will die from it.

Recent anticancer therapies target the epithelial-to-mesenchymal transition (EMT), a process that converts tightly bound cells into loosely associated motile cells. In cancers, this results in progression with metastasis and improved resistance to treatments.

Evidence shows the role of mechanics in driving EMT but how the biochemistry and the mechanics coregulate this process remains largely unknown.

We propose to investigate this question in the case study of stem cell cultures, which undergo EMT in a controlled environment. We will develop a mathematical model to link mechanical stresses and cytoskeletal energetics, and we will validate it experimentally in collaboration with the Zandstra Lab.

This proposal will enhance BC’s and Canada’s leadership in healthcare-oriented research, as understanding EMT is essential not only for cancer but also for many other biological processes, such as organogenesis and tissue regeneration.

Synthetic feedback control of TCR signaling to guide T cell development in vitro

T cells are an important component of our body’s adaptive immune system, helping to identify and overcome diverse diseases. An emerging treatment for cancer, viral infections, and other diseases is to engineer patient’s T cells to recognize and respond to diseased cells. However, because of the reliance on patient-derived T cells, such treatments are highly expensive. To lower costs and increase accessibility to T cell therapies, our laboratory is developing methods to generate T cells from an unlimited and readily-available source: human pluripotent stem cells. Pluripotent stem cells give rise to every cell in our bodies, including T cells, and can be grown indefinitely in laboratory settings. Our current process for producing T cells from stem cells has made great progress, but lacks control over key parameters such as whether the T cells will become “helper” cells that stimulate the immune system or “cytotoxic” cells that directly kill diseased cells, and if they will provide long-term memory or have strong, short-term effects. In this project, I will genetically engineer stem cells such that we can produce T cells with these diverse properties on-demand, thereby enabling the next generation of off-the-shelf T cell therapies.

Engineering Platelets using therapeutic mRNA

Platelet cells are routinely transfused during treatment of a range of conditions, due to their specialized roles in hemostasis. Despite the significant potential to enhance the efficacy and applicability of platelet transfusions, no techniques have yet been developed to engineer modified platelets. mRNA therapeutics is a promising novel class of nanomedicine with broad clinical applicability, capable of enhancing the physiological function of target cells by modifying cellular protein expression. The therapeutic potential of mRNA editing is particularly relevant to transfusion science, where the mechanisms of delivery to patients are well established. By engineering platelets using gold standard mRNA transfection strategies, their therapeutic potential can be maximized for diverse applications.

Engineered platelets will be created using cutting-edge mRNA lipid nanoparticles. Successful mRNA editing will create platelets with enhanced biochemistry and improved hemostatic function. Results generated from this project will address knowledge gaps in platelet translation mechanism, and guide forthcoming research on the next generation of blood products, improving current standards of care in blood transfusion.

Mapping chronic social isolation-induced brain activation in mice with machine learning-based phenotyping of behavioral deficits to pilot translational assessment of psychomotor disturbance

Loneliness is becoming increasingly recognized as a serious threat to mental health. Social isolation is detrimental to adult brain function and behavior across mammalian species. Chronic social isolation in rodents has been found to lead to depression-, anxiety-, and psychosis-like behaviors as well as signs of abnormal locomotor habituation, fear responses and aggression. However, our understanding of how and why social isolation is risky for health — or conversely — how and why social ties and relationships are protective of health, remains quite limited. Our lab makes use of advanced brain imaging and recording techniques to map connections between brain areas. We plan to use these techniques to help us to first understand the neuropsychiatric basis of chronic social isolation in animal models. A machine learning algorithm will be used to classify large behavior datasets automatically and objectively, and potentially uncover new pathological behavioral patterns that have been overlooked by human observers. Mapping large-scale brain functional connectivity associated with social isolation–induced behavioral deficits may shed light on the etiopathogenesis of mental disorders and lead to the identification of therapeutic targets.