Projects Supported by the netCAD Blood4Research Program

Project ID: 2018.014b4r

Project Approval Date: 2018-03-29

City: Vancouver

Use of Animals: No

Study Approval Date: 3/29/2018

Canadian Blood Services Mandate: Transplantation medicine

Summary: Blood and marrow transplantation (BMT) is the only established immune therapy for blood cancers. While its effectiveness has improved significantly with increased safety, it is still severely limited by graft vs host disease (GvHD). GvHD is the rejection of not only the leukemia cells, which is the goal, but also of normal tissues in the body.  The chronic form of GvHD, cGvHD, can last for years and causes both major physical and health limitations, but also can lead to shortened life span.  Thus, new strategies to minimize cGvHD, but keep the anti-leukemia effects of BMT, are critical. Recently, our group observed that in a group of bone marrow donors from national are important in inhibiting cGVHD. There are several NKreg populations that have been associated with immune tolerance and at this time we are uncertain which of these populations inhibits cGVHD. We hypothesize that we can develop cellular therapy for patients receiving a BMT to decrease the development of cGVHD. In this proposal, we aim to optimize in vitro propagation of NKreg population. This will be a critical building block toward developing NKreg therapy to minimize or prevent cGvHD in BMT.  As there are presently no known NKreg trials, this would proceed toward the first such trial in Canada and possibly worldwide. In this project no genetic analysis and/or stem cell research is proposed.

Project ID: 2018.018b4r

Project Approval Date: 2018-05-17

City: Toronto

Use of Animals: Yes

Canadian Blood Services Mandate: Transplantation medicine

Summary: For the patients who need the organ or therapeutic cell transplantation, immune rejection following the transplantations means the host immune system recognizes the transplant as non- self and start to attack the transplant. Immune rejection threatens patients’ life and in clinics, immunosuppressive drugs can help inhibit the immune system and increase the transplant survival rate. However, there are many long-term side effects such as toxicity of immunosuppressive drugs, higher infection risk and graft versus host disease (GVHD). We aim to explore how to induce the local immune tolerance and recipient to protect donor’s organ/tissues/cells after organ transplantation for better survival and life quality without immune suppressive drugs. Considering the ability of human pluripotent stem cells (hPSCs) to differentiate into different mature lineages, such as retinal pigment epitheliums (RPE). our approach is to genetically modify human pluripotent stem cells and screen the candidate immunomodulatory genes that have potential functionalities to regulate the immune rejection to non-self organs/tissues/cells. 

Project ID: 2018.020b4r

Project Approval Date: 2018-06-08

City: Toronto

Use of Animals: No

Canadian Blood Services Mandate: Does not benefit Canadian Blood Services

Summary: Immune cells such as macrophages are known to play a critical role in modulating the tumour microenvironment and in determining how the disease progresses. They display a spectrum of behaviors: some promote tumour growth while others attack tumor cells. The balance of behaviors exhibited by immune cells within tumours is therefore critical in determining whether tumour cells are attacked by the immune system, or whether the tumour microenvironment is reprogrammed to promote tumour growth, therapy resistance and disease progression. The effectiveness of current immunotherapy approaches that attempt to enhance the balance of anti-tumour immune cells is limited to a number of cancer types and vary significantly within the patient population. This is in part because the mechanisms of how immune cell interactions with tumour cells are modulated to an anti-versus pro-tumourigenic state within the complex tumour microenvironment are poorly understood. Specifically, this project will probe macrophage-tumour interactions in the tumour microenvironment to understand how to develop more effective immunotherapies and potentially augment response to standard-of-care therapies.

Project ID: 2018.028b4r

Project Approval Date: 2018-07-05

City: Ottawa

Use of Animals: Yes

Canadian Blood Services Mandate: Does not benefit Canadian Blood Services

Summary: Our research focuses on understanding how fat turnover is regulated by autophagy. This is an emerging field of study, especially in the context of heart disease. Our research combines new scientific methodologies to address major gaps in our knowledge and to provide fundamental insights on how lipophagy regulates fat breakdown and inflammation in plaques. Lipophagy is a recently discovered cellular process, but the molecular components of this pathway are unknown. Identifying the lipophagy machinery and defining the role of lipophagy in atherosclerosis is the first step toward designing new therapeutic tools to promote reverse cholesterol transport from the artery wall. For this, we purify low density lipoproteins (LDL) from human plasma and modify these to generate foam cells, mimicking the lipid-loaded cells that aberrantly accumulate in the vascular wall during atherosclerosis, the major underlying cause of heart disease. Therefore, we are employing human LDL to generate these foam cells in a dish in order to study the mechanisms of cholesterol removal from these cells. Researchers and pharmaceutical companies have been trying to develop drugs to increase reverse cholesterol transport for decades. As such, lipophagy would represent an extremely promising therapeutic strategy to treat heart disease. Simultaneous targeting of fatty deposit breakdown and dampening plaque inflammation would represent an important breakthrough for the treatment of cardiovascular diseases.

Project ID: 2018.033b4r

Project Approval Date: 2018-07-31 

City: Vancouver

Use of Animals: No

Canadian Blood Services Mandate: Transplantation medicine

Summary: Transplanted organs always face the threat of rejection by the host’s immune system.  It is  well  known  that damage  to  transplanted  organs  during  harvest,  transport,  and  implantation  are  risk  factors  for  rejection.  What is not  known  is  how  this  damage  itself  helps  to  sensitize  the  immune  system  to  make  it  more  likely  to  reject  the organs. Our lab studies how “danger signals” produced by damaged cells and tissues stimulate the immune system.  Using human  cells  derived  from  blood  donors,  we  have  found  that  signals  released  from  damaged  cells increase  the  ability  of  these  cells  to  trigger lasting  immune  responses.  We propose  to  study  this  phenomenon further using human cells as well as drugs linked to human serum proteins to stimulate immune responses.

Project ID: 2018.038b4r

Project Approval Date: 2018-08-30

City: Edmonton

Use of Animals: No

Canadian Blood Services Mandate: Transfusion medicine

Summary: We want to determine the blood compatibility of in-house prepared nanoparticles that are being investigated for gene therapy. The nanoparticles will bear nucleic acids, specifically short interfering RNA (siRNA) and plasmid DNA (pDNA). Since our goal is to design these particles for blood administration, their compatibility with blood is important in determining their therapeutic efficacy. We want to see if the nanoparticles will remain intact and if there will be interactions between the nucleic acid cargo and the cells in blood. We want to look at cytokine secretion from the blood cells as a measure of biocompatibility, and if changes in gene expression occurs as a result of exposure to the nanoparticles. There are no specific research subjects. We will obtain ‘pooled’ buffy coat or whole blood from CBS for our studies with no identification information. We plan to obtain further samples (mainly whole blood samples) from CBS in the coming year. With studies using commercial serum, we were able to show that the nanoparticles we designed were able to protect the drug cargo against degradative enzymes in the body and that nanoparticles remain functional in delivery of genes to the cells. The particles also remained stable but the aggregation studies were limited by the lack of red blood cells in the samples. A stable nanoparticle carrier for gene-based therapeutic agents will be developed upon successful completion of this project. We hope to show that our nanoparticles will not aggregate, will display controlled interactions with the cells of the blood component, elicit minimal cytokine response and minimal changes in gene expression, and deliver their cargo effectively to the cells. We plan to fine tune nanoparticle formulations so that nanoparticles that display the desired response with blood cells. Ultimately, we plan to develop nanoparticles that are tailored for specific applications, for example in leukemia therapy or therapy of other systemic diseases. 

Project ID: 2018.053b4r

Project Approval Date: 2018-12-13

City: Ottawa

Use of Animals: No

Canadian Blood Services Mandate: Does not benefit Canadian Blood Services

Summary: We hypothesize that a fibrin aptamer (DNA sequence) can be tagged to a fluorescent dye, CT contrast agent, or MRI contrast agent and can be developed to tag blood clots in vitro, laying the ground work for in vivo diagnosis of the location of bleeding such as in aneurysm rupture. This aptamer may be used to localize bleeding in clinical diseases such as stroke and ruptured intracranial aneurysms to facilitate early and appropriate treatment of these conditions. 

Project ID: 2019.001b4r

Project Approval Date: 2019-03-26 

City: Sudbury

Use of Animals: No

Canadian Blood Services Mandate: Does not benefit Canadian Blood Services

Summary: This study is able to investigate the toxicity of small molecules and their ability to act as anti-inflammatories on primary immune cells. These cells were extracted from the buffy coats of healthy, anonymous donors, which has been performed repeatedly for these studies. Once the primary peripheral mononuclear cells(PBMCs)have been extracted standard molecular biology techniques are then applied to study these cells under various treatment conditions. We have collected cells from four healthy donors (via CBS) which has been sufficient for experiments to date. We have successfully investigated different agents using PBMCs and our results demonstrate that these small molecules are non-toxic when used at doses that are able to induce both anti-cancer and anti-inflammatory effects. We are continuing with our preclinical investigations of these agents in various inflammatory conditions that represent either infection or autoimmune conditions to try and flush out the mechanisms of action to better understand these agents. The potential impact of the study it its continued development of new therapies for cancer, arthritis and potentially other inflammatory conditions.

Project ID: 2019.023b4r

Project Approval Date: 2019-07-26  

City: Vancouver

Use of Animals: No 

Canadian Blood Services Mandate: Does not benefit Canadian Blood Services

Summary: Our  group  is  developing  human  “organs  in  a  dish”.  Using  these  organs,  we  can  study  underlying  mechanism  of  diseases and we can test new therapies or drugs on these organ models which may help to replace or avoid animal testing at some point in the future. To achieve this, however, the organ models need a certain level of complexity and the implementation of immune cells is very important – not only to mimic certain diseases in the dish but also  to  determine  wanted  and  unwanted  effects  of  drugs.  Currently,  we  are  focusing  on  diseases  and  new  treatment  options  for  human  skin  and  human  lung.  To  generate  more  complex  organ  models,  we  plan  to  obtain/isolate certain immune cells from the donated blood, which we will then integrate into the organ models. This will then allow to study underlying mechanism of the respective diseases and may also allow to study drug effects in these models.

Project ID: 2019.028b4r

Project Approval Date: 2019-10-22 

City: Vancouver

Use of Animals: No

Canadian Blood Services Mandate: Transfusion medicine

Summary: Each year, >190,000 Canadians are diagnosed with cancer, and ~77,000 die from the disease. Genetic testing of archival tissue or freshly biopsied tumor tissue is required for most types of cancer to aid in diagnosis and treatment. While genetic testing is currently performed in tumor tissue, rapidly evolving technology will allow the majority of assays to be performed on circulating tumor DNA found in the blood. The advantages of a blood test (aka. Liquid biopsy) include reduced cost, reduced invasiveness and the ability to repeatedly test tumors as they evolve throughout the course of therapy. This project will develop a liquid biopsy assay to look for cancer mutations in the blood, as a diagnostic assay as well as a means to monitor patient response to treatment or cancer reoccurrence. We expect the assay will serve several provinces and research groups through our clinical testing laboratory, and help improve the management of cancer patients, and potentially reduce blood utilization.

Project ID: 2019.029b4r

Project Approval Date: 2019-12-11 

City: Vancouver

Use of Animals: No

Canadian Blood Services Mandate: Transfusion medicine

Summary: Red blood cells circulate in the bloodstream for approximately 120 days, delivering oxygen to all tissues in the body. A key property of red blood cells is their ability to squeeze into capillary blood vessels that are many times smaller than their own diameter. Donated red blood cells, collected by the Canadian Blood Service for use in blood transfusions, can be stored for up to 42 days. During this time, the deformability of these cells is degraded, impairing their ability to circulate in capillary blood vessels. As a result, the viability of these cells in transfusion recipients is reduced from 120 days to, in some cases, hours or minutes. Currently, there are no simple methods to measure the quality of donated blood units, as defined by how long they will circulate in transfusion recipients. Additionally, there appears to be significant differences between donors, where some donors could provide blood that circulate longer in recipients than other donors. Our research team have developed a method to analyze red blood cell deformability by sorting these cells based on their deformability in order to determine if the more deformable red blood cells circulate for longer in recipients than less deformable cells. Using an accelerated aging model of RBCs stored in plastic tubes, we found that donor RBCs had degradation profiles that were highly variable between donors, but consistent for each donor. Importantly, some donors showed significant loss of RBC deformability during storage, while other donors showed little or no storage induced loss of RBC deformability. We show that, in most cases, blood bags preserved RBC deformability during the 42-day storage window, while the degradation of RBC deformability in the subsequent 14 days were highly variable. This trend was mirrored in blood unit segments (attached to the same blood bag unit) with a strong correlation to the blood bag data. Strong correlations were also observed between blood bag and segment for MCV, MCHC and MCH, but not for hemolysis. Our results confirm that RBC deformability provides a potentially useful approach for donor-level screening to identify donors for whom the storage expiration window could potentially be lengthened.

Project ID: 2019.030b4r

Project Approval Date: 2019-10-09 

City: Toronto

Use of Animals: No 

Canadian Blood Services Mandate: Transplantation medicine

Summary: Macrophages and T cells are immune cells whose behavior can be altered by a wide range of biochemical cues presented by tissues and other cells. Both the presence and treatment of disease, in addition to transplantation procedures can alter macrophage and T cell behavior to worsen or improve patient recovery and prognosis. Traditionally, biomaterials including water-swollen hydrogels composed of natural sugars and proteins have been used as implants and vehicles for drug delivery. More recently, biomaterials themselves have been shown to have effects on immune cell recruitment and macrophage phenotype, raising the question of what is the ideal material to study these cells. Primary human macrophage and Tcell behaviour is not previously been studied in materials that represent the diseased extracellular matrix, which could reveal relevant behaviour that could support treatment of disease. We have developed a material called a cryogel, which is a hydrogel that is formed using a freezing technique to introduce pores. This new material is made from gelatin and hyaluronan, two components that are upregulated in a variety of disease microenvironments. The pore size and stiffness of the material can be modulated using chemistry that is used to covalently bind the components together. We used buffy coats from CBS donors to isolate monocytes which we then differentiate to macrophages and seed on our cryogels. We showed that the cryogel properties can influence cell migration, and we also show that the mechanism of migration through our gels is the same as that through in vivo tumour environments. We show that treatment with different proteins affects the migration capacity, which shows that we can use our cryogel to study relevant biological phenomenon. The first phase of this project has been published in Advanced Functional Materials. Our next stage of the project to be completed within one year will be co-culture with macrophages/T cells and diseased cells, to see the effects of cell-cell interactions on macrophage and T cell behaviour. This information could reveal drug targets, help us develop better disease models, and even discover insights to immune cell behaviour that could provide insights for transplantation research.

Project ID: 2019.036b4r

Project Approval Date: 2020-01-16 

City: Edmonton

Use of Animals: No

Canadian Blood Services Mandate: Transfusion medicine

Summary: Red cell concentrates (RCCs) can be cryopreserved and stored for up to ten years, allowing blood collection centers to maintain an inventory of units with rare red blood cell (RBC) phenotypes. International sharing of blood products allows blood centers to have increased access to rare RCC units. However, the unit must be thawed and deglycerolized prior to being shipped internationally. Once deglycerolized, the storage age of the RCC is 3 – 14 days. Logistically, this represents a challenge to ensure the unit arrives within this period for transfusion. Additionally, if upon arrival the unit is no longer required, it cannot be re-frozen and must be discarded. By shipping rare RCCs frozen, the loss of unused products can be prevented. To prepare RBCs for cryopreservation and to deglycerolize units prior to transfusion, collection centers around the world use the ACP-215 Automated Cell Processing System. The ACP-215 is relatively standardized but the deglycerolization protocol and bowl sizes can be modified which introduces variability in processing across collection centers. These variations, along with the collection method, manufacturing method, and additive/storage solutions used, increase the site-to-site disparity between products. An evaluation of these variances is necessary to compare the in vitro quality of blood products from different collection centers. In this study, collection centers will pool and split, glycerolize, and freeze ABO/Rh compatible units for shipment to all participating centers. Receiving centers will use their own standard ACP-215 protocol for processing to determine the compatibility of externally glycerolized products with local processing methods. The deglycerolized units will then undergo quality testing to ensure the RCCs meet local quality standards. Additionally, a quality comparison of locally cryopreserved, locally thawed products and externally cryopreserved, locally thawed products will indicate whether shipping frozen units will continue to provide high quality products that are safe for transfusion.

Project ID: 2020.003

Project Approval Date: 2020-03-20 

City: Ottawa

Use of Animals: No

Canadian Blood Services Mandate: Does not benefit Canadian Blood Services

Summary: In heart disease, LDL accumulate in the arteries, where they become modified, aggregated and retained. Such deposits of aggregated LDL (agLDL) can be recognized by macrophages, which initiate the inflammation in the arteries. This will lead to recruitment of blood cells, create lesions in the arteries, which eventually blocks bloodflow. We propose to research and understand the process, thereby exploring the options to prevent inflammation in the arteriesto decrease heart diseases. This study is entirely in vitro. We are using mouse macrophages to provide novel information on how atherosclerosis is initiated. There is no patient involvement. To study this project, we need to use LDL to make agLDL. LDL is isolated from human plasma, which we obtain from Canada Blood Service. The results will provide new insight for the development of plaque in the arteries, which leads to heart attacks. By understanding the process, medical interventions could be developed to lessen the effect of agLDL.

Project ID: 2020.005

Project Approval Date: 2020-02-26 

City: Edmonton

Use of Animals: No

Canadian Blood Services Mandate: Transfusion medicine

Summary: Red cell concentrates (RCCs) contain subpopulations of red blood cells (RBCs) at each stage of their120-day lifespan. The age distribution of these cells is influenced by donor characteristics; a better understanding of donor effects on RBC subpopulations can help improve the quality of stored red blood cell products. As RBCs age, they become denser from cell membrane loss and dehydration. Differences in RBC subpopulation age/density allows cells to be separated into young (less dense) and old (more dense) using density gradient separation methods.This study will select donors based on age (< 30 years or > 60 years) and sex (male or female). The pre-donation hemoglobin levels and donation frequency of the donors will also be collected for further characterization of the samples. Cells will be separated for quality analysis using a Percolldensity gradient separation technique. It is suspected that young, female donors who donate more frequently and have lower hemoglobin levels will have a larger subpopulation of young, less dense RBCs. Results of this study will improve our understanding of the effect of donor characteristics on blood product quality and may result in changes to how blood components are produced and transfused.

Project ID: 2020.010

Project Approval Date: 2020-02-20 

City: Quebec

Use of Animals: No

Canadian Blood Services Mandate: Transfusion medicine

Summary: Malaria is one of the world’s most common infectious diseases, with approximately 274 million cases each year and 470000 deaths, and thus represents one of the most devastating global public health problems. The lack of an effective vaccine, resistance to all commercial antimalarial drugs, combined with the small number of suitable new drugs against the malaria parasite demonstrate the urgent need for the development and implementation of novel intervention strategies in the form of drugs, vector control measures and an effective vaccine. Indeed, it is expected that if the trend in malaria prevalence stays on its current upwards course, the death rate could double in the next twenty years. Invasion of a red blood cell by Plasmodium falciparum parasites is an essential step in the malaria lifecycle and host response to parasite antigens are an important component of human malarial immunity. Consequently, the molecular players involved in erythrocyte invasion are key targets for both therapeutic and vaccine-based strategies to block parasite development. Several of these invasion proteins are stored in the apical complex of the parasite, a structure containing secretory organelles called dense granules, micronemes and rhoptries, and are released at different times during invasion. Because of its essential role, interfering with the generation of the apical complex represents a very attractive target for the design of a new kind of antimalarial. Our studies will focus on trying to understand how the parasite directs proteins to the different structures of the apical complex. Understanding this complex process will likely provide a wealth of new targets for the development of strategies to block apical complex generation and preventing malaria pathogenesis. This could also lead to the development of antibody-based rapid diagnostic tests for the detection of malaria 

Project ID: 2020.015

Project Approval Date: 2020-04-21 

City: Toronto

Use of Animals: No

Canadian Blood Services Mandate: Transfusion medicine

Summary: Cancer patients have been suffering various side effects from high-dose systemic administration of anti- cancer treatment. This project aims to address this issue by using the ability of platelets to home to cancer sites to deliver the drug directly to the tumor and avoid systemic toxicities. Platelets are the natural carrier of proteins used for cancer growth and replacing the cargo with a drug is an easy way to enrich the cancer environment with the therapeutic agent. This is different from nanoparticles which are not native particles and can accumulate in the body. We use a specific “hook” which attaches the drug to the inside of platelet α-granules. This internalization not only protects the drug from degradation, also reduces the drug systemic toxicity because no free drug is circulating through unintended organs. Finally, the on-site drug release, in addition to minimizing any off-target toxicities, allows the drug to circulate for a longer time, namely for the life span of the platelets (4-7 days). As designed the drug of interest needs to be loaded into donors’ platelets ex vivo. This is where platelet rich plasma purchased from Canadian Blood Services is employed. The technology has no requirement on the participants except to avoid non-steroidal anti-inflammatories such as aspirin or naproxen before blood donation. Continuing our research on the conjugated antibody in the previous year (2020), our work in 2021 showed that the drug “hook” can be attached onto small-molecules and that attaching the ‘hook’ does not affect the function of the drug. The conjugate can be successfully internalized into resting platelets with no platelet activation or aggregation, and the drug/’hook’ conjugates are actively sequestered in platelet α-granules. The loaded platelets were enriched on the tumor sites in tumor-bearing mice but not the spleen where platelets usually get recycled. In this coming year, we plan to establish the efficacy doses of the conjugate-loaded platelets. We continue to need support from Canadian Blood Services as a source of reliable platelet rich plasma. This drug delivery system based on loading platelets with a therapeutic agent is the first of its kind promising significant improvement in the therapeutic index of many present and future cancer therapies.

Project ID: 2020.016

Project Approval Date: 2020-03-31 

City: Toronto

Use of Animals: No

Canadian Blood Services Mandate: Transplantation medicine

Summary: The use of Canadian Blood products align with the Canadian blood services mandate in our full time educational programs in the past has relatively advanced the knowledge and practice in the areas of transplantation medicine . Also it is a valuable learning tool and assisted us to create high caliber professionals within the health care system. It will continually support us to train Professionals who will be efficient and minimize complications or delays which will benefit Health Care. Simulation based education is a proven instructional methodology that allows learners to make mistakes in authentic learning environment rather than learning on their real job where workload are high and lives are at risk. The Michener Institute of Education at UHN, Cardiovascular Perfusion (CVP) program is the only such program within the Province of Ontario. Providing students the ability to work with real human blood within the CVP bypass machine/Cell saver allowing student to gain tremendously valuable training. Cardiovascular perfusion is a profession primarily focused on supporting the normal cardio-pulmonary functions during open-heart surgery. The Medical Laboratory Science Program includes student training in the fields of Transfusion Medicine and Hematology. Students gain skills and experience within Transfusion Medicine through Laboratories requiring ABO typing, Cross and Match, Antibody investigations and Phenotyping. The Hematology labs allow students to learn how to handle the actual blood samples, also give opportunity to operate Complete Blood Cell Count (CBC) analyzers, perform and review blood cells smears and conduct tests utilizing Coagulation analyzers. Michener’s commitment to educating high quality health care practitioners and it is our number one priority. Using best practices in simulation-based education requires using “authentic” human blood.

Project ID: 2020.018

Project Approval Date: 2020-06-22 

City: Manitoba

Use of Animals: Yes

Canadian Blood Services Mandate: Transplantation medicine

Summary: The objective of this research project is to generate a “humanized” mouse model to study the early phases of HIV infection. Specifically, we will determine whether the biology of human white blood cells (e.g. T cells, B cells and monocytes) can be studied within a more physiological environment using a mouse model. The rationale is that the experimental conditions by which we study human white blood cells have an impact on their biology. Therefore, it is important to develop an animal model to study their behaviors within complex biological systems that closely recapitulates their natural setting. The goal is to understand how HIV infection alters the behavior and numbers of human white blood cells in blood, and to uncover new mechanisms that can be therapeutically targeted to suppress HIV replication. Our studies also have implications on understanding how HIV can impact transplantation of mice with human blood cells. To accomplish this, we will transplant 50 million human white cells from healthy blood donors (from buffy coats) and monitor their numbers in mice (e.g. blood, lymphoid organs). We will phenotypically characterize human white blood cells in blood for up to 4 weeks. After HIV infection of the animals, we will determine whether this mouse model can recapitulate many of the hallmarks of HIV infection in humans, such as profound T cell infection and death in the blood. We will also use this model to understand how HIV spreads from one part of the body to another. Taken together, we will develop a white blood cell transplantation mouse model to study the biology of human white blood cells, and how these behaviors are altered during HIV infection. In future studies, we will further develop these animal models to study other blood-borne pathogens.

Project ID: 2020.019

Project Approval Date: 2020-03-31 

City: Windsor

Use of Animals: 

Canadian Blood Services Mandate: Transplantation medicine

Summary: In this project we study the innate immune responses against cytomegalovirus (CMV) infection in healthy humans. Approximately, 40-70 of the population is latently infected with CMV. However, this virus can cause severe disease and complications in immunocompromised patients. The immune system of a healthy individual is very effective in preventing CMV disease. A type of white blood cell called the natural kill (NK) cells are particularly important in immunity against this virus. NK cells seek out and destroy virus infected cells. This is performed with the help of proteins called receptors found on the surface of the NK cells. Depending on the type of receptors present on the surface of NK cells, they can either provide immunity against CMV or be inhibited by the virus. CMV infection also causes changes in NK cell receptor composition to influence its activity. We seek to understand how receptors are modulated during CMV infection and which receptors are important for NK cell function to provide immunity against this virus. For this project we analyze NK cells isolated from blood of healthy donors. NK cell receptors and their activity will be compared between CMV-positive (latent infection) and CMV-negative (uninfected) individual. These studies will advance our understanding of how NK cells provide immunity against CMV and which receptors are important for this function of NK cells.

Project ID: 2020.025

Project Approval Date: 2020-07-03 

City: Edmonton

Use of Animals: No

Canadian Blood Services Mandate: Transfusion medicine

Summary: Cryopreserved red cell concentrates (RCCs) can be stored for transfusion for up to 30 years at temperatures below -65 °C.  During storage RCCs may experience unintentional warming to temperatures above -65 °C but below 0 °C.   During this type of warming  event ice crystals can grow larger leading to cell damage if not controlled. However, glycerol, the chemical agent used to freeze red blood cells (RBCs), can provide some protection against the  effects of ice recrystallization by inhibiting the growth ofice crystals in RCCs. Previous studies have demonstrated that multiple cycles of warming events with low concentrations of glycerol (15%) increase damage to RBCs where high concentrations of glycerol (40%) reduce this effect.

This study will evaluate the impact of unintentional warming  events on  RCCs by testing  the  quality of these products after the glycerol is removed.The first part of this study uses RCCs that have been pooled and split, cryopreserved, and exposed to various types of unintentional warming events that emulate real world scenarios that can happen during frozen storage. The second part of this study evaluates RCCs that have undergone a real world documented unintentional warming event. Results of this study will improve our understanding of what happens to the quality of RBCs that have been exposed to unintentional warming  events  and may result in changes to how blood components are stored and managed.

Project ID: 2020.031

Project Approval Date: 2020-06-23 

City: Richmond

Use of Animals: No

Canadian Blood Services Mandate: Does not benefit Canadian Blood Services

Summary: Pro-Lab is developing a blood test to determine if people have been exposed to Covid-19 by detecting antibodies that have developed to the virus in patient/donor blood. The donor samples will be used throughout the internal validatio  and  quality control studies on Latex agglutination, ELISA and Lateral flow assays to ensure assay performance optimization. 

Project ID: 2020.032

Project Approval Date: 2020-07-23 

City: Vancouver

Use of Animals: Yes

Canadian Blood Services Mandate: Transplantation medicine

Summary: Adoptive immunotherapy, such as chimeric antigen receptor (CAR) T cell therapy, is a new treatment approach that has been shown to induce deep remission in patients with a variety of blood cancers. These therapies are manufactured by isolating patient immune cells, genetically modifying them to recognize the tumour cells, expanding them to high numbers, then infusing them back into the patient to achieve deep cancer remissions. This study aims to 1) aid in the development of new immune cellular therapies and 2) determine the optimal timing and manufacturing processes used to produce highly effective and safe cell therapies. As these therapies are manufactured using human immune (blood) cells, donated blood products are essential for the development of these therapies. So far the study has focused on testing new CAR-T cell therapy products for multiple myeloma and the results will soon be presented at an upcoming international myeloma society (IMS) meeting. The study is also exploring ways of making more durable (persistent) CAR-T cell therapy products through more sophisticated manufacturing processes. It is expected that new, more effective, CAR-T cell therapies will be identified in this study and these new cell therapy products will improve patient outcomes across a variety of blood cancers.

Project ID: 2020.038

Project Approval Date: 2020-09-08 

City: Vancouver

Use of Animals: Yes

Canadian Blood Services Mandate: Transplantation medicine

Summary: Transplantation is used to treat end-stage organ failure, as well as cancers or other deficiencies of the immune system. In order for bodies to accept transplanted cells or organs, patients must take powerful immunosuppressive drugs for the rest of their lives. These drugs have many side effects and often do not effectively stop the immune responses that lead to transplant failure. Our lab is studying how a specialized type of white blood cell known as a T regulatory cell, or Treg, turns the immune system off. The normal function of Tregs is to prevent autoimmunity. We aim to learn how to harness the protective properties of Tregs so that they can be used as a therapy to prevent the undesired immune responses that cause transplant failure and/or autoimmunity. The goal is to find ways to track and manipulate Tregs therapeutically so that the need for transplants would be reduced (i.e. by decreasing autoimmunity which leads to end stage organ failure) as well as the need for life-long immunosuppressive drugs in patients who received a transplant. Overall the research will lead to the development of improved outcomes for patients who need, or who have had, a transplant.In the past year, discoveries which were made possible by the use of CBS blood products included:-Discovered that removal of a signaling protein known as PTEN can increase the effectiveness of Treg therapy -Found that assessing expression of a protein known as Helios is important for determining the purity of a Treg cell therapy product -Published our method on how to measure the ability of Tregs to suppress antigen presenting cells so that other investigators can easily use this assay in their studies.

Project ID: 2020.041

Project Approval Date: 2021-02-26 

City: Vancouver

Use of Animals: No

Canadian Blood Services Mandate: Transfusion medicine

Summary: Characterization of blood and blood components (platelets, red blood cells and plasma)manufactured by Canadian Blood Services is important for our understanding of their use for transfusion and for the development of new blood components. This study will perform studies that assess the quality and biology/functionality of blood and blood components routinely produced by Canadian Blood Services. Findings will improve our understanding of the components and serve as baseline data when exploring components in development.

This study will also assess blood components that are under development by Canadian Blood Services and not yet available for transfusion. For examples, leukoreduced whole blood, cold-stored platelets, platelets suspended in additive solutions, pathogen-reduced platelets, rejuvenated red blood cells, red blood cells suspended in alternative additive solution, and freeze-dried plasma, may be investigated during the course of this study. Finally, this study will explore the optimization of assays and/or new technology for studying the new components and establishing their quality in the context of transfusion practice.

Project ID: 2020.044

Project Approval Date: 2020-09-09 

City: Vancouver

Use of Animals: No

Canadian Blood Services Mandate: Transplantation medicine

Summary: The purpose of this study is to develop methods for cell separation based on microscopy images. Immunophenotyping is an expensive and error-prone process, despite significant efforts in standardizing staining, data collection and automation of analysis. Also, morphological differences between cells may not need to be apparent to the naked eye in order to be sufficient for cell classification. Consequently, we will develop an approach for using disaggregated cells, similar to the preparation employed in standard laboratory flow cytometry, to establish rapid and robust image-based cell phenotyping of cells. With this capability, it might be possible to develop a model for the 200+ known cell types in the human body in order to detect previously unknown phenotypes or detect phenotypic shifts that occur because of disease or treatment. Immune cells will be separated from whole blood and stained with a variety of fluorescent markers. We will assess the cell images and develop tools in order to predict cell phenotype and function. Finally, cells will be separated based on their phenotype and function from the microscopy images. We have developed a technology to separate cells based on microscopy using micropatterning of photo-reactive hydrogels to selectively retain unwanted cells in a microwell plate, which enable the extraction of target cells by simple pipetting. Our process is high-throughput and insensitive to cell type.

Project ID: 2020.047

Project Approval Date: 2020-10-01

City: Toronto

Use of Animals: Yes

Canadian Blood Services Mandate: Transplantation medicine

Summary: Graft-versus host disease (GVHD) following stem cell transplant is often a major limitation to the success of this therapy for treating blood cancers. Stem cell transplant can be curative, but 30-80% of patients develop acute GVHD within the first 100 days of transplant.1 While 30% of patients fail to respond to the current therapies, this study addresses discovering new therapeutic options for acute GVHD patients.

Project ID: 2020.051

Project Approval Date: 2020-11-04 

City: Vancouver

Use of Animals: No

Canadian Blood Services Mandate: Transplantation medicine

Summary: We investigate human embryonic and T cells in culture, for research purposes only and with no humans involved (besides the Canadian Blood Services source of the blood cells and the embryonic stem cell derivations that were done by other labs a long time ago). Our work is mainly on developing the best ways to culture these cells, to produce desired therapeutic cell types.

Project ID: 2020.052

Project Approval Date: 2020-12-15 

City: Vancouver

Use of Animals: Yes

Canadian Blood Services Mandate: Transfusion and Transplantation medicine

Summary: Hematopoietic Stem Cell Transplantation (HSCT), the infusion of a donors immune system into a recipient, is an important immune-based therapy for patients with high-risk and recurring blood cancers. However, HSCT is attributed to an increased risk of health complications, the most severe being chronic graft-versus-host-disease (cGvHD), in which foreign donor immune cells attack the recipient’s tissues. cGvHD occurs in 25% of pediatric and 60% of adult HSCT survivors. It is known to cause long-term, and often irreversible organ damage and has a 10-25% mortality rate. 

Our team has identified an increased number of a rare peripheral immune cell population, known as regulatory Natural Killer cells (NKreg), in transplant patients who failed to develop cGvHD. Further, other researchers have found that the transfer of expanded NK cells potentially decreases blood cancer relapse with no increase in GvHD. Thus, we hypothesize that the transfer of expanded NKreg cells after transplant represents a significant strategy to decrease cGvHD without increasing blood cancer relapse.To address our hypothesis, we have been pursuing three aims. First, we have been determining the function of NKreg cells associated with successful and failed suppression of cGvHD. Second, we have been optimizing the expansion of functional NKreg cells appropriate for human clinical trial. Third, we will evaluate the effect of expanded NKreg cells on cGvHD immune populations in both culture and mouse models. 

Project ID: 2020.057

Project Approval Date: 2021-01-13 

City: Ottawa

Use of Animals: Yes

Canadian Blood Services Mandate: Transplantation medicine

Summary: The goal of our research is to understand the role of MTF2 in blood cells. MTF2 is part of a protein complex that controls whether of some genes are turned on or off in blood cells. When MTF2 is not properly functioning in leukemia cells, we discovered that it is not able to properly control the expression certain genes. This results in genes being abnormally turned on, which in turn prevents the leukemia cells from responding to chemotherapy treatment. Using cord blood cells obtained from CBS, we demonstrated that using drugs to block a gene called MDM2, which is abnormally turned on, can help leukemia cells respond to chemotherapy. This idea is being tested in a clinical trial to help leukemia patients respond better to standard chemotherapy. We will continue to characterize the genes controlled by MTF2 to understand the function of the genes in blood cell development and leukemia. The results of these studies will advance our knowledge about the normal and abnormal development of blood cells that may be applied to developing new effective therapies to treat leukemia.

Project ID: 2020.058

Project Approval Date: 2020-12-17 

City: Prince George

Use of Animals: No

Canadian Blood Services Mandate: Transplantation medicine

Summary: This project supports the training of medical lab technologists in the field of transfusion science by providing experience for students in the Medical Laboratory Technology Science Program at the College of New Caledonia. Each student interacts with the blood and blood products from CBS during laboratory training and practice activities. These interactions help students to develop a foundation of skills, behaviours and attitudes that are essential for student progress to clinical training in hospital laboratories and transfusion departments. This experiential learning is essential to student development and is superior to the study of images and theoretical knowledge alone. The MLTS program orders packed red blood cells and occasionally frozen plasma to stock the CNC MLTS laboratory each semester.

Project ID: 2020.059

Project Approval Date: 2020-12-11 

City: Toronto

Use of Animals: Yes

Canadian Blood Services Mandate: Does not benefit Canadian Blood Services

Summary: Objective: To identify novel epigenetic modulators of human CD8+ T cell function.

Current progress: Through pooled lentiviral CRISPR screen in primary human CD8+ T Cells, we have identified novel regulators of T cell function. Our immediate steps are to validate these targets that positively regulate CD8+ T cell proliferation, activation and cytolytic function.

Significance: Results obtained in this study will identify new therapeutic avenues to enhance T cell function through epigenetic modulation. Given that studies are performed in human T cells, the results have the potential to be incorporated into adoptive transfer of engineered T cells into patients.In addition, to fostering scientific collaborations in Toronto, Ontario; scientific discoveries generated from this study have the potential in to be used as treatment for hematological and solid cancers by harnessing the immune system.

Project ID: 2021.003

Project Approval Date: 2020-12-11 

City: London

Use of Animals: No

Canadian Blood Services Mandate: Transplantation medicine

Summary: Transplantation improves the quality of life and survival of patients at a much lower cost than dialysis. However, only 40% of patients receive functional kidneys due to a lack of living organ donors. To increase the number of kidneys available for transplantation, surgeons are accepting organs from deceased donors. These kidneys are commonly stored in cold solution without oxygen. As a result, organs suffer injuries. Therefore, we need to develop a new method to preserve donated kidneys that address these issues. Our objective is to study the effects of blood on storing human donor kidneys for long periods of time (16 hours) at room temperature. We will also evaluate if certain drugs can maximize the preservation of these organs. Our main goal is to properly preserve kidneys coming from dead donors, to extend their lifetime after transplantation and to improve the quality of life of patients while reducing the cost of transplantation worldwide.

Project ID: 2021.006

Project Approval Date: 2021-05-13 

City: Edmonton

Use of Animals: No

Canadian Blood Services Mandate: Transfusion medicine

Summary: This study will examine the age-specific differences in red blood cell biology across multiple international blood services to inform how donor age and differences in blood component manufacturing may affect the quality of stored blood. Most studies examining blood product quality have not included samples from teenage blood donors yet these donors can contribute up 10 -25% of the blood supply. As teenage blood donors are vulnerable to iron deficiency which can affect the quality and storage of red blood cell products produced from their donations, characterizing age-specific differences in blood products must include this important blood donor group. This study will characterize what biological differences that exist between teenage (16 – 19 years) and adult (> 35 years) blood donors at four different international blood services.  

Project ID: 2021.007

Project Approval Date: 2021-03-09 

City: Vancouver

Use of Animals: No

Canadian Blood Services Mandate: Transfusion medicine

Summary: Platelets are an integral part of blood clotting, and used clinically to manage bleeding in patients suffering from a number of diseases. The short shelf life of platelets remains a major challenge in the management of platelet products. This project asks whether new tools for delivering non-native proteins and genetic material to platelets using state-of-the-art-lipid particles will lead to a new understanding of the processes that affect platelet activity during storage, and whether these tools can lead to modified platelet products. The long-term objective of this project is to improve the therapeutic capabilities and shelf life of existing platelet products by treatment with our lipid particles, reducing the demand on the current platelet product supply. This goal is directly in line with the Canadian Blood Services mandate to advance transfusion science.

Project ID: 2021.008

Project Approval Date: 2021-02-19 

City: Toronto

Use of Animals: No

Canadian Blood Services Mandate: No direct benefit to either transplantation or transfusion medicine

Summary: Osteoarthritis is a condition that affects millions of Canadians and that is a slow, gradual wearing down of tissue and bone that affects the movement and flexibility in your joints. As you get older, it is possible to develop osteoarthritis in your knees and other parts of your body. Presently, it can be difficult to determine how severe the condition is or how it will progress simply by evaluating symptoms and medical imaging alone. Currently, there is no highly effective clinical treatments or cures for this disease. The purpose of this study is to discover the effects of arthritis on the biology of cells, and the role of inflammation in osteoarthritis. Healthy samples are needed to be used as a ‘control’ or comparison with the arthritis samples in this study. We also want to find out if these blood cells can increase the effectiveness of cells found in the bone marrow (mesenchymal stromal cells) because these bone marrow cells can be used as a treatment for arthritis.

Project ID: 2021.009

Project Approval Date: 2021-02-18 

City: Toronto

Use of Animals: No

Canadian Blood Services Mandate: Transfusion medicine

Summary: This is a quality project. The blood product will be used to validate Control Rate Freezers (CRFs) for the purpose of implementing a new cryopreservation procedure for all stem cell products processed in the lab, in place of the current practice of dump freezing.

Project ID: 2021.013

Project Approval Date: 2021-03-24 

City: Edmonton

Use of Animals: No

Canadian Blood Services Mandate: Transfusion and Transplantation medicine

Summary: Red blood cells (RBCs) are possibly the most transfused blood component and the most widely stored cell type. While cold storage (+4 °C) of RBCs has been vastly improved in the last few decades with a standard storage time of 42 days in clinical settings, recent clinical retrospective as well as laboratory studies indicate that storage in excess of 14 days may alter the biochemical properties of RBCs and lead to inferior outcomes in patients. Here, we propose to develop an RBC preservation method based on our recent breakthrough in deep supercooling of aqueous solutions where we can achieve seemingly stable supercooling for large volumes stored at low temperatures (down to -20 °C) for long periods of time. Our eventual goal, by combining our deep supercooling approach with other advances in RBC storage, is to extend the acceptable storage period to a minimum of 100 days. This would relieve logistical constraints, reduce waste of donated blood, and alleviate concerns related to use of aged cold-stored RBCs.

Project ID: 2021.014

Project Approval Date: 2021-04-15 

City: Edmonton

Use of Animals: No

Canadian Blood Services Mandate: No direct benefit to either transplantation or transfusion medicine

Summary: There is an urgent unmet need for non-invasive, simple, but reliable tests to not only detect cancer at an early stage but to also estimate how aggressive the cancer may be. Such early prognostication would provide doctors important information to balance watch-and-wait paradigms with aggressive treatment strategies that may carry risks. Biopsies and pathology are the gold-standard but biopsies cannot be done too often, and there is considerable patient resistance to such procedures. As such, we aim to develop novel strategies for early prognostication of cancer from liquid biopsies. Blood tests have been widely researched but few blood biomarkers have early prognostic value. Circulating tumor cells (CTCs) have considerable prognostic value but are extremely sparse. We propose early prognostication using strategies to significantly amplify blood biomarker levels to enrich CTCs and other blood borne markers such as mRNAs for analysis. We would like to acquire blood samples from CBS to identify baseline expression levels of various blood borne biomarkers to assess their relative utility in our liquid biopsy assays.

Project ID: 2021.017

Project Approval Date: 2021-05-06 

City: Toronto

Use of Animals: No

Canadian Blood Services Mandate: No direct benefit to either transplantation or transfusion medicine

Summary: Neutrophilic dermatosis are typically painful, acute-on-chronic inflammatory skin diseases presenting with recurrent pustules and abscesses, leading to scarring and permanent disfigurement. The quality of life in the patients with such diseases, like Hidredinitis Supparitiva (HS) and Pyoderma Gangrenosum (PG) are severely impacted as result of the chronic pain, odorous discharge and disfigurement. The underlying etiology of these inflammatory diseases are presently unknown and current treatment is often inadequate or ineffective. It is critical to our understanding of pathogenesis of disease progression of neutrophilic dermatoses to identify biomarkers in both skin and blood, in order to develop targeted treatments, and ultimately a cure. Currently, there are drugs being developed to interfere with the hypothesised immunological mechanism underlying HS and PG. We will evaluate the effect of the newly generated drug family of IRAK4 degraders, as well as other therapeutic interventions, as a novel targeted therapeutic for the treatment for these diseases. The finding of this study will enable us to identify the biomarkers in HS disease and the efficacy of therapeutic intervention and ultimately find a cure. 

Project ID: 2021.019

Project Approval Date: 2021-07-07 

City: Vancouver

Use of Animals: No

Canadian Blood Services Mandate: No direct benefit to either transplantation or transfusion medicine

Summary: Early disease diagnosis is important for effective treatment, and for determining who can take part in trials for new treatments. This has been a major challenge in the field of neurological disease, notably for cases of dementia, brain injury, and spinal cord injury, where most clinical procedures involve using expensive imaging techniques. Emerging tools for early disease diagnosis include the use of “biomarkers”, which are disease-specific molecules in the body that can be detected in blood. Biomarkers are the focus of many research groups in developing simple and low-cost blood tests for disease diagnosis. However, there is a challenge in not only determining which biomarkers reflect these diseases best, but also determining how to measure them accurately so that they can be used in the future as diagnostic tools in the clinic. This project will prepare pooled blood samples from individual donors, collected by Canadian Blood Services, to create reference materials. These materials will be sent out to other laboratories to check specificity and reproducibility of new blood tests being developed for dementia, brain injury, and spinal cord injury, before they enter the clinic, and will become part of an international quality control program for neurological blood biomarkers.

Project ID: 2021.030

Project Approval Date: 2021-09-09 

City: Ralston

Use of Animals: No

Canadian Blood Services Mandate: No direct benefit to either transplantation or transfusion medicine

Summary: The impact of COVID-19 pandemic is horrifying and unprecedented. It is likely that the pandemic might not end any time soon due to the emergence of more and more SARS-CoV-2 variants all over the world. It is therefore imperative to rapidly develop, in parallel to vaccines, therapeutics against SARS-CoV-2 and its variants to fight the pandemic. Human monoclonal antibodies (mAbs) are safe and ideal therapeutics and most importantly, human mAbs can be rapidly developed to treat infectious diseases from the blood of convalescent or vaccinated donors. Current antibody therapies are usually administered intravenously due to the large amount of antibody required. The high dosage results in high cost and intravenous infusion, which requires hospital settings, making the mAb therapy hard be accessible by the developing countries. To solve this issue, it is desirable to develop highly potent human mAbs with lower dosages required, which will lead to cost reduction and being deliverable via intramuscular or subcutaneous injections. To this end, in this proposal, single memory B cells, which have undergone somatic mutations so as to secrete high-affinity SARS-CoV-2 spike protein-specific antibodies, in human blood samples from SARS- CoV-2 convalescent or vaccinated donors will be isolated through a series of separation steps, including memory B-cells enrichment, single memory B-cell sorting, selection of single memory B-cells secreting highly potent anti-SARS-CoV-2 antibodies by neutralization assay, PCR amplification of antibody genes from the selected single memory B cells. Lastly the amplified antibody genes will be expressed in mammalian expression system and then the recombinant human antibodies characterized.

Project ID: 2021.032

Project Approval Date: 2021-07-20 

City: Calgary

Use of Animals: No

Canadian Blood Services Mandate: No direct benefit to either transplantation or transfusion medicine

Summary: The blood provided will be used to grow the malaria parasite, Plasmodium falciparumand to develop diagnostic test for other species of the malaria parasite infecting human, including but not limited to P. vivax, P. ovale spp, P. malariae andP. Knowlesi. We study Plasmodium falciparum intra-erythrocytic stages. The parasite multiplies asexually in the host erythrocytesin vivo. In vitro growth of laboratory strains or cryopreserved clinical isolates relies on red blood cells in the appropriate culture media. Our laboratory is relying on Plasmodium falciparum culture for several projects. Canada Blood Services (CBS) blood will be use to grow the laboratory strains used as internal control, including but not limited to: 

• Genotyping study and parasite whole genome sequencing: developing genotyping and sequencing methods to elucidate parasite genotypes associated with clinical outcome to malaria disease and diagnostic performances.
• Treatment resistance of the malaria parasite: laboratory strains as internal control, and propagate clinical isolates for drug susceptibility testing.
• Point of care diagnostic test development: grow parasites at the desired parasitemia and prepare fresh contrived samples for diagnostic test development. These test will include sexual and asexual forms of the parasites.
• Parasite phenotype in relation to clinical outcome

Project ID: 2021.033

Project Approval Date: 2021-10-06 

City: Toronto

Use of Animals: No

Canadian Blood Services Mandate: Transplantation medicine

Summary: The liver is the only visceral organ in the body with a rapid regenerative capacity. Understanding the mechanisms that help regenerate the liver is critical to repairing the damaged liver in patients with liver failure. To better understand how normal liver regeneration occurs and how we can distinguish normal versus abnormal regeneration, we will examine in this study both liver and plasma samples undergoing regeneration. We have selected liver biopsy specimens identified as normal regeneration within the 3 first months following liver transplant, and we will request the retrieval of plasma samples that are matched or unmatched with these liver biopsies from the UHN Biobank. Liver biopsies have been used to characterize different cell types in the regeneration process and their molecular signatures, while the plasma will be used to extract exosomes, and identify particular circulating gene markers. We will process to the comparison of circulating markers of normal regeneration, defective regeneration (plasma from cirrhotic liver patients), and healthy controls (plasma of blood donors from Canadian Blood Services). In the prospective study, we will collect blood from both liver donors and liver transplant recipients to investigate the regeneration activity following hepatectomy and liver transplant.

Project ID: 2021.037

Project Approval Date: 2022-04-01 

City: Montreal

Use of Animals: No

Canadian Blood Services Mandate: Transfusion medicine

Summary: The blood stages of human Plasmodium falciparum parasites cause the pathology associated with malaria. Extracellular vesicles (EVs) are small, membrane-bound particles that have been implicated in the biomolecular mechanisms by which P. falciparum potentiates malaria pathology and may also use EVs to promote its survival within the human host. Naïve and P. falciparum-infected red blood cells (RBCs) release different EV subpopulations depending on the age of the RBCs and the intraerythrocytic life stage of the parasite. These subpopulations may further comprise different EV subtypes according to their size (small, medium, large) and/or biogenesis. Functional studies have shown that different EV subpopulations and subtypes target different host cells and effect specific functions within these target cells, which include immune cells, endothelial cells, and other erythrocytes. As with all EVs, P. falciparum-infected RBC-derived EVs mediate intercellular communication through their biomolecular cargo that includes nucleic acids, proteins, and lipids.We hypothesize that the target cell specificity of EV subpopulations and subtypes is determined by their biochemical cargo. This study will identify the biochemical distinctions and/or commonalities of EVs released from naïve, ring, trophozoite and schizont-infected RBCs, as well as EVs from different aged RBCs. These findings will provide insights into the biogenesis, target cell specificity and functions of the different EV subpopulations and subtypes.Human RBCs are essential in carrying out this study and the data will also provide insight into how age of the RBC will alter the cargo and biogenesis of EVs released from the cell’s surface.

Project ID: 2021.041

Project Approval Date: 2022-01-17 

City: Vancouver

Use of Animals: No

Canadian Blood Services Mandate: No direct benefit to either transplantation or transfusion medicine

Summary: The COVID-19 pandemic continues to be a public health emergency in Canada and around the world. The emergence of novel COVID-19 variants means that researchers must continue to develop novel assays for the detection and quantification of SARS-CoV-2, the virus that causes COVID-19. The overall goal of this project is to source convalescent plasma from donors infected with SARS-CoV-2 variants of concern (VOCs), variants of interest (VOIs), and other variant strains of importance. The convalescent plasma will be used to support a global program to construct standards and panels for serologic assays and especially assays designed to look at differentiation and quantitation of, as well as antibody response to, SARS-CoV-2 variants. This study is conducted in collaboration with public health officials at the BC Center for Disease Control, Vitalant Research Institute, and Duke Human Vaccine Institute.

Project ID: 2021.046

Project Approval Date: 2022-03-09 

City: Vancouver

Use of Animals: No

Canadian Blood Services Mandate: Transfusion medicine

Summary: Platelets are the blood cells which are responsible for forming blood clots. Platelet transfusion is a lifesaving treatment for bleeding, such as on the battlefield or on automobile accidents, and is an important therapy for certain blood disorders and chemotherapy. Platelets have short shelf-lives because their current storage bags tend to activate them, leading to rapid clearance and poor transfusion outcomes. Furthermore, the stored cells can be easily contaminated by bacteria, which leads to severe sickness and platelet wastage. This project aims to develop platelet storage bags which extend the shelf-life of these cells and prevents bacterial colonization. We will screen a list of different coating materials to devise a formulation which most optimally extends platelet shelf life while being hostile to bacteria. We will then implement this coating into our bags and assess the short- and long-term effects of storage on the cells. Successfully extending the viability and protecting the quality of platelets will improve the outcome of their transfusion, directly saving the lives of recipients. This project may also save millions of tax payer dollars in the long run by limiting wastage of platelets due to expiration and contamination.

Project ID: 2021.047

Project Approval Date: 2022-02-03 

City: Halifax

Use of Animals: Yes

Canadian Blood Services Mandate: Transplantation medicine

Summary: Our research focuses on a type of white blood cell, called natural killer (NK) cells. NK cells are poorly understood because they are complicated, and the genes that control their function are variable between people. Still, it is known that NK cells can directly detect and kill cancer cells, or infected cells, while also directing other immune cells to help. For these reasons, we think it is important to better understand both the genetics and functions of NK cells. NK cells are already being targeted or used in clinical trials. They can be expanded virtually limitlessly and adoptively transferred between people without causing damage to healthy tissues, or they can be guided by antibodies and other therapies that control their function. Still, studies have not yet considered how the different features of NK cells factor into their activity against each person’s disease. Likely as a result, the outcomes of clinical trials have been variable. Our research goals are to understand how the genetics and phenotypes of NK cells predict their functions for direct killing and polarizing immune responses. Understood, this will inform how NK cells can be directed to control cancer and other diseases. In particular, we are interested in two gene families that have the most control over NK cell function: the Killer Immunoglobulin-like receptors (KIR) and their ligands, groups of human leukocyte antigens (HLA). NK cells use these KIR proteins to engage with conserved ligands that are found on groups of HLA molecules, so we measure only these ligands (not specific HLA alleles). In patients undergoing bone marrow transplantation or hematopoietic stem cell transplantation, where HLA alleles are matched between donors and recipients, it is already known that the donor’s KIR genes (which are on a separate chromosome from HLA and need not be matched) predict for cancer relapse or control. We think that NK cells’ KIR genes likewise have impacts on NK cell function in adoptive transfer of mature cells. Hence, our work is important to Canadian Blood Services in that it may inform immunotherapies, including stem cell transplantation and in the future, adoptive cell therapies.

Project ID: 2022.002

Project Approval Date: 2022-01-28 

City: Petawawa

Use of Animals: No

Canadian Blood Services Mandate: Transfusion medicine

Summary: The majority of potentially salvageable combat casualties die from hemorrhage. Whole blood is recommended for the resuscitation of bleeding combat casualties by the Canadian Forces Health Services (CFHS) endorsed by the Damage Control Resuscitation (DCR) clinical practice guideline, and is being used for the resuscitation of civilian trauma patients in the United States. Canadian Blood Services (CBS)is in the process of submitting a licensing application to Health Canada for a whole blood component that, if approved, could be used by the CFHS. Health Canada is expected to approve this new blood component in fiscal year 2022/23 and Canadian Blood Services anticipates making this component available shortly after to health care organizations, including CFHS operations. This study is being conducted to evaluate the quality characteristics of this new blood component during an exercise simulating the CFHS unique operational and logistical environment. Findings from this study will support the use of whole blood components manufactured at CBS by the Canadian Armed Forces.

Project ID: 2022.003

Project Approval Date: 2022-02-25 

City: Vancouver

Use of Animals: No

Canadian Blood Services Mandate: No direct benefit to either transplantation or transfusion medicine

Summary: If someone acquires HIV, they will require lifelong treatment with antiretroviral medications. Some of the older HIV medications are good at controlling the virus, but have side effects that hurt the cells in our body. Some newer HIV medications have been linked to weight gain in adults, or developmental problems in babies. We want to study the new HIV medications, and their effect on the cells in our own body. We want to see if there is a difference in how our immune system responds and if there is damage to our cells when they are exposed to the HIV medication. It is important to know, as HIV and HIV medication are both lifelong. To do this research, we require a large quantity of blood cells to test to follow the effects of five different medications daily over 12 days. This is only possible with the help of Canadian Blood Services. We will take the blood, take out the immune cells, and treat them with HIV medication for a period of time. Afterwards, we will look for any damage the cells have sustained, and any delay in their immune response.

Project ID: 2022.009

Project Approval Date: 2022-05-04 

City: Winnipeg

Use of Animals: No

Canadian Blood Services Mandate: No direct benefit to either transplantation or transfusion medicine

Summary: The request for packed red blood cells and whole blood is not being made with the intent of conducting research, but rather, to use the material in the preparation of an artificial whole blood matrix resembling that of the newborn infant. This will facilitate the preparation of control blood specimens to be used in validating the analytical results of all testing procedures used to screen newborn infants for inherited metabolic diseases, endocrine dysfunction, and SCIDS (Severe Combined Immunodeficiency Syndrome). The Manitoba Newborn Screening program has been in operation for over 50 years improving the lives of newborn infants and continues to be one of the best programs in the country. We request the help of your organization and that of your generous donors, to allow our program to continue to provide high quality service for the newborns of Manitoba by supplying red blood cells and whole blood. Since the current approval for study REB 2017.004 is expiring April 3, 2022, this application is being submitted to allow for continuation of our current Newborn Screening program quality assurance and improvement practices of making the blood control specimens.

Project ID: 2022.012

Project Approval Date: 2022-05-13 

City: Waterloo

Use of Animals: No

Canadian Blood Services Mandate: No direct benefit to either transplantation or transfusion medicine

Summary: This study is to validate the clearance performance of an experimental hemodiafiltration set-up used in Ecuador, the location of the client, proposed to repurpose existing hemodialysis machines to perform hemodiafiltration in order to improve clearance performance. This study aims to validate these claims or to make recommendations based on the results in the case where the performance proves to be lower than expected. If proven successful, the client, Quantum Medical, may move forward with commercialization to bring a low-cost solution to treat chronic kidney disease in Ecuador. 

Project ID: 2022.014

Project Approval Date: 2022-05-18 

City: Calgary

Use of Animals: No

Canadian Blood Services Mandate: Transfusion medicine

Summary: 

What question are you trying to answer?

Exposure to high oxygen levels is harmful to the preterm infant and contributes to retinopathy of prematurity (ROP), a retinal disease that can result in blindness. ROP affects approximately 120 babies per year in Calgary. Several studies have found a marked increase in the occurrence of ROP following blood transfusions though the cause is not clear. Adult packed red blood cells (pRBC) in transfused blood bind oxygen less tightly than fetal red blood cells present in the preterm infant. Therefore, a newborn maintained at the same oxygen saturation, which reflects bound oxygen, would be expected to have more unbound oxygen (PaO2) following a pRBC transfusion. In clinical practice, we use continuous oxygen saturation (SaO2) as a surrogate marker of oxygenation. However, the PaO2 is the more physiologically relevant variable as it reflects the oxygen that is freely available to the cells.

How will the answer to your question help your research field?  

Quantifying the shift in oxygen binding following red blood cell transfusions is an important first step in eventually customizing SaO2 targets for each infant based on their blood’s oxygen binding properties and transfusion needs. We will use information from this study to design a future randomized control trial examining the effects of adjusting SaO2 targets following blood transfusions on reducing the incidence and severity of ROP.  

Project ID: 2020.016 

Project Approval Date: 2023-04-03 

City: Vancouver 

Use of Animals: No 

Canadian Blood Services Mandate: Transfusion Medicine 

Summary: In Canada, there is no upper age limit for blood donors, and people with diabetes (high blood sugar) are eligible to donate blood. The aims of this study are: 

1. To evaluate how well blood sugar is controlled in donors with diabetes and estimate how many donors who have not self-reported diabetes have high blood sugar.  
2. To evaluate the quality of parts of the blood donation in donors of different ages, and in donors with diabetes, during the time period when blood is normally stored before transfusion. Some studies suggest that there are differences in blood cells and plasma proteins in people with diabetes. Blood from older donors also differs from younger donors. Here, we will characterize blood products across the age range of eligible donors, and from individuals with different types of diabetes to determine how these products differ, and whether their storage performance is impacted. Better understanding how donor characteristics affect blood products will ensure the safe storage and use of these products and may also contribute to our understanding of aging and diabetes in the Canadian population. 

Project ID: 2022.020

Project Approval Date: 2022-07-11 

City: Vancouver

Use of Animals: No

Canadian Blood Services Mandate: No direct benefit to either transplantation or transfusion medicine

Summary: With the successful development of messenger ribonucleic acid (mRNA) vaccines against COVID-19, the field of RNA therapeutics has evolved. Due to the instability of RNA, these molecules require a vehicle for delivery to the cells in the human body and the current leading candidates for such deliveries include polymeric vehicles, lipid-nanoparticles, and nano emulsions. In the context of vaccine development, there is evidence that these vehicles provoke an immune response but the exact mechanisms are unknown. This project will use human peripheral blood mononuclear cells (PBMC) from whole blood to explore the human immune responses to these delivery systems. We hypothesize that using human PBMCs as a model system to study innate immune cellular pathways involved in recognizing and processing the vehicle-RNA-loading complexes would lead to improved formulations of the current RNA-based vaccines, improved clinical translatability of RNA delivery vehicles and eventually the development of RNA delivery vehicles based on therapeutic needs. The specific objective of this project is to gain a preliminary understanding of cellular pathways involved in recognizing and processing vehicle-RNA-loading complexes in immune cells. Once we have gained some understanding of the relevant pathways, we will begin altering our formulations to allow more effective delivery of therapeutic RNAs.

Project ID: 2022.021

Project Approval Date: 2022-07-214

City: Edmonton 

Use of Animals: No 

Canadian Blood Services Mandate: No direct benefit to either transplantation or transfusion medicine 

Summary: Molecular self-assembly ofpeptides is proving to be a powerful technique for forming novel materials that allow the incorporation of biofunctional motifs into complex nanostructures that are formed using benign assembly processes. In particular, the ion-complementary self-assembling peptide RADARADARADARADA (RADA4, type 1: -+;II:--++) has been shown to self-assemble into nanofibers that subsequently develop into a viscoelastic 3D matrix in aqueous solution. Recent work indicates that RADA4 may provide a robust nanomedicine platform, with traits ideal for soft -tissue applications: peptide sequence programmability, non-cytotoxic, elicit a minimal inflammatory response (mechanism unknown), applicable to non-invasive therapies, in situ gelation, 3-D cellular activities, localized drug delivery, and an internal hydration of up to 99.5% w/v water. Plasma protein interactions with foreign surfaces is the fate determining step that dictates the application of implant biomaterials, no previous work could be found that discussed the interaction between peptide self-assembled materials and plasma proteins. Moreover, there are very few studies that have attempted to correlate the physicochemical properties of peptide based materials to protein adsorption (denaturation, activation) that is so critical to a myriad of deleterious host responses (thrombosis, inflammation, etc.).Therefore, this research program will focus upon elucidating the fundamental effects of the amino acid content of self-assembling peptides on plasma protein interactions (adsorption, denaturation, activation) as well as platelet responses. It is thought that this information is of fundamental importance for understanding the molecular level mechanisms responsible for initiation of deleterious host responses to foreign materials, and may provide the 'design criteria' required for further advances in biomaterial development.  

Project ID: 2022.025 

Project Approval Date: 2022-07-26 

City: Halifax 

Use of Animals: No 

Canadian Blood Services Mandate: No direct benefit to either transplantation or transfusion medicine 

Summary: Balancing protein production and degradation is crucial for cellular health. Disruption of such balance is connected to many diseases, including autoimmune, neurodegenerative diseases, cardiac dysfunction, cancer, and viral infections. Proteasome is one of the main protein complexes driving protein degradation in cells. Therefore, alteration of proteasome function has great therapeutic potential for treating multiple diseases. The development of such approaches requires a thorough understanding of how the proteasome functions in cells. Our lab discovered that a protein called RACK1 binds to the proteasome and alters its activity. The goal of this research is to determine how RACK1 controls proteasome function via their interaction. Red blood cells have been used as a proteasome source in several studies. Therefore, proteasomes will be purified from human red blood cells obtained via Canadian Blood Services. RACK1 will be expressed and purified from bacteria. The purified proteasome and RACK1 will be subjected to cryogenic-electron-microscopy to determine where the two proteins interact. This information is essential for determining how proteasome function changes with or without RACK1 binding in cells. The outcome of this study is to provide new mechanistic insight to help guide the future development of therapies that alter proteasome activity to treat different diseases. 

Project ID: 2022.028 

Project Approval Date: 2022-09-09 

City: Toronto 

Use of Animals: No 

Canadian Blood Services Mandate: Transplantation Medicine 

Summary: Immune-based adoptive cellular therapies are a promising treatment modality for cancer patients. In its current form, blood-derived T cells are isolated, genetically modified with cancer targeting receptors such as T cell receptors (TCRs) or chimeric antigen receptors (CARs) and reinfused into patients for treatment. Development of these therapies have relied on the use of human blood from healthy donors who provide the immune cells for genetic modification and testing. The Canadian Blood Services through their Blood4Research program can be a reliable source of human blood for further development of these therapies. Though adoptive cellular therapy against blood cancers has been established, the development of successful therapies targeting solid tumors remains a major obstacle, particularly because highly specific and safe anti-cancer receptors have yet to be found. Recently, we have developed innovative approaches to identify cancer targeting TCRs and CARs. Using these approaches, our proposal seeks to test the ability of these receptors to kill cancer cells using blood derived T cells from healthy donor immune cells. It is hoped that upon completion of this proposal, we will have identified candidate anti-cancer receptors suitable for safe and effective adoptive cellular therapies against cancer. 

Project ID: 2022.031 

Project Approval Date: 2022-08-25 

City: Waterloo 

Use of Animals: No 

Canadian Blood Services Mandate: No direct benefit to either transplantation or transfusion medicine 

Summary: Community-led contaminant biomonitoring projects were conducted in the communities of the Dehcho and Sahtu regions in the Northwest Territories, and Old Crow, Yukon from 2016-2020. These projects involved the collection of human blood, analyzed for contaminants, including aluminum. Exposure to high aluminum levels can cause health effects including bone disease, anemia, and affect the lungs, and nervous system. Average blood aluminum levels in Old Crow blood samples were more than three times higher than reference ranges for the general Canadian population, and some samples were higher than levels associated with health effects. During the investigation into these levels, sources of possible sample contamination by aluminum were identified, including aluminum-bearing collection needles and caps on sample containers. This project was designed to reproduce the blood sampling process several times with a single sourced blood sample to measure the variability in aluminum levels between samples. This process will help us understand whether, and how frequently, blood can become contaminated by aluminum during the sampling process. The results of this study will help community members understand whether the blood aluminum levels measured as part of this project reflect actual levels in the community, and will inform future studies into aluminum exposure in the area. 

Project ID: 2022.032

Project Approval Date: 2022-11-28 

City: Ottawa

Use of Animals: No

Canadian Blood Services Mandate: No direct benefit to either transplantation or transfusion medicine

Summary: Heart disease remains one of the biggest causes of death and hospitalization in Canada. The build up of plaque in arteries is called atherosclerosis, which occurs over time and is closely linked to risk factors such as obesity, high cholesterol, high blood pressure and smoking. Our research focuses on the cells that form the arterial plaque and how changes in cellular metabolism can cause and potentially help limit the size and severity of the plaque. To study this, we expose cells in a petri dish to the types of factors that they would be exposed to in the body, mainly cholesterol. To do this, we isolate cholesterol from human donor blood and use it to mimic the conditions that lead to atherosclerosis and heart disease. This ability lets us test the importance and potential therapeutic value of different metabolic and immune pathways in cells.  

Project ID: 2022.034 

Project Approval Date: 2022-11-07 

City: Oshawa 

Use of Animals: No 

Canadian Blood Services Mandate: Transfusion Medicine 

Summary: Red blood cells (RBCs) are the most numerous cells in our body. However, we now know that they have a specialized cellular machinery that regulates a wide range of their functions. During this period, they are constantly exposed to a variety of environmental challenges such as oxidative stress, which affect their physiological homeostasis. When RBCs fail to counter these stressors, they are injured and are captured by specialized phagocytic cells in the spleen and liver. This feature becomes more prominent in RBCs stored in the blood bank wherein they are increasingly primed to undergo cell death.The molecular mechanisms underpinning these observations remain unclear. Using a wide array of cellular and molecular biology tools as well as systems biology approaches, our research will examine different signaling networks in RBCs.We will test this by studying their patterns of survival during storage under blood bank as well as in vitro conditions. The basic science knowledge gained from this work will improve our understanding of how RBCs thrive and adapt under different environmental challenges. The results from these studies may have future applications in improving RBC product quality in blood banking. 

Project ID: 2022.038 

Project Approval Date: 2022-11-22 

City: Richmond 

Use of Animals: No 

Canadian Blood Services Mandate: No direct benefit to either transplantation or transfusion medicine 

Summary: Neutrophils-the most abundant leukocytes in the circulation system- are the key players in the modulation of the immune response. The role of neutrophils in the tumour microenvironment (TME) is multifactorial and largely depends on the context. The tumour and other resident immune cells can affect neutrophil polarization towards a pro (N2)- or anti (N1)-tumorigenic role. Thus, the characterization of the neutrophil population within TME and the discovery of the compounds that can engage neutrophils in tumour killing will be beneficial for the broad spectrum of cancer patients. This study aims to obtain a characterization profile of the neutrophil population in the context of various in vitro tumour models. Acquired data can help us develop new therapeutic targets that will provide engagement of neutrophils in anti-tumour response.​  

Project ID: 2022.039 

Project Approval Date: 2023-01-10 

City: Hamilton 

Use of Animals: No 

Canadian Blood Services Mandate: Transplantation Medicine 

Summary: ​​In this project we utilize red blood cells to deliver drugs more efficiently and more sustainably to patients. We have already demonstrated the high potential of functionalized red blood cells in several pilot studies. Fighting bacterial contamination and or immunization against Covid-19 were successfully demonstrated and we have developed a suite of advanced methods to manipulate the red blood cells membrane on the nanoscale. We can sensitize red blood cells to specific targets in the human body and produce hybrid cells that can carry different pharmaceutical loads. In the next step, we will expand the application portfolio by incorporating nutrients, anti-cancer drugs and neurological drugs in three projects. Each project goes along with extensive characterization and testing of the synthesized functionalized red blood cells. The donated blood helps to: 1) Optimize our methods to ensure a rapid and personalized synthesis of functionalized red blood cells. 2) characterize of functionalized erythrocytes to verify size, shape, structure and affinity. 3) verify the compatibility of different blood groups. 4) test the synthesized red blood cells using cell-lines. The study's outcomes will benefit the Canadian Blood Services by investigating novel cellular therapies. 

Project ID: 2022.040 

Project Approval Date: 2023-02-03 

City: Vancouver 

Use of Animals: No 

Canadian Blood Services Mandate: Transfusion Medicine 

Summary: The purpose of the current research proposal is to use chemistry and biomedical engineering tools to develop novel chemical products and methods to address current clinical issues in blood transfusion. To address these needs, we hypothesize that the prevention of immunological rejection of donor blood is of critical importance in transfusion medicine, as unintentional mismatching of red blood cells (RBC) remains one of the most common causes of serious and sometimes fatal adverse reactions. In addition, shortage of blood supply in rare blood transfusion remains a major challenge to our blood bank system as well as for platelets. Also, we are working on developing new chemical molecules which could prevent rejection of transplanted organs so that patients can survive longer as well as on developing novel polymers coatings with antibacterial and antithrombotic properties for blood bags and catheters. To perform this research, we need fresh human blood and its components so that we can test our new drug molecules and methods before being tested in animals and in humans directly. 

Project ID: 2022.046 

Project Approval Date: 2023-02-03 

City: Vancouver 

Use of Animals: Yes 

Canadian Blood Services Mandate: Transfusion Medicine 

Summary: Virus-mediated outcomes involve an intricate sequence of events that ultimately culminate in the alteration of host cell processes. Cellular activation is a well-known contributor to disease progression which may be explained in the context of direct virus-cell interactions or virusdependent manipulation of host cell factors. The aim of this study is to therefore answer questions surrounding what happens when a virus encounters circulating blood cells and proteins. This study will help generate knowledge in three key areas. The anticipated results may: (1) help in the understanding of how viruses infect cells; (2) provide insight and ideas into how to maintain blood product safety; and (3) lead to new ways of predicting or treating virus induced pathology. 

Project ID: 2022.048 

Project Approval Date: 2023-02-03 

City: Vancouver 

Use of Animals: Yes 

Canadian Blood Services Mandate: Transfusion Medicine 

Summary: The aim of this study is to answer questions surrounding three unique areas of interest. The first project focuses on identifying new roles and uses for well-known molecules involved in blood clot formation that also help to dissolve the clot after it is no longer needed.  The second project is working toward understanding how common viruses can use certain clotting molecules to help infect cells. The third project endeavors to learn about normal blood clotting by identifying the specific differences that exist in patients with known blood clotting abnormalities. These studies may lead to new ways of predicting, diagnosing or treating heart disease, help in the understanding of how viruses infect cells, and identify novel genetic alterations that underline coagulation disorders. 

Project ID: 2022.049 

Project Approval Date: 2023-02-21 

City: Toronto 

Use of Animals: No 

Canadian Blood Services Mandate: Transplantation Medicine 

Summary: Liver transplantation is life-saving for patients with end-stage liver disease or advanced liver cancer. Unfortunately, there are not enough organ donors to meet the demand.Currently, strict criteria are applied to select donor livers. In donors with risk factors for poor liver graft function, the liver is often rejected for transplantation. Being able to assess liver function before the organ is transplanted might allow us to safely use livers which are currently rejected for transplantation. Ex vivo (meaning “outside the body”) assessment of donor livers is a new method to estimate function before exposing patients to excessive risks. In an ex vivo circuit, warm, oxygenated blood and nutrients are circulated through the donor liver for several hours. This is known as ex vivo perfusion. This allows the organ function to be monitored but also may offer the possibility of repairing injured grafts. 

This basic research study aims to develop a model of ex vivo human liver perfusion, to compare ex vivo liver perfusion with standard cold preservation, and to explore repairing poor or injured livers during ex vivo perfusion. Organs included in this study will have been turned down for transplantation according to current standard criteria and made available for research. None of the organs utilized in this study will be transplanted.