{"id":{"repo_id":"toronto-retro","oai_identifier":"oai:utoronto.scholaris.ca:1807/149618"},"canonical_url":"https://search.dev.ndltd.org/etd/toronto-retro/oai:utoronto.scholaris.ca:1807/149618","repository":{"repo_id":"toronto-retro","name":"University of Toronto","base_url":"https://utoronto.scholaris.ca/server/oai/request"},"display":{"title":"Characterizing the Immunogenicity of Islet-like Cells derived from HLA-edited Human Pluripotent Stem Cells","abstract":"Although there is much hope for the therapeutic use of islet-like cells generated from human pluripotent stem cells, important challenges limit the clinical translation of this technology. In this thesis, I describe my efforts to address two of these challenges: the inability to control the composition of the islet-like cells generated, and the susceptibility of stem cell-derived islets to allorejection. I first describe methods to improve differentiation into cells bearing the successive phenotypes of primitive streak, definitive endoderm, and pancreatic progenitors. I next explore mechanisms that regulate the endocrine composition of human pluripotent stem cell-derived islets, finding that events both prior to and concurrent with endocrine commitment have important lineage-modulating effects. With these findings, I develop strategies to generate islet-like cells enriched for islet-like (ISL1+) or enterochromaffin-like (ISL1-SLC18A1+) cells. Finally, I leverage these new protocols to study how HLA proteins affect the immunogenicity of human pluripotent stem cell-derived islets. Using human pluripotent stem cells edited to lack cell-surface expression of class I and class II HLA complexes, which retain the ability to generate islet-like cells with insulin-secreting functionality, I find that CD8+ and CD4+ T cell allorecognition requires the presence of cell-surface HLA. Importantly, islet-like cells generated from HLA-edited human pluripotent stem cells demonstrate the potential to evade antibody-mediated rejection in vitro and T cell-mediated rejection in vitro and in vivo. These findings could be used to inform the development of new methods to generate islet-like cells more efficiently and safely for the treatment of type I diabetes.","abstract_html":"Although there is much hope for the therapeutic use of islet-like cells generated from human pluripotent stem cells, important challenges limit the clinical translation of this technology. In this thesis, I describe my efforts to address two of these challenges: the inability to control the composition of the islet-like cells generated, and the susceptibility of stem cell-derived islets to allorejection. I first describe methods to improve differentiation into cells bearing the successive phenotypes of primitive streak, definitive endoderm, and pancreatic progenitors. I next explore mechanisms that regulate the endocrine composition of human pluripotent stem cell-derived islets, finding that events both prior to and concurrent with endocrine commitment have important lineage-modulating effects. With these findings, I develop strategies to generate islet-like cells enriched for islet-like (ISL1+) or enterochromaffin-like (ISL1-SLC18A1+) cells. Finally, I leverage these new protocols to study how HLA proteins affect the immunogenicity of human pluripotent stem cell-derived islets. Using human pluripotent stem cells edited to lack cell-surface expression of class I and class II HLA complexes, which retain the ability to generate islet-like cells with insulin-secreting functionality, I find that CD8+ and CD4+ T cell allorecognition requires the presence of cell-surface HLA. Importantly, islet-like cells generated from HLA-edited human pluripotent stem cells demonstrate the potential to evade antibody-mediated rejection in vitro and T cell-mediated rejection in vitro and in vivo. These findings could be used to inform the development of new methods to generate islet-like cells more efficiently and safely for the treatment of type I diabetes.","abstract_has_math":false,"creators":["Misra, Paraish Shivam"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":"Physiology","school":null,"contributors":[],"advisors":["Nostro, M. Cristina","Yuen, Darren A"],"committee_chairs":[],"committee_members":[],"year":2024,"date_issued":"2024-06","date_published":"2024-06","updated_at":"2026-07-27T21:28:18Z","subjects":["Allorejection","Diabetes","Insulin","Islets","Pluripotent stem cells","Transplantation"],"languages":[],"rights":["Attribution-NonCommercial 4.0 International"],"rights_urls":["http://creativecommons.org/licenses/by-nc/4.0/"],"identifier_entries":[]},"links":{"outbound_url":"https://hdl.handle.net/1807/149618","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Nostro, M. 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In this thesis, I describe my efforts to address two of these challenges: the inability to control the composition of the islet-like cells generated, and the susceptibility of stem cell-derived islets to allorejection. I first describe methods to improve differentiation into cells bearing the successive phenotypes of primitive streak, definitive endoderm, and pancreatic progenitors. I next explore mechanisms that regulate the endocrine composition of human pluripotent stem cell-derived islets, finding that events both prior to and concurrent with endocrine commitment have important lineage-modulating effects. With these findings, I develop strategies to generate islet-like cells enriched for islet-like (ISL1+) or enterochromaffin-like (ISL1-SLC18A1+) cells. Finally, I leverage these new protocols to study how HLA proteins affect the immunogenicity of human pluripotent stem cell-derived islets. Using human pluripotent stem cells edited to lack cell-surface expression of class I and class II HLA complexes, which retain the ability to generate islet-like cells with insulin-secreting functionality, I find that CD8+ and CD4+ T cell allorecognition requires the presence of cell-surface HLA. Importantly, islet-like cells generated from HLA-edited human pluripotent stem cells demonstrate the potential to evade antibody-mediated rejection in vitro and T cell-mediated rejection in vitro and in vivo. These findings could be used to inform the development of new methods to generate islet-like cells more efficiently and safely for the treatment of type I diabetes."]},{"key":"dc:description.degree","label":"Dc Description Degree","values":["Ph.D."]},{"key":"dc:title","label":"Title","values":["Characterizing the Immunogenicity of Islet-like Cells derived from HLA-edited Human Pluripotent Stem Cells"]}]}],"canonical_facts":{"dc:contributor.advisor":["Nostro, M. Cristina","Yuen, Darren A"],"dc:contributor.department":["Physiology"],"dc:creator":["Misra, Paraish Shivam"],"dc:date":["2024-06"],"dc:date.accessioned":["2025-11-07T05:12:52Z"],"dc:date.issued":["2024-06"],"dc:description.abstract":["Although there is much hope for the therapeutic use of islet-like cells generated from human pluripotent stem cells, important challenges limit the clinical translation of this technology. In this thesis, I describe my efforts to address two of these challenges: the inability to control the composition of the islet-like cells generated, and the susceptibility of stem cell-derived islets to allorejection. I first describe methods to improve differentiation into cells bearing the successive phenotypes of primitive streak, definitive endoderm, and pancreatic progenitors. I next explore mechanisms that regulate the endocrine composition of human pluripotent stem cell-derived islets, finding that events both prior to and concurrent with endocrine commitment have important lineage-modulating effects. With these findings, I develop strategies to generate islet-like cells enriched for islet-like (ISL1+) or enterochromaffin-like (ISL1-SLC18A1+) cells. Finally, I leverage these new protocols to study how HLA proteins affect the immunogenicity of human pluripotent stem cell-derived islets. Using human pluripotent stem cells edited to lack cell-surface expression of class I and class II HLA complexes, which retain the ability to generate islet-like cells with insulin-secreting functionality, I find that CD8+ and CD4+ T cell allorecognition requires the presence of cell-surface HLA. Importantly, islet-like cells generated from HLA-edited human pluripotent stem cells demonstrate the potential to evade antibody-mediated rejection in vitro and T cell-mediated rejection in vitro and in vivo. These findings could be used to inform the development of new methods to generate islet-like cells more efficiently and safely for the treatment of type I diabetes."],"dc:description.degree":["Ph.D."],"dc:identifier.uri":["https://hdl.handle.net/1807/149618"],"dc:rights":["Attribution-NonCommercial 4.0 International"],"dc:rights.uri":["http://creativecommons.org/licenses/by-nc/4.0/"],"dc:subject":["Allorejection","Diabetes","Insulin","Islets","Pluripotent stem cells","Transplantation"],"dc:title":["Characterizing the Immunogenicity of Islet-like Cells derived from HLA-edited Human Pluripotent Stem Cells"],"dc:type":["Thesis"]},"updated_at":"2026-07-27T21:28:18Z"}