{"id":{"repo_id":"penn","oai_identifier":"oai:repository.upenn.edu:20.500.14332/28440"},"canonical_url":"https://search.dev.ndltd.org/etd/penn/oai:repository.upenn.edu:20.500.14332/28440","repository":{"repo_id":"penn","name":"University of Pennsylvania","base_url":"https://repository.upenn.edu/server/oai/request"},"display":{"title":"Targeted Epigenetic Editing to Increase Adult Pancreatic Β-Cell Proliferation","abstract":"β-cell replacement therapy is potentially a curative approach in treating diabetes, as demonstrated by the success of pancreatic islet transplantation in type 1 diabetes. However, there are an insufficient number of organ donors to meet the demand of this disease, which is increasing in prevalence. One strategy to increase the supply of human β-cells for transplantation in type 1 diabetics, or to increase residual β-cell mass in type 2 diabetics, is to induce human β-cell replication. This strategy has not been implemented clinically because adult human β-cells are largely quiescent and the capacity for proliferation decreases with age. I hypothesized that changes in DNA methylation contribute to the age-related decline in proliferative capacity in human β-cells, and that altering the DNA methylome in a targeted manner could improve proliferative capacity. To investigate this hypothesis, I sought to profile the β-cell across the human lifespan, and to develop tools that permit targeted DNA methylation modifications and efficiency in measuring DNA methylation. I conducted RNA-Seq and whole-genome bisulfite sequencing (WGBS) to profile the aging human β-cell transcriptome and DNA methylome. I found that there are significant changes in gene expression with age, and in DNA methylation, particularly at islet-specific active enhancers. Further, I developed transcription activator-like effector (TALE) fusion proteins conjugated to DNA methyltransferases (DNMTs) and demonstrated that targeting TALE-DNMTs to the promoter of the CDKN2A locus, encoding the cell cycle inhibitor p16, increases proliferation in primary human fibroblasts. Finally, I developed BisPCR2, a novel technique for preparing targeted bisulfite next-generation sequencing libraries, which greatly improves the efficiency in which DNA methylation can be measured at target regions. I demonstrated the utility of this tool to validate genome-wide findings of type 2 diabetes CpG risk loci. Together, these novel datasets and epigenetic tools poise the β-cell regeneration field to investigate targeted epigenetic modifications as a strategy to improve proliferative capacity of adult human β-cells.","abstract_html":"β-cell replacement therapy is potentially a curative approach in treating diabetes, as demonstrated by the success of pancreatic islet transplantation in type 1 diabetes. However, there are an insufficient number of organ donors to meet the demand of this disease, which is increasing in prevalence. One strategy to increase the supply of human β-cells for transplantation in type 1 diabetics, or to increase residual β-cell mass in type 2 diabetics, is to induce human β-cell replication. This strategy has not been implemented clinically because adult human β-cells are largely quiescent and the capacity for proliferation decreases with age. I hypothesized that changes in DNA methylation contribute to the age-related decline in proliferative capacity in human β-cells, and that altering the DNA methylome in a targeted manner could improve proliferative capacity. To investigate this hypothesis, I sought to profile the β-cell across the human lifespan, and to develop tools that permit targeted DNA methylation modifications and efficiency in measuring DNA methylation. I conducted RNA-Seq and whole-genome bisulfite sequencing (WGBS) to profile the aging human β-cell transcriptome and DNA methylome. I found that there are significant changes in gene expression with age, and in DNA methylation, particularly at islet-specific active enhancers. Further, I developed transcription activator-like effector (TALE) fusion proteins conjugated to DNA methyltransferases (DNMTs) and demonstrated that targeting TALE-DNMTs to the promoter of the CDKN2A locus, encoding the cell cycle inhibitor p16, increases proliferation in primary human fibroblasts. Finally, I developed BisPCR2, a novel technique for preparing targeted bisulfite next-generation sequencing libraries, which greatly improves the efficiency in which DNA methylation can be measured at target regions. I demonstrated the utility of this tool to validate genome-wide findings of type 2 diabetes CpG risk loci. Together, these novel datasets and epigenetic tools poise the β-cell regeneration field to investigate targeted epigenetic modifications as a strategy to improve proliferative capacity of adult human β-cells.","abstract_has_math":false,"creators":["Bernstein, Diana Lynne"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Klaus H. Kaestner"],"committee_chairs":[],"committee_members":[],"year":2015,"date_issued":"2015-01-01","date_published":"2015-01-01","updated_at":"2026-07-24T03:45:38Z","subjects":[],"languages":["en"],"rights":["Diana Lynne Bernstein"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://repository.upenn.edu/handle/20.500.14332/28440","outbound_label":"Repository record","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Klaus H. 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However, there are an insufficient number of organ donors to meet the demand of this disease, which is increasing in prevalence. One strategy to increase the supply of human β-cells for transplantation in type 1 diabetics, or to increase residual β-cell mass in type 2 diabetics, is to induce human β-cell replication. This strategy has not been implemented clinically because adult human β-cells are largely quiescent and the capacity for proliferation decreases with age. I hypothesized that changes in DNA methylation contribute to the age-related decline in proliferative capacity in human β-cells, and that altering the DNA methylome in a targeted manner could improve proliferative capacity. To investigate this hypothesis, I sought to profile the β-cell across the human lifespan, and to develop tools that permit targeted DNA methylation modifications and efficiency in measuring DNA methylation. I conducted RNA-Seq and whole-genome bisulfite sequencing (WGBS) to profile the aging human β-cell transcriptome and DNA methylome. I found that there are significant changes in gene expression with age, and in DNA methylation, particularly at islet-specific active enhancers. Further, I developed transcription activator-like effector (TALE) fusion proteins conjugated to DNA methyltransferases (DNMTs) and demonstrated that targeting TALE-DNMTs to the promoter of the CDKN2A locus, encoding the cell cycle inhibitor p16, increases proliferation in primary human fibroblasts. Finally, I developed BisPCR2, a novel technique for preparing targeted bisulfite next-generation sequencing libraries, which greatly improves the efficiency in which DNA methylation can be measured at target regions. I demonstrated the utility of this tool to validate genome-wide findings of type 2 diabetes CpG risk loci. Together, these novel datasets and epigenetic tools poise the β-cell regeneration field to investigate targeted epigenetic modifications as a strategy to improve proliferative capacity of adult human β-cells."]},{"key":"dc:description.degree","label":"Dc Description Degree","values":["Doctor of Philosophy (PhD)"]},{"key":"dc:format.mimetype","label":"Dc Format Mimetype","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Targeted Epigenetic Editing to Increase Adult Pancreatic Β-Cell Proliferation"]}]}],"canonical_facts":{"dc:contributor.advisor":["Klaus H. Kaestner"],"dc:creator":["Bernstein, Diana Lynne"],"dc:date":["2023-05-17T15:52:11.000"],"dc:date.accessioned":["2023-05-22T16:41:06Z"],"dc:date.available":["2016-02-09T00:00:00Z"],"dc:date.issued":["2015-01-01"],"dc:description.abstract":["β-cell replacement therapy is potentially a curative approach in treating diabetes, as demonstrated by the success of pancreatic islet transplantation in type 1 diabetes. However, there are an insufficient number of organ donors to meet the demand of this disease, which is increasing in prevalence. One strategy to increase the supply of human β-cells for transplantation in type 1 diabetics, or to increase residual β-cell mass in type 2 diabetics, is to induce human β-cell replication. This strategy has not been implemented clinically because adult human β-cells are largely quiescent and the capacity for proliferation decreases with age. I hypothesized that changes in DNA methylation contribute to the age-related decline in proliferative capacity in human β-cells, and that altering the DNA methylome in a targeted manner could improve proliferative capacity. To investigate this hypothesis, I sought to profile the β-cell across the human lifespan, and to develop tools that permit targeted DNA methylation modifications and efficiency in measuring DNA methylation. I conducted RNA-Seq and whole-genome bisulfite sequencing (WGBS) to profile the aging human β-cell transcriptome and DNA methylome. I found that there are significant changes in gene expression with age, and in DNA methylation, particularly at islet-specific active enhancers. Further, I developed transcription activator-like effector (TALE) fusion proteins conjugated to DNA methyltransferases (DNMTs) and demonstrated that targeting TALE-DNMTs to the promoter of the CDKN2A locus, encoding the cell cycle inhibitor p16, increases proliferation in primary human fibroblasts. Finally, I developed BisPCR2, a novel technique for preparing targeted bisulfite next-generation sequencing libraries, which greatly improves the efficiency in which DNA methylation can be measured at target regions. I demonstrated the utility of this tool to validate genome-wide findings of type 2 diabetes CpG risk loci. Together, these novel datasets and epigenetic tools poise the β-cell regeneration field to investigate targeted epigenetic modifications as a strategy to improve proliferative capacity of adult human β-cells."],"dc:description.degree":["Doctor of Philosophy (PhD)"],"dc:format.mimetype":["application/pdf"],"dc:identifier.uri":["https://repository.upenn.edu/handle/20.500.14332/28440"],"dc:language":["en"],"dc:rights":["Diana Lynne Bernstein"],"dc:title":["Targeted Epigenetic Editing to Increase Adult Pancreatic Β-Cell Proliferation"],"dc:type":["Dissertation/Thesis"]},"updated_at":"2026-07-24T03:45:38Z"}