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SINGLE-CELL TRANSCRIPTOMICS OF HUMAN PANCREATIC ISLETS IN DIABETES AND ΒETA CELL DIFFERENTIATION

Weng, Chen
http://orcid.org/0000-0003-0801-7037
http://rave.ohiolink.edu/etdc/view?acc_num=case1612882103714773

Abstract Details

2021, Doctor of Philosophy, Case Western Reserve University, Genetics.
The pancreatic islet contains multiple endocrine (hormone-producing) cells that play critical roles in glucose homeostasis. Failure of the islet cells, especially the insulin-secreting β cells, is central to the development of diabetes. However, the molecular landscapes of the human pancreatic islet in normal and disease states remain unclear. Here we performed large-scale single-cell RNA-Seq for both adult human islets and stem-cell-derived β-like cells in differentiation. We first analyzed the transcriptome of 39,905 single islet cells from 9 human donors and observed distinct β cell heterogeneity trajectories associated with obesity or type II diabetes (T2D). We therefore developed RePACT, a sensitive single-cell analysis algorithm to identify both common and specific signature genes for obesity and T2D. We mapped both β cell-specific genes and disease signature genes to the insulin regulatory network identified from a genome-wide CRISPR screen. Our integrative analysis discovered the previously unrecognized roles of the cohesin loading complex and the NuA4/Tip60 histone acetyltransferase complex in regulating insulin transcription and release. Second, we generated 95,308 single-cell transcriptomes for the entire differentiation process from hESCs to β-like cells and reconstructed a lineage tree to study temporally 10 regulated genes. We identified so-called `switch genes’ at the branch points of endocrine/non-endocrine cell fate choices, revealing insights into the mechanisms of differentiation-promoting reagents, such as NOTCH and ROCKII inhibitors, thus providing the framework for improved differentiation protocols. Over 20% of all detectable genes are activated multiple times during differentiation, even though their enhancer activation is usually unimodal, indicating extensive gene reuse driven by different enhancers. We also identified a stage-specific enhancer in the TCF7L2 diabetes GWAS locus that drives a transient wave of gene expression in pancreatic progenitors. Taken together, this thesis work provides comprehensive single cell data resources and insights into islet biology and diabetes.
Fulai Jin (Advisor)
Li Yan (Advisor)
Ann Harris (Committee Chair)
Peter Scacheri (Committee Member)
Jonathan Haines (Committee Member)
162 p.
Genetics
Single-cell; Transcriptomics; Diabetes; Stem cell; Beta-cell;

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Citations

  • Weng, C. (2021). SINGLE-CELL TRANSCRIPTOMICS OF HUMAN PANCREATIC ISLETS IN DIABETES AND ΒETA CELL DIFFERENTIATION [Doctoral dissertation, Case Western Reserve University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=case1612882103714773

    APA Style (7th edition)

  • Weng, Chen. SINGLE-CELL TRANSCRIPTOMICS OF HUMAN PANCREATIC ISLETS IN DIABETES AND ΒETA CELL DIFFERENTIATION. 2021. Case Western Reserve University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=case1612882103714773.

    MLA Style (8th edition)

  • Weng, Chen. "SINGLE-CELL TRANSCRIPTOMICS OF HUMAN PANCREATIC ISLETS IN DIABETES AND ΒETA CELL DIFFERENTIATION." Doctoral dissertation, Case Western Reserve University, 2021. http://rave.ohiolink.edu/etdc/view?acc_num=case1612882103714773

    Chicago Manual of Style (17th edition)

case1612882103714773
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This open access ETD is published by Case Western Reserve University School of Graduate Studies and OhioLINK.