Novel Observations From Next-Generation RNA Sequencing of Highly Purified Human Adult and Fetal Islet Cell Subsets
Department of Medicine; Diabetes Center of Excellence; Program in Molecular Medicine; Department of Biochemistry and Molecular Pharmacology; Program in Bioinformatics and Integrative Biology
Biochemistry | Bioinformatics | Cell Biology | Computational Biology | Endocrinology, Diabetes, and Metabolism | Integrative Biology | Systems Biology
Understanding distinct gene expression patterns of normal adult and developing fetal human pancreatic alpha- and beta-cells is crucial for developing stem cell therapies, islet regeneration strategies, and therapies designed to increase beta-cell function in patients with diabetes (type 1 or 2). Toward that end, we have developed methods to highly purify alpha-, beta-, and delta-cells from human fetal and adult pancreata by intracellular staining for the cell-specific hormone content, sorting the subpopulations by flow cytometry, and, using next-generation RNA sequencing, we report the detailed transcriptomes of fetal and adult alpha- and beta-cells. We observed that human islet composition was not influenced by age, sex, or BMI, and transcripts for inflammatory gene products were noted in fetal beta-cells. In addition, within highly purified adult glucagon-expressing alpha-cells, we observed surprisingly high insulin mRNA expression, but not insulin protein expression. This transcriptome analysis from highly purified islet alpha- and beta-cell subsets from fetal and adult pancreata offers clear implications for strategies that seek to increase insulin expression in type 1 and type 2 diabetes. long as the work is properly cited, the use is educational and not for profit, and the work is not altered.
DOI of Published Version
Diabetes. 2015 Sep;64(9):3172-81. doi: 10.2337/db15-0039. Epub 2015 Apr 30. Link to article on publisher's site
Blodgett, David M.; Nowosielska, Anetta; Afik, Shaked; Pechhold, Susanne; Cura, Anthony J.; Kennedy, Norman J.; Kim, Soyoung; Kucukural, Alper; Davis, Roger J.; Kent, Sally C.; Greiner, Dale L.; Garber, Manuel; Harlan, David M.; and diLorio, Philip J. Jr., "Novel Observations From Next-Generation RNA Sequencing of Highly Purified Human Adult and Fetal Islet Cell Subsets" (2015). Program in Bioinformatics and Integrative Biology Publications and Presentations. 76.