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 DM, Nowosielska A, Afik S, Pechhold S, Cura AJ, Kennedy NJ, Kim S, Kucukural A, Davis RJ, Kent SC, Greiner DL, Garber M, Harlan DM, diLorio PJ. (2015). Novel Observations From Next-Generation RNA Sequencing of Highly Purified Human Adult and Fetal Islet Cell Subsets. Program in Bioinformatics and Integrative Biology Publications. https://doi.org/10.2337/db15-0039. Retrieved from https://escholarship.umassmed.edu/bioinformatics_pubs/76