Novel Observations From Next-Generation RNA Sequencing of Highly Purified Human Adult and Fetal Islet Cell Subsets

被引:232
作者
Blodgett, David M. [1 ]
Nowosielska, Anetta [2 ]
Afik, Shaked [3 ]
Pechhold, Susanne [1 ]
Cura, Anthony J. [1 ]
Kennedy, Norman J. [4 ]
Kim, Soyoung [2 ]
Kucukural, Alper [5 ]
Davis, Roger J. [4 ,6 ]
Kent, Sally C. [1 ]
Greiner, Dale L. [2 ]
Garber, Manuel G. [3 ]
Harlan, David M. [1 ]
diIorio, Philip [2 ]
机构
[1] Univ Massachusetts, Sch Med, Dept Med, Diabet Ctr Excellence, Worcester, MA 01605 USA
[2] Univ Massachusetts, Sch Med, Program Mol Med, Diabet Ctr Excellence, Worcester, MA USA
[3] Univ Massachusetts, Sch Med, Program Mol Med, Program Bioinformat, Worcester, MA USA
[4] Univ Massachusetts, Sch Med, Program Mol Med, Worcester, MA USA
[5] Univ Massachusetts, Sch Med, Dept Biochem & Mol Pharmacol, Worcester, MA USA
[6] Univ Massachusetts, Sch Med, Howard Hughes Med Inst, Worcester, MA 01605 USA
基金
美国国家卫生研究院;
关键词
GENOME-WIDE ASSOCIATION; GENE-EXPRESSION; HIGH-THROUGHPUT; ARCHITECTURE; ALPHA; REPRESSION; MORTALITY; PANCREAS; PACKAGE; BURDEN;
D O I
10.2337/db15-0039
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
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.
引用
收藏
页码:3172 / 3181
页数:10
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