Novel Observations From Next-Generation RNA Sequencing of Highly Purified Human Adult and Fetal Islet Cell Subsets.
Understanding distinct gene expression patterns of normal adult and developing fetal human pancreatic α- and β-cells is crucial for developing stem cell therapies, islet regeneration strategies, and therapies designed to increase β-cell function in patients with diabetes (type 1 or 2). Toward that end, we have developed methods to highly purify α-, β-, and δ-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 α- and β-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 β-cells. In addition, within highly purified adult glucagon-expressing α-cells, we observed surprisingly high insulin mRNA expression, but not insulin protein expression. This transcriptome analysis from highly purified islet α- and β-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.
Department of Medicine, Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, MA.
David M
DM
Blodgett
Program in Molecular Medicine, Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, MA.
Anetta
A
Nowosielska
Program in Molecular Medicine, Program in Bioinformatics, University of Massachusetts Medical School, Worcester, MA.
Shaked
S
Afik
Department of Medicine, Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, MA.
Susanne
S
Pechhold
Department of Medicine, Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, MA.
Anthony J
AJ
Cura
Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA.
Norman J
NJ
Kennedy
Program in Molecular Medicine, Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, MA.
Soyoung
S
Kim
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA.
Alper
A
Kucukural
Program in Molecular Medicine, University of Massachusetts Medical School, and Howard Hughes Medical Institute, Worcester, MA.
Roger J
RJ
Davis
Department of Medicine, Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, MA.
Sally C
SC
Kent
Program in Molecular Medicine, Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, MA.
Dale L
DL
Greiner
Program in Molecular Medicine, Program in Bioinformatics, University of Massachusetts Medical School, Worcester, MA.
Manuel G
MG
Garber
Department of Medicine, Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, MA david.harlan@umassmemorial.org.
David M
DM
Harlan
Program in Molecular Medicine, Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, MA.
Philip
P
diIorio
D000293:Adolescent; D000328:Adult; D002675:Child, Preschool; D005260:Female; D005333:Fetus / Q000166:cytology*; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D050416:Glucagon-Secreting Cells / Q000378:metabolism*; D006801:Humans; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D007515:Islets of Langerhans / Q000166:cytology / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D011247:Pregnancy; D011262:Pregnancy Trimester, Second; D012313:RNA / Q000235:genetics*; D017423:Sequence Analysis, RNA; D019864:Somatostatin-Secreting Cells / Q000378:metabolism*; D055815:Young Adult
D012313:RNA