Journal
STEM CELLS AND DEVELOPMENT
Volume 27, Issue 13, Pages 898-909Publisher
MARY ANN LIEBERT, INC
DOI: 10.1089/scd.2017.0160
Keywords
small molecule screening; immature beta cells; murine embryonic stem cells; glucose sensors; postnatal young islets
Funding
- National Institutes of Health [R01DK081587, R01DK099734, P30CA33572]
- Oxnard Foundation
- Ella Fitzgerald Foundation
- Wanek Family Project of Type 1
- California Institute for Regenerative Medicine (CIRM)
- Department of Defense (DoD) through National Defense Science & Engineering Graduate Fellowship (NDSEG) Program
- National Science Foundation [DMR 1506483]
- Division Of Materials Research
- Direct For Mathematical & Physical Scien [1506483] Funding Source: National Science Foundation
Ask authors/readers for more resources
Pluripotent stem cells may serve as an alternative source of beta-like cells for replacement therapy of type 1 diabetes; however, the beta-like cells generated in many differentiation protocols are immature. The maturation of endogenous beta cells involves an increase in insulin expression starting in late gestation and a gradual acquisition of the abilities to sense glucose and secrete insulin by week 2 after birth in mice; however, what molecules regulate these maturation processes are incompletely known. In this study, we aim to identify small molecules that affect immature beta cells. A cell-based assay, using pancreatic beta-like cells derived from murine embryonic stem (ES) cells harboring a transgene containing an insulin 1-promoter driven enhanced green fluorescent protein reporter, was used to screen a compound library (NIH Clinical Collection-003). Cortisone, a glucocorticoid, was among five positive hit compounds. Quantitative reverse transcription-polymerase chain reaction analysis revealed that glucocorticoids enhance the gene expression of not only insulin 1 but also glucose transporter-2 (Glut2; Slc2a2) and glucokinase (Gck), two molecules important for glucose sensing. Mifepristone, a pharmacological inhibitor of glucocorticoid receptor (GR) signaling, reduced the effects of glucocorticoids on Glut2 and Gck expression. The effects of glucocorticoids on ES-derived cells were further validated in immature primary islets. Isolated islets from 1-week-old mice had an increased Glut2 and Gck expression in response to a 4-day treatment of exogenous hydrocortisone in vitro. Gene deletion of GR in beta cells using rat insulin 2 promoter-driven Cre crossed with GR(flox/flox) mice resulted in a reduced gene expression of Glut2, but not Gck, and an abrogation of insulin secretion when islets were incubated in 0.5mM d-glucose and stimulated by 17mM d-glucose in vitro. These results demonstrate that glucocorticoids positively regulate glucose sensors in immature murine beta-like cells.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available