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Anti-diabetic action of all-trans retinoic acid and the orphan G protein coupled receptor GPRC5C in pancreatic β-cells

Amisten, Stefan LU ; Al-Amily, Israa Mohammad; Soni, Arvind LU ; Hawkes, Ross; Atanes, Patricio; Persaud, Shanta Jean; Rorsman, Patrik LU and Salehi, S Albert LU (2017) In Endocrine Journal 64(3). p.325-338
Abstract

Pancreatic islets express high levels of the orphan G-protein coupled receptor C5C (GPRC5C), the function of which remains to be established. Here we have examined the role of GPRC5C in the regulation of insulin secretion and β-cell survival and proliferation using human and mouse pancreatic islets. The expression of GPRC5C was analysed by RNA-sequencing, qPCR, western blotting and confocal microscopy. Insulin secretion and cell viability were determined by RIA and MTS assays, respectively. GPRC5C mRNA expression and protein level were reduced in the islets from type-2 diabetic donors. RNA sequencing in human islets revealed GPRC5C expression correlated with the expression of genes controlling apoptosis, cell survival and proliferation.... (More)

Pancreatic islets express high levels of the orphan G-protein coupled receptor C5C (GPRC5C), the function of which remains to be established. Here we have examined the role of GPRC5C in the regulation of insulin secretion and β-cell survival and proliferation using human and mouse pancreatic islets. The expression of GPRC5C was analysed by RNA-sequencing, qPCR, western blotting and confocal microscopy. Insulin secretion and cell viability were determined by RIA and MTS assays, respectively. GPRC5C mRNA expression and protein level were reduced in the islets from type-2 diabetic donors. RNA sequencing in human islets revealed GPRC5C expression correlated with the expression of genes controlling apoptosis, cell survival and proliferation. A reduction in Gprc5c mRNA and protein expression was observed in islets isolated from old mice (>46 weeks of age) compared to that in islets from newborn (<3 weeks) mice. Down-regulation of Gprc5c led to both moderately reduced glucose-stimulated insulin release and also reduced cAMP content in mouse islets. Potentiation of glucose-stimulated insulin secretion concomitant with enhanced islet cAMP level by all-trans retinoic acid (ATRA) was attenuated upon Gprc5c-KD. ATRA also increased [Ca+2 ]i in Huh7-cells. Gprc5c over expression in Huh7 cells was associated with increased ERK1/2 activity. Gprc5c-KD in clonal MIN6c4 cells reduced cell proliferation and in murine islets increased apoptosis and the sensitivity of primary islet cells to a cocktail of pro-apoptotic cytokines. Our results demonstrate that agents activating GPRC5C represent a novel modality for the treatment and/or prevention of diabetes by restoring and/or maintaining functional β-cell mass.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Diabetes, Insulin release, Orphan GPCR, RAIG2, RAIG3
in
Endocrine Journal
volume
64
issue
3
pages
14 pages
publisher
Japan Endocrine Society
external identifiers
  • scopus:85016566049
ISSN
0918-8959
DOI
10.1507/endocrj.EJ16-0338
language
English
LU publication?
yes
id
cad17e33-b9ef-42ad-8354-3601355beb3e
date added to LUP
2017-04-19 10:53:23
date last changed
2018-01-07 11:59:36
@article{cad17e33-b9ef-42ad-8354-3601355beb3e,
  abstract     = {<p>Pancreatic islets express high levels of the orphan G-protein coupled receptor C5C (GPRC5C), the function of which remains to be established. Here we have examined the role of GPRC5C in the regulation of insulin secretion and β-cell survival and proliferation using human and mouse pancreatic islets. The expression of GPRC5C was analysed by RNA-sequencing, qPCR, western blotting and confocal microscopy. Insulin secretion and cell viability were determined by RIA and MTS assays, respectively. GPRC5C mRNA expression and protein level were reduced in the islets from type-2 diabetic donors. RNA sequencing in human islets revealed GPRC5C expression correlated with the expression of genes controlling apoptosis, cell survival and proliferation. A reduction in Gprc5c mRNA and protein expression was observed in islets isolated from old mice (&gt;46 weeks of age) compared to that in islets from newborn (&lt;3 weeks) mice. Down-regulation of Gprc5c led to both moderately reduced glucose-stimulated insulin release and also reduced cAMP content in mouse islets. Potentiation of glucose-stimulated insulin secretion concomitant with enhanced islet cAMP level by all-trans retinoic acid (ATRA) was attenuated upon Gprc5c-KD. ATRA also increased [Ca<sup>+2</sup> ]<sub>i</sub> in Huh7-cells. Gprc5c over expression in Huh7 cells was associated with increased ERK1/2 activity. Gprc5c-KD in clonal MIN6c4 cells reduced cell proliferation and in murine islets increased apoptosis and the sensitivity of primary islet cells to a cocktail of pro-apoptotic cytokines. Our results demonstrate that agents activating GPRC5C represent a novel modality for the treatment and/or prevention of diabetes by restoring and/or maintaining functional β-cell mass.</p>},
  author       = {Amisten, Stefan and Al-Amily, Israa Mohammad and Soni, Arvind and Hawkes, Ross and Atanes, Patricio and Persaud, Shanta Jean and Rorsman, Patrik and Salehi, S Albert},
  issn         = {0918-8959},
  keyword      = {Diabetes,Insulin release,Orphan GPCR,RAIG2,RAIG3},
  language     = {eng},
  number       = {3},
  pages        = {325--338},
  publisher    = {Japan Endocrine Society},
  series       = {Endocrine Journal},
  title        = {Anti-diabetic action of all-trans retinoic acid and the orphan G protein coupled receptor GPRC5C in pancreatic β-cells},
  url          = {http://dx.doi.org/10.1507/endocrj.EJ16-0338},
  volume       = {64},
  year         = {2017},
}