Quantitative-proteomic comparison of alpha and Beta cells to uncover novel targets for lineage reprogramming

PLoS One. 2014 Apr 23;9(4):e95194. doi: 10.1371/journal.pone.0095194. eCollection 2014.

Abstract

Type-1 diabetes (T1D) is an autoimmune disease in which insulin-secreting pancreatic beta cells are destroyed by the immune system. An emerging strategy to regenerate beta-cell mass is through transdifferentiation of pancreatic alpha cells to beta cells. We previously reported two small molecules, BRD7389 and GW8510, that induce insulin expression in a mouse alpha cell line and provide a glimpse into potential intermediate cell states in beta-cell reprogramming from alpha cells. These small-molecule studies suggested that inhibition of kinases in particular may induce the expression of several beta-cell markers in alpha cells. To identify potential lineage reprogramming protein targets, we compared the transcriptome, proteome, and phosphoproteome of alpha cells, beta cells, and compound-treated alpha cells. Our phosphoproteomic analysis indicated that two kinases, BRSK1 and CAMKK2, exhibit decreased phosphorylation in beta cells compared to alpha cells, and in compound-treated alpha cells compared to DMSO-treated alpha cells. Knock-down of these kinases in alpha cells resulted in expression of key beta-cell markers. These results provide evidence that perturbation of the kinome may be important for lineage reprogramming of alpha cells to beta cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium-Calmodulin-Dependent Protein Kinase Kinase / metabolism
  • Cell Line
  • Glucagon-Secreting Cells / metabolism*
  • Insulin-Secreting Cells / metabolism*
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Mice
  • Phosphorylation
  • Protein-Serine-Threonine Kinases / metabolism
  • Proteomics / methods*

Substances

  • Intracellular Signaling Peptides and Proteins
  • BRSK1 protein, human
  • Protein-Serine-Threonine Kinases
  • Calcium-Calmodulin-Dependent Protein Kinase Kinase
  • Camkk2 protein, mouse

Grant support

This work was supported by JDRF 17-2008-1030 and 17-2011-260 (BKW, SLS) and Harvard Society of Fellows (AC). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.