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Updated: Apr 30, 2026
In Vitro Colony Assays for Characterizing Tri-potent Progenitor Cells Isolated from the Adult Murine Pancreas
Published on: June 10, 2016
Quantitative-proteomic comparison of alpha and Beta cells to uncover novel targets for lineage reprogramming
Amit Choudhary1, Kaihui Hu He2, Philipp Mertins2
1Society of Fellows, Harvard University, Cambridge, Massachusetts, United States of America; Center for the Science of Therapeutics, Broad Institute, Cambridge, Massachusetts, United States of America.
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.
Insights
Type-1 diabetes (T1D) research explores regenerating insulin-producing beta cells. Inhibiting specific kinases (BRSK1, CAMKK2) in alpha cells promotes beta-cell marker expression, offering a new reprogramming strategy.
Area of Science:
- Cell biology
- Endocrinology
- Immunology
Background:
- Type-1 diabetes (T1D) involves the immune system destroying insulin-producing beta cells.
- Regenerating beta-cell mass via alpha-to-beta cell transdifferentiation is a promising therapeutic strategy.
- Previous studies identified small molecules that induce insulin expression in alpha cells, suggesting kinase inhibition's role.
Purpose of the Study:
- To identify protein targets for lineage reprogramming of alpha cells into beta cells.
- To investigate the role of kinase activity in alpha-to-beta cell transdifferentiation.
Main Methods:
- Comparative analysis of transcriptome, proteome, and phosphoproteome in alpha cells, beta cells, and compound-treated alpha cells.
- Phosphoproteomic analysis to identify differentially phosphorylated kinases.
- Knock-down experiments of identified kinases in alpha cells.
Main Results:
- Phosphoproteomic analysis revealed decreased phosphorylation of BRSK1 and CAMKK2 in beta cells and compound-treated alpha cells.
- Knock-down of BRSK1 and CAMKK2 in alpha cells led to the expression of key beta-cell markers.
- These findings highlight the kinome's importance in alpha-to-beta cell reprogramming.
Conclusions:
- Perturbation of the kinome, specifically BRSK1 and CAMKK2, is crucial for alpha-to-beta cell lineage reprogramming.
- Targeting these kinases offers a novel approach for T1D regenerative therapy.
- Further research into kinase pathways could unlock new strategies for beta-cell regeneration.
