Quantitative-proteomic comparison of alpha and Beta cells to uncover novel targets for lineage reprogramming.
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.
Society of Fellows, Harvard University, Cambridge, Massachusetts, United States of America; Center for the Science of Therapeutics, Broad Institute, Cambridge, Massachusetts, United States of America.
Amit
A
Choudhary
Center for the Science of Therapeutics, Broad Institute, Cambridge, Massachusetts, United States of America.
Kaihui
K
Hu He
Center for the Science of Therapeutics, Broad Institute, Cambridge, Massachusetts, United States of America.
Philipp
P
Mertins
Center for the Science of Therapeutics, Broad Institute, Cambridge, Massachusetts, United States of America.
Namrata D
ND
Udeshi
Center for the Science of Therapeutics, Broad Institute, Cambridge, Massachusetts, United States of America.
Vlado
V
Dančík
Center for the Science of Therapeutics, Broad Institute, Cambridge, Massachusetts, United States of America; Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, United States of America.
Dina
D
Fomina-Yadlin
Center for the Science of Therapeutics, Broad Institute, Cambridge, Massachusetts, United States of America.
Stefan
S
Kubicek
Center for the Science of Therapeutics, Broad Institute, Cambridge, Massachusetts, United States of America.
Paul A
PA
Clemons
Center for the Science of Therapeutics, Broad Institute, Cambridge, Massachusetts, United States of America; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, United States of America; Howard Hughes Medical Institute, Chevy Chase, Maryland, United States of America.
Stuart L
SL
Schreiber
Center for the Science of Therapeutics, Broad Institute, Cambridge, Massachusetts, United States of America.
Steven A
SA
Carr
Center for the Science of Therapeutics, Broad Institute, Cambridge, Massachusetts, United States of America.
Bridget K
BK
Wagner
D000818:Animals; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase / Q000378:metabolism; D002460:Cell Line; D050416:Glucagon-Secreting Cells / Q000378:metabolism*; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D051379:Mice; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D040901:Proteomics / Q000379:methods*
D047908:Intracellular Signaling Peptides and Proteins; C523084:BRSK1 protein, human; D017346:Protein Serine-Threonine Kinases; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase; C517700:Camkk2 protein, mouse