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The transcriptional factor, Tead1, mediates the transcriptional output of the Hippo pathway. Tead1 activates the transcription of downstream genes including many cell cycle promoting genes that lead to cell proliferation in many tissues, but its role in β-cells is unknown. To test if Tead1 is required for β-cell proliferation and function, we deleted Tead1 in β-cell. Deletion of Tead1 in β-cells was confirmed in these β-cell Tead1 KO (β-Tead1-/-) , by a significant decrease in transcript and protein from whole islet lysates. The β-Tead1-/- mice had normal body weight but developed significantly higher fasting and fed glucose levels starting at 5 weeks of age and progressed to frank diabetes by 8 weeks, with fasting blood glucose >300 mg/dl, accompanied by hypoinsulinemia, as compared to floxed control mice. 8wk β-Tead1-/- mice display severe glucose intolerance due to an abrogation of GSIS during GTT, with a loss of first phase insulin secretion. Pancreatic insulin content was decreased by ∼50%, due to a decrease in expression of mature β-cell genes including, Pdx1, Nkx6.1 and MafA, all of which, we show to be direct transcriptional targets of Tead1. Tead1 also binds to the proximal promoter at the Ink4a locus to regulate p16/p19, critical cell cycle inhibitors, and this loss of activation in Tead1-deficient β-cells leads to an enhanced entry into cell cycle. Furthermore, to test if Tead1 has a similar regulatory role in human β-cells, we deleted TEAD1 knockout in human iPS cell-derived β-cells using CRISPR/Cas9 system. Tead1 deficient iPS cell-derived β-cells displayed a significant decrease in vitro GSIS. Our results demonstrate Tead1 to be a transcriptional switch required for β-cells to maintain mature function and remain in proliferative quiescence. This novel regulatory pathway could be targeted for β-cell replacement therapy to achieve β-cell proliferation without a loss of mature function. Disclosure J. Lee: None. V. Yechoor: n/a. R. Liu: None. F. Li: None. V. Negi: None. R. Jagannathan: None. M. Huising: Consultant; Crinetics Pharmaceuticals, Inc., Research Support; Crinetics Pharmaceuticals, Inc. K. Ma: None. B. Shih: None. M. Moulik: None. Funding VA-MERIT Grant (I01BX002678) |
