Dual role of proapoptotic BAD in insulin secretion and beta cell survival

Abstract

The proapoptotic BCL-2 family member BAD resides in a glucokinase-containing complex that regulates glucose-driven mitochondrial respiration. Here, we present genetic evidence of a physiologic role for BAD in glucose-stimulated insulin secretion by beta cells. This novel function of BAD is specifically dependent upon the phosphorylation of its BH3 sequence, previously defined as an essential death domain. We highlight the pharmacologic relevance of phosphorylated BAD BH3 by using cell-permeable, hydrocarbon-stapled BAD BH3 helices that target glucokinase, restore glucose-driven mitochondrial respiration and correct the insulin secretory response in Bad-deficient islets. Our studies uncover an alternative target and function for the BAD BH3 domain and emphasize the therapeutic potential of phosphorylated BAD BH3 mimetics in selectively restoring beta cell function. Furthermore, we show that BAD regulates the physiologic adaptation of beta cell mass during high-fat feeding. Our findings provide genetic proof of the bifunctional activities of BAD in both beta cell survival and insulin secretion.

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Figure 1: Impaired insulin secretion in Bad−/− mice.
Figure 2: Characterization of the insulin secretion defect in Bad−/− islets.
Figure 3: Glucose-induced changes in mitochondrial membrane potential and [Ca2+]i in Bad+/+ and Bad−/− beta cells.
Figure 4: Regulation of GSIS by the BAD BH3 domain and its phosphorylation status.
Figure 5: Metabolic activity of SAHB compounds in beta cells.
Figure 6: Sensitivity of Bad genetic models to HFD.

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Acknowledgements

We thank M. Godes and C. Gramm for technical assistance, R. Pasquier and B. Szlyk for islet isolation and animal husbandry, G. Weir (Joslin Diabetes Center, Boston, MA) for antibody to glucokinase, B. Spiegelman, B. Malynn, M. Vander Heiden, A. Schinzel, J. Labelle, G. Verdine, F. Bernal, A. Saghatelian, A.-M. Richards and G. Yaney for helpful discussion and E. Smith for manuscript preparation. N.N.D. and L.D.W. are recipients of the Burroughs Wellcome Fund Career Award in Biomedical Sciences. This work was supported by US National Institute of Health grants K01CA10659 (N.N.D.), 5R01CA50239 and 5R01DK68781 (S.J.K.), 5K08HL074049 (L.D.W.) and by Charles E. Culpeper Scholarship in Medical Science (L.D.W.).

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N.N.D. planned and performed the in vivo and in vitro experiments, supervised the project and wrote the manuscript. J.K.F. and K.R. assisted with conducting experiments. L.D.W., K.L.P., G.H.B. and J.M. designed and synthesized the SAHB compounds and characterized their chemical and biophysical properties. C.-Y.Z. made conceptual contributions and helped with data interpretation. C.S.C. performed the hyperglycemic clamp studies with help from A.K., S.N., S.K. and supervision from G.I.S. A.J.A.M. and J.D.W. performed the measurements of mitochondrial membrane potential with supervision from O.S.S. S.R.D. generated the BadS155A knock-in model with supervision from M.E.G. J.T.D. helped with islet perifusion studies. B.E.C. provided critical guidance and expertise. B.B.L. provided advice. The late S.J.K. provided the mentorship and laboratory resources that catalyzed the initial studies.

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Correspondence to Nika N Danial.

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Supplementary Figs. 1–4, Supplementary Table 1 and Supplementary Methods (PDF 645 kb)

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Danial, N., Walensky, L., Zhang, CY. et al. Dual role of proapoptotic BAD in insulin secretion and beta cell survival. Nat Med 14, 144–153 (2008). https://doi.org/10.1038/nm1717

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