PMID- 38218538 OWN - NLM STAT- MEDLINE DCOM- 20240214 LR - 20240316 IS - 2212-8778 (Electronic) IS - 2212-8778 (Linking) VI - 80 DP - 2024 Feb TI - RIPK3 promotes islet amyloid-induced beta-cell loss and glucose intolerance in a humanized mouse model of type 2 diabetes. PG - 101877 LID - S2212-8778(24)00008-5 [pii] LID - 10.1016/j.molmet.2024.101877 [doi] LID - 101877 AB - OBJECTIVE: Aggregation of human islet amyloid polypeptide (hIAPP), a beta-cell secretory product, leads to islet amyloid deposition, islet inflammation and beta-cell loss in type 2 diabetes (T2D), but the mechanisms that underlie this process are incompletely understood. Receptor interacting protein kinase 3 (RIPK3) is a pro-death signaling molecule that has recently been implicated in amyloid-associated brain pathology and beta-cell cytotoxicity. Here, we evaluated the role of RIPK3 in amyloid-induced beta-cell loss using a humanized mouse model of T2D that expresses hIAPP and is prone to islet amyloid formation. METHODS: We quantified amyloid deposition, cell death and caspase 3/7 activity in islets isolated from WT, Ripk3(-/-), hIAPP and hIAPP; Ripk3(-/-) mice in real time, and evaluated hIAPP-stimulated inflammation in WT and Ripk3(-/-) bone marrow derived macrophages (BMDMs) in vitro. We also characterized the role of RIPK3 in glucose stimulated insulin secretion (GSIS) in vitro and in vivo. Finally, we examined the role of RIPK3 in high fat diet (HFD)-induced islet amyloid deposition, beta-cell loss and glucose homeostasis in vivo. RESULTS: We found that amyloid-prone hIAPP mouse islets exhibited increased cell death and caspase 3/7 activity compared to amyloid-free WT islets in vitro, and this was associated with increased RIPK3 expression. hIAPP; Ripk3(-/-) islets were protected from amyloid-induced cell death compared to hIAPP islets in vitro, although amyloid deposition and caspase 3/7 activity were not different between genotypes. We observed that macrophages are a source of Ripk3 expression in isolated islets, and that Ripk3(-/-) BMDMs were protected from hIAPP-stimulated inflammatory gene expression (Tnf, Il1b, Nos2). Following 52 weeks of HFD feeding, islet amyloid-prone hIAPP mice exhibited impaired glucose tolerance and decreased beta-cell area compared to WT mice in vivo, whereas hIAPP; Ripk3(-/-) mice were protected from these impairments. CONCLUSIONS: In conclusion, loss of RIPK3 protects from amyloid-induced inflammation and islet cell death in vitro and amyloid-induced beta-cell loss and glucose intolerance in vivo. We propose that therapies targeting RIPK3 may reduce islet inflammation and beta-cell loss and improve glucose homeostasis in the pathogenesis of T2D. CI - Published by Elsevier GmbH. FAU - Mukherjee, Noyonika AU - Mukherjee N AD - Department of Biochemistry & Molecular Biology, Indiana University School of Medicine, Indianapolis, IN, USA. FAU - Contreras, Christopher J AU - Contreras CJ AD - Division of Endocrinology, Department of Medicine, Roudebush VA Medical Center and Indiana University School of Medicine, Indianapolis, IN, USA. FAU - Lin, Li AU - Lin L AD - Lilly Diabetes Center of Excellence, Indiana Biosciences Research Institute, Indianapolis, IN, USA. FAU - Colglazier, Kaitlyn A AU - Colglazier KA AD - Lilly Diabetes Center of Excellence, Indiana Biosciences Research Institute, Indianapolis, IN, USA. FAU - Mather, Egan G AU - Mather EG AD - Lilly Diabetes Center of Excellence, Indiana Biosciences Research Institute, Indianapolis, IN, USA. FAU - Kalwat, Michael A AU - Kalwat MA AD - Lilly Diabetes Center of Excellence, Indiana Biosciences Research Institute, Indianapolis, IN, USA. FAU - Esser, Nathalie AU - Esser N AD - Division of Metabolism, Endocrinology and Nutrition, Department of Medicine, VA Puget Sound Health Care System and the University of Washington, Seattle, WA, USA. FAU - Kahn, Steven E AU - Kahn SE AD - Division of Metabolism, Endocrinology and Nutrition, Department of Medicine, VA Puget Sound Health Care System and the University of Washington, Seattle, WA, USA. FAU - Templin, Andrew T AU - Templin AT AD - Department of Biochemistry & Molecular Biology, Indiana University School of Medicine, Indianapolis, IN, USA; Division of Endocrinology, Department of Medicine, Roudebush VA Medical Center and Indiana University School of Medicine, Indianapolis, IN, USA; Lilly Diabetes Center of Excellence, Indiana Biosciences Research Institute, Indianapolis, IN, USA; Center for Diabetes and Metabolic Diseases, Indiana University School of Medicine, Indianapolis, IN, USA. Electronic address: templin@iu.edu. LA - eng GR - T32 DK007247/DK/NIDDK NIH HHS/United States PT - Journal Article DEP - 20240111 PL - Germany TA - Mol Metab JT - Molecular metabolism JID - 101605730 RN - 0 (Amyloid) RN - 0 (Amyloid beta-Peptides) RN - EC 3.4.22.- (Caspase 3) RN - IY9XDZ35W2 (Glucose) RN - 0 (Islet Amyloid Polypeptide) RN - EC 2.7.11.1 (Receptor-Interacting Protein Serine-Threonine Kinases) RN - EC 2.7.11.1 (Ripk3 protein, mouse) SB - IM MH - Animals MH - Humans MH - Mice MH - Amyloid/metabolism MH - Amyloid beta-Peptides/metabolism MH - Caspase 3/metabolism MH - *Diabetes Mellitus, Type 2/metabolism MH - Glucose MH - *Glucose Intolerance MH - Inflammation MH - Islet Amyloid Polypeptide/genetics/metabolism MH - *Receptor-Interacting Protein Serine-Threonine Kinases/genetics PMC - PMC10830894 OTO - NOTNLM OT - Diabetes OT - Islet amyloid OT - RIPK3 OT - beta-cell cytotoxicity COIS- Declaration of competing interest All authors declare that no competing interests exist. EDAT- 2024/01/14 12:42 MHDA- 2024/02/10 10:43 PMCR- 2024/01/11 CRDT- 2024/01/13 19:34 PHST- 2023/11/06 00:00 [received] PHST- 2023/12/29 00:00 [revised] PHST- 2024/01/09 00:00 [accepted] PHST- 2024/02/10 10:43 [medline] PHST- 2024/01/14 12:42 [pubmed] PHST- 2024/01/13 19:34 [entrez] PHST- 2024/01/11 00:00 [pmc-release] AID - S2212-8778(24)00008-5 [pii] AID - 101877 [pii] AID - 10.1016/j.molmet.2024.101877 [doi] PST - ppublish SO - Mol Metab. 2024 Feb;80:101877. doi: 10.1016/j.molmet.2024.101877. Epub 2024 Jan 11.