RIPK1 and RIPK3 regulate TNFα-induced β-cell death in concert with caspase activity
Objective: Type 1 diabetes (T1D) is characterized by autoimmune-associated β-cell loss, insulin insufficiency, and hyperglycemia. Although TNFα signaling is associated with β-cell loss and hyperglycemia in non-obese diabetic mice and human T1D, the molecular mechanisms of β-cell TNF receptor signali...
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| Format: | Article |
| Language: | English |
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Elsevier
2022-11-01
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| Series: | Molecular Metabolism |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S221287782200151X |
| _version_ | 1817969195823923200 |
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| author | Christopher J. Contreras Noyonika Mukherjee Renato C.S. Branco Li Lin Meghan F. Hogan Erica P. Cai Andrew A. Oberst Steven E. Kahn Andrew T. Templin |
| author_facet | Christopher J. Contreras Noyonika Mukherjee Renato C.S. Branco Li Lin Meghan F. Hogan Erica P. Cai Andrew A. Oberst Steven E. Kahn Andrew T. Templin |
| author_sort | Christopher J. Contreras |
| collection | DOAJ |
| description | Objective: Type 1 diabetes (T1D) is characterized by autoimmune-associated β-cell loss, insulin insufficiency, and hyperglycemia. Although TNFα signaling is associated with β-cell loss and hyperglycemia in non-obese diabetic mice and human T1D, the molecular mechanisms of β-cell TNF receptor signaling have not been fully characterized. Based on work in other cell types, we hypothesized that receptor interacting protein kinase 1 (RIPK1) and receptor interacting protein kinase 3 (RIPK3) regulate TNFα-induced β-cell death in concert with caspase activity. Methods: We evaluated TNFα-induced cell death, caspase activity, and TNF receptor pathway molecule expression in immortalized NIT-1 and INS-1 β-cell lines and primary mouse islet cells in vitro. Our studies utilized genetic and small molecule approaches to alter RIPK1 and RIPK3 expression and caspase activity to interrogate mechanisms of TNFα-induced β-cell death. We used the β-cell toxin streptozotocin (STZ) to determine the susceptibility of Ripk3+/+ and Ripk3−/− mice to hyperglycemia in vivo. Results: Expression of TNF receptor signaling molecules including RIPK1 and RIPK3 was identified in NIT-1 and INS-1 β cells and isolated mouse islets at the mRNA and protein levels. TNFα treatment increased NIT-1 and INS-1 cell death and caspase activity after 24–48 h, and BV6, a small molecule inhibitor of inhibitor of apoptosis proteins (IAPs) amplified this TNFα-induced cell death. RIPK1 deficient NIT-1 cells were protected from TNFα- and BV6-induced cell death and caspase activation. Interestingly, small molecule inhibition of caspases with zVAD-fmk (zVAD) did not prevent TNFα-induced cell death in either NIT-1 or INS-1 cells. This caspase-independent cell death was increased by BV6 treatment and decreased in RIPK1 deficient NIT-1 cells. RIPK3 deficient NIT-1 cells and RIPK3 kinase inhibitor treated INS-1 cells were protected from TNFα+zVAD-induced cell death, whereas RIPK3 overexpression increased INS-1 cell death and promoted RIPK3 and MLKL interaction under TNFα+zVAD treatment. In mouse islet cells, BV6 or zVAD treatment promoted TNFα-induced cell death, and TNFα+zVAD-induced cell death was blocked by RIPK3 inhibition and in Ripk3−/− islet cells in vitro. Ripk3−/− mice were also protected from STZ-induced hyperglycemia and glucose intolerance in vivo. Conclusions: RIPK1 and RIPK3 regulate TNFα-induced β-cell death in concert with caspase activity in immortalized and primary islet β cells. TNF receptor signaling molecules such as RIPK1 and RIPK3 may represent novel therapeutic targets to promote β-cell survival and glucose homeostasis in T1D. |
| first_indexed | 2024-04-13T20:18:16Z |
| format | Article |
| id | doaj.art-7d60d82919e94293aee44d1fed96ab69 |
| institution | Directory Open Access Journal |
| issn | 2212-8778 |
| language | English |
| last_indexed | 2024-04-13T20:18:16Z |
| publishDate | 2022-11-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Metabolism |
| spelling | doaj.art-7d60d82919e94293aee44d1fed96ab692022-12-22T02:31:38ZengElsevierMolecular Metabolism2212-87782022-11-0165101582RIPK1 and RIPK3 regulate TNFα-induced β-cell death in concert with caspase activityChristopher J. Contreras0Noyonika Mukherjee1Renato C.S. Branco2Li Lin3Meghan F. Hogan4Erica P. Cai5Andrew A. Oberst6Steven E. Kahn7Andrew T. Templin8Division of Endocrinology, Department of Medicine, Roudebush VA Medical Center and Indiana University School of Medicine, Indianapolis, IN, USADepartment of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, IN, USALilly Diabetes Center of Excellence, Indiana Biosciences Research Institute, Indianapolis, IN, USALilly Diabetes Center of Excellence, Indiana Biosciences Research Institute, Indianapolis, IN, USADivision of Metabolism, Endocrinology and Nutrition, Department of Medicine, VA Puget Sound Health Care System and University of Washington, Seattle, WA, USALilly Diabetes Center of Excellence, Indiana Biosciences Research Institute, Indianapolis, IN, USADepartment of Immunology, University of Washington, Seattle, WA, USADivision of Metabolism, Endocrinology and Nutrition, Department of Medicine, VA Puget Sound Health Care System and University of Washington, Seattle, WA, USADivision of Endocrinology, Department of Medicine, Roudebush VA Medical Center and Indiana University School of Medicine, Indianapolis, IN, USA; Department of Biochemistry and Molecular Biology, 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; Corresponding author. 1210 Waterway Blvd., Ste. 2000, Indianapolis, IN, USA, 46202.Objective: Type 1 diabetes (T1D) is characterized by autoimmune-associated β-cell loss, insulin insufficiency, and hyperglycemia. Although TNFα signaling is associated with β-cell loss and hyperglycemia in non-obese diabetic mice and human T1D, the molecular mechanisms of β-cell TNF receptor signaling have not been fully characterized. Based on work in other cell types, we hypothesized that receptor interacting protein kinase 1 (RIPK1) and receptor interacting protein kinase 3 (RIPK3) regulate TNFα-induced β-cell death in concert with caspase activity. Methods: We evaluated TNFα-induced cell death, caspase activity, and TNF receptor pathway molecule expression in immortalized NIT-1 and INS-1 β-cell lines and primary mouse islet cells in vitro. Our studies utilized genetic and small molecule approaches to alter RIPK1 and RIPK3 expression and caspase activity to interrogate mechanisms of TNFα-induced β-cell death. We used the β-cell toxin streptozotocin (STZ) to determine the susceptibility of Ripk3+/+ and Ripk3−/− mice to hyperglycemia in vivo. Results: Expression of TNF receptor signaling molecules including RIPK1 and RIPK3 was identified in NIT-1 and INS-1 β cells and isolated mouse islets at the mRNA and protein levels. TNFα treatment increased NIT-1 and INS-1 cell death and caspase activity after 24–48 h, and BV6, a small molecule inhibitor of inhibitor of apoptosis proteins (IAPs) amplified this TNFα-induced cell death. RIPK1 deficient NIT-1 cells were protected from TNFα- and BV6-induced cell death and caspase activation. Interestingly, small molecule inhibition of caspases with zVAD-fmk (zVAD) did not prevent TNFα-induced cell death in either NIT-1 or INS-1 cells. This caspase-independent cell death was increased by BV6 treatment and decreased in RIPK1 deficient NIT-1 cells. RIPK3 deficient NIT-1 cells and RIPK3 kinase inhibitor treated INS-1 cells were protected from TNFα+zVAD-induced cell death, whereas RIPK3 overexpression increased INS-1 cell death and promoted RIPK3 and MLKL interaction under TNFα+zVAD treatment. In mouse islet cells, BV6 or zVAD treatment promoted TNFα-induced cell death, and TNFα+zVAD-induced cell death was blocked by RIPK3 inhibition and in Ripk3−/− islet cells in vitro. Ripk3−/− mice were also protected from STZ-induced hyperglycemia and glucose intolerance in vivo. Conclusions: RIPK1 and RIPK3 regulate TNFα-induced β-cell death in concert with caspase activity in immortalized and primary islet β cells. TNF receptor signaling molecules such as RIPK1 and RIPK3 may represent novel therapeutic targets to promote β-cell survival and glucose homeostasis in T1D.http://www.sciencedirect.com/science/article/pii/S221287782200151XRIPK1RIPK3β-cell deathType 1 diabetesTNFαCaspase |
| spellingShingle | Christopher J. Contreras Noyonika Mukherjee Renato C.S. Branco Li Lin Meghan F. Hogan Erica P. Cai Andrew A. Oberst Steven E. Kahn Andrew T. Templin RIPK1 and RIPK3 regulate TNFα-induced β-cell death in concert with caspase activity Molecular Metabolism RIPK1 RIPK3 β-cell death Type 1 diabetes TNFα Caspase |
| title | RIPK1 and RIPK3 regulate TNFα-induced β-cell death in concert with caspase activity |
| title_full | RIPK1 and RIPK3 regulate TNFα-induced β-cell death in concert with caspase activity |
| title_fullStr | RIPK1 and RIPK3 regulate TNFα-induced β-cell death in concert with caspase activity |
| title_full_unstemmed | RIPK1 and RIPK3 regulate TNFα-induced β-cell death in concert with caspase activity |
| title_short | RIPK1 and RIPK3 regulate TNFα-induced β-cell death in concert with caspase activity |
| title_sort | ripk1 and ripk3 regulate tnfα induced β cell death in concert with caspase activity |
| topic | RIPK1 RIPK3 β-cell death Type 1 diabetes TNFα Caspase |
| url | http://www.sciencedirect.com/science/article/pii/S221287782200151X |
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