Role of poly(ADP-ribose) polymerase-1 in regulating human islet cell differentiation
Abstract Poly(ADP-ribose) polymerase-1 (PARP1), a fundamental DNA repair enzyme, is known to regulate β cell death, replication, and insulin secretion. PARP1 knockout (KO) mice are resistant to diabetes, while PARP1 overactivation contributes to β cell death. Additionally, PARP1 inhibition (PARPi) i...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-12-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-25405-w |
| _version_ | 1797980305532911616 |
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| author | Nidheesh Dadheech Abhay Srivastava Rashmi G. Shah Girish M. Shah Sarita Gupta |
| author_facet | Nidheesh Dadheech Abhay Srivastava Rashmi G. Shah Girish M. Shah Sarita Gupta |
| author_sort | Nidheesh Dadheech |
| collection | DOAJ |
| description | Abstract Poly(ADP-ribose) polymerase-1 (PARP1), a fundamental DNA repair enzyme, is known to regulate β cell death, replication, and insulin secretion. PARP1 knockout (KO) mice are resistant to diabetes, while PARP1 overactivation contributes to β cell death. Additionally, PARP1 inhibition (PARPi) improves diabetes complications in patients with type-2 diabetes. Despite these beneficial effects, the use of PARP1 modulating agents in diabetes treatment is largely neglected, primarily due to the poorly studied mechanistic action of PARP1 catalytic function in human β cell development. In the present study, we evaluated PARP1 regulatory action in human β cell differentiation using the human pancreatic progenitor cell line, PANC-1. We surveyed islet census and histology from PARP1 wild-type versus KO mice pancreas in a head-to-head comparison with PARP1 regulatory action for in-vitro β cell differentiation following either PARP1 depletion or its pharmacological inhibition in PANC-1-differentiated islet cells. shRNA mediated PARP1 depleted (SiP) and shRNA control (U6) PANC-1 cells were differentiated into islet-like clusters using established protocols. We observed complete abrogation of new β cell formation with absolute PARP1 depletion while its inhibition using the potent inhibitor, PJ34, promoted the endocrine β cell differentiation and maturation. Immunohistochemistry and immunoblotting for key endocrine differentiation players along with β cell maturation markers highlighted the potential regulatory action of PARP1 and augmented β cell differentiation due to direct interaction of unmodified PARP1 protein elicited p38 MAPK phosphorylation and Neurogenin-3 (Ngn3) re-activation. In summary, our study suggests that PARP1 is required for the proper development and differentiation of human islets. Selective inhibition with PARPi can be an advantage in pushing more insulin-producing cells under pathological conditions and delivers a potential for pilot clinical testing for β cell replacement cell therapies for diabetes. |
| first_indexed | 2024-04-11T05:53:29Z |
| format | Article |
| id | doaj.art-bc26e8588226487496c13f071d0f0c6a |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-11T05:53:29Z |
| publishDate | 2022-12-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-bc26e8588226487496c13f071d0f0c6a2022-12-22T04:42:00ZengNature PortfolioScientific Reports2045-23222022-12-0112111610.1038/s41598-022-25405-wRole of poly(ADP-ribose) polymerase-1 in regulating human islet cell differentiationNidheesh Dadheech0Abhay Srivastava1Rashmi G. Shah2Girish M. Shah3Sarita Gupta4Department of Surgery, Alberta Diabetes Institute, University of AlbertaDepartment of Biochemistry, Faculty of Science, The Maharaja Sayajirao University of BarodaCHU de Quebec Laval University Hospital Research Centres Quebec CityCHU de Quebec Laval University Hospital Research Centres Quebec CityDepartment of Biochemistry, Faculty of Science, The Maharaja Sayajirao University of BarodaAbstract Poly(ADP-ribose) polymerase-1 (PARP1), a fundamental DNA repair enzyme, is known to regulate β cell death, replication, and insulin secretion. PARP1 knockout (KO) mice are resistant to diabetes, while PARP1 overactivation contributes to β cell death. Additionally, PARP1 inhibition (PARPi) improves diabetes complications in patients with type-2 diabetes. Despite these beneficial effects, the use of PARP1 modulating agents in diabetes treatment is largely neglected, primarily due to the poorly studied mechanistic action of PARP1 catalytic function in human β cell development. In the present study, we evaluated PARP1 regulatory action in human β cell differentiation using the human pancreatic progenitor cell line, PANC-1. We surveyed islet census and histology from PARP1 wild-type versus KO mice pancreas in a head-to-head comparison with PARP1 regulatory action for in-vitro β cell differentiation following either PARP1 depletion or its pharmacological inhibition in PANC-1-differentiated islet cells. shRNA mediated PARP1 depleted (SiP) and shRNA control (U6) PANC-1 cells were differentiated into islet-like clusters using established protocols. We observed complete abrogation of new β cell formation with absolute PARP1 depletion while its inhibition using the potent inhibitor, PJ34, promoted the endocrine β cell differentiation and maturation. Immunohistochemistry and immunoblotting for key endocrine differentiation players along with β cell maturation markers highlighted the potential regulatory action of PARP1 and augmented β cell differentiation due to direct interaction of unmodified PARP1 protein elicited p38 MAPK phosphorylation and Neurogenin-3 (Ngn3) re-activation. In summary, our study suggests that PARP1 is required for the proper development and differentiation of human islets. Selective inhibition with PARPi can be an advantage in pushing more insulin-producing cells under pathological conditions and delivers a potential for pilot clinical testing for β cell replacement cell therapies for diabetes.https://doi.org/10.1038/s41598-022-25405-w |
| spellingShingle | Nidheesh Dadheech Abhay Srivastava Rashmi G. Shah Girish M. Shah Sarita Gupta Role of poly(ADP-ribose) polymerase-1 in regulating human islet cell differentiation Scientific Reports |
| title | Role of poly(ADP-ribose) polymerase-1 in regulating human islet cell differentiation |
| title_full | Role of poly(ADP-ribose) polymerase-1 in regulating human islet cell differentiation |
| title_fullStr | Role of poly(ADP-ribose) polymerase-1 in regulating human islet cell differentiation |
| title_full_unstemmed | Role of poly(ADP-ribose) polymerase-1 in regulating human islet cell differentiation |
| title_short | Role of poly(ADP-ribose) polymerase-1 in regulating human islet cell differentiation |
| title_sort | role of poly adp ribose polymerase 1 in regulating human islet cell differentiation |
| url | https://doi.org/10.1038/s41598-022-25405-w |
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