{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T23:21:07Z","timestamp":1768519267604,"version":"3.49.0"},"posted":{"date-parts":[[2024,12,26]]},"group-title":"Cell Biology","reference-count":87,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2024,12,26]],"date-time":"2024-12-26T00:00:00Z","timestamp":1735171200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,12,26]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>Obesity, insulin resistance, and a host of environmental and genetic factors can drive hyperglycemia, causing \u03b2-cells to compensate by increasing insulin production and secretion. In type 2 diabetes (T2D), \u03b2-cells under these conditions eventually fail. Rare \u03b2-cell diseases like congenital hyperinsulinism (HI) also cause inappropriate insulin secretion, and some HI patients develop diabetes. However, the mechanisms of insulin hypersecretion and how it causes \u03b2-cell dysfunction are not fully understood. We previously discovered small molecules (e.g. SW016789) that cause insulin hypersecretion and lead to a loss in \u03b2-cell function without cell death. Here, we uncover the protein target of SW016789 and provide the first time-course transcriptomic analysis of hypersecretory responses versus thapsigargin-mediated ER stress in \u03b2-cells. In mouse MIN6 and human EndoC-\u03b2H1 \u03b2-cells, we identified and validated VDAC1 as a SW016789 target using photoaffinity proteomics, cellular thermal shift assays, siRNA, and small molecule inhibitors.<\/jats:p>\n                <jats:p>\n                  SW016789 raises membrane potential to enhance Ca\n                  <jats:sup>2+<\/jats:sup>\n                  influx, potentially through VDAC1. Chronically elevated intracellular Ca\n                  <jats:sup>2+<\/jats:sup>\n                  appears to underpin the negative impacts of hypersecretion, as nifedipine protected against each small molecule hypersecretion inducer we tested. Using time- course RNAseq, we discovered that hypersecretion induced a distinct transcriptional pattern compared to ER stress. Clustering analyses led us to focus on ER-associated degradation (ERAD) as a potential mediator of the adaptive response. SW016789 reduced the abundance of ERAD substrate OS-9 and pharmacological inhibition of ERAD worsened \u03b2-cell survival in response to hypersecretory stress. Changes in other ERAD components in MIN6 and EndoC-\u03b2H1 at the protein level were minor with either SW016789 or thapsigargin. However, immunostaining for core ERAD components SEL1L, HRD1, and DERL3 in non-diabetic and T2D human pancreas revealed altered distributions of SEL1L\/HRD1 and SEL1L\/DERL3 rations in \u03b2-cells of T2D islets, in alignment with altered ERAD in stressed \u03b2-cells. We conclude that hypersecretory stimuli, including SW016789- mediated VDAC1 activation, cause enhanced Ca\n                  <jats:sup>2+<\/jats:sup>\n                  influx and insulin release. Subsequent differential gene expression represents a \u03b2-cell hypersecretory response signature that is reflected at the protein level for some, but not all genes. A better understanding of how \u03b2-cells induce hypersecretion and the mechanisms of negative feedback on secretory rate may lead to the discovery of novel therapeutic targets for T2D and HI.\n                <\/jats:p>","DOI":"10.1101\/2024.12.26.629558","type":"posted-content","created":{"date-parts":[[2024,12,27]],"date-time":"2024-12-27T02:50:12Z","timestamp":1735267812000},"source":"Crossref","is-referenced-by-count":0,"title":["Target deconvolution of an insulin hypersecretion-inducer acting through VDAC1 with a distinct transcriptomic signature in beta-cells"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9622-0184","authenticated-orcid":false,"given":"Gitanjali","family":"Roy","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrea","family":"Ord\u00f3\u00f1ez","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4949-3507","authenticated-orcid":false,"given":"Derk D.","family":"Binns","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3012-7171","authenticated-orcid":false,"given":"Karina","family":"Rodrigues dos Santos","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael B.","family":"Kwakye","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-9809-0989","authenticated-orcid":false,"given":"George C.","family":"King","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6692-3631","authenticated-orcid":false,"given":"Noyonika","family":"Mukherjee","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6241-9304","authenticated-orcid":false,"given":"Andrew T.","family":"Templin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7696-113X","authenticated-orcid":false,"given":"Zhiyong","family":"Tan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1119-2752","authenticated-orcid":false,"given":"Timothy I.","family":"Richardson","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0049-0073","authenticated-orcid":false,"given":"Emma H.","family":"Doud","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5822-2894","authenticated-orcid":false,"given":"Amber L.","family":"Mosley","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9130-4194","authenticated-orcid":false,"given":"Christopher H.","family":"Emfinger","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0568-2261","authenticated-orcid":false,"given":"Alan D.","family":"Attie","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7405-5552","authenticated-orcid":false,"given":"Mark P.","family":"Keller","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4628-2256","authenticated-orcid":false,"given":"Travis S.","family":"Johnson","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8349-9470","authenticated-orcid":false,"given":"Michael A.","family":"Kalwat","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2024122716350519000_2024.12.26.629558v1.1","unstructured":"IDF Diabetes Atlas, 10th edn. Brussels. Belgium: International Diabetes Federation. 2021"},{"key":"2024122716350519000_2024.12.26.629558v1.2","doi-asserted-by":"publisher","DOI":"10.2337\/diabetes.54.11.3265"},{"key":"2024122716350519000_2024.12.26.629558v1.3","doi-asserted-by":"publisher","DOI":"10.1007\/s00424-020-02383-4"},{"key":"2024122716350519000_2024.12.26.629558v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2022.06.003"},{"key":"2024122716350519000_2024.12.26.629558v1.5","doi-asserted-by":"publisher","DOI":"10.1038\/312446a0"},{"key":"2024122716350519000_2024.12.26.629558v1.6","doi-asserted-by":"publisher","DOI":"10.1038\/311269a0"},{"key":"2024122716350519000_2024.12.26.629558v1.7","doi-asserted-by":"publisher","DOI":"10.1007\/BF00583760"},{"key":"2024122716350519000_2024.12.26.629558v1.8","doi-asserted-by":"publisher","DOI":"10.1007\/BF00585354"},{"key":"2024122716350519000_2024.12.26.629558v1.9","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00008.2017"},{"key":"2024122716350519000_2024.12.26.629558v1.10","doi-asserted-by":"publisher","DOI":"10.1002\/cphy.c210004"},{"key":"2024122716350519000_2024.12.26.629558v1.11","doi-asserted-by":"publisher","DOI":"10.2337\/diab.37.5.495"},{"key":"2024122716350519000_2024.12.26.629558v1.12","doi-asserted-by":"publisher","DOI":"10.2337\/diabetes.49.11.1751"},{"key":"2024122716350519000_2024.12.26.629558v1.13","doi-asserted-by":"publisher","DOI":"10.1007\/s00125-009-1314-y"},{"key":"2024122716350519000_2024.12.26.629558v1.14","doi-asserted-by":"publisher","DOI":"10.1210\/endocr\/bqab173"},{"key":"2024122716350519000_2024.12.26.629558v1.15","doi-asserted-by":"publisher","DOI":"10.2337\/dbi21-0009"},{"key":"2024122716350519000_2024.12.26.629558v1.16","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2022.12.012"},{"key":"2024122716350519000_2024.12.26.629558v1.17","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M111.254995"},{"key":"2024122716350519000_2024.12.26.629558v1.18","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2022.12.001"},{"key":"2024122716350519000_2024.12.26.629558v1.19","doi-asserted-by":"publisher","DOI":"10.2337\/db20-0377"},{"key":"2024122716350519000_2024.12.26.629558v1.20","doi-asserted-by":"publisher","DOI":"10.2337\/diabetes.49.12.2094"},{"key":"2024122716350519000_2024.12.26.629558v1.21","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pmed.0050206"},{"key":"2024122716350519000_2024.12.26.629558v1.22","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.adi8317"},{"key":"2024122716350519000_2024.12.26.629558v1.23","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2016.01.009"},{"key":"2024122716350519000_2024.12.26.629558v1.24","doi-asserted-by":"publisher","DOI":"10.1210\/endocr\/bqac081"},{"key":"2024122716350519000_2024.12.26.629558v1.25","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0258054"},{"key":"2024122716350519000_2024.12.26.629558v1.26","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2023.112615"},{"key":"2024122716350519000_2024.12.26.629558v1.27","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M203988200"},{"key":"2024122716350519000_2024.12.26.629558v1.28","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.abq4126"},{"key":"2024122716350519000_2024.12.26.629558v1.29","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2018.09.008"},{"key":"2024122716350519000_2024.12.26.629558v1.30","doi-asserted-by":"publisher","DOI":"10.1021\/acschembio.9b00399"},{"key":"2024122716350519000_2024.12.26.629558v1.31","doi-asserted-by":"publisher","DOI":"10.1370\/afm.3129"},{"key":"2024122716350519000_2024.12.26.629558v1.32","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-0716-1044-2_9"},{"key":"2024122716350519000_2024.12.26.629558v1.33","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1005896"},{"key":"2024122716350519000_2024.12.26.629558v1.34","doi-asserted-by":"publisher","DOI":"10.2337\/db10-0522"},{"key":"2024122716350519000_2024.12.26.629558v1.35","doi-asserted-by":"publisher","DOI":"10.1172\/JCI163584"},{"key":"2024122716350519000_2024.12.26.629558v1.36","doi-asserted-by":"publisher","DOI":"10.1172\/JCI134874"},{"key":"2024122716350519000_2024.12.26.629558v1.37","doi-asserted-by":"publisher","DOI":"10.2337\/db18-0624"},{"key":"2024122716350519000_2024.12.26.629558v1.38","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2008.11.045"},{"key":"2024122716350519000_2024.12.26.629558v1.39","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21176146"},{"key":"2024122716350519000_2024.12.26.629558v1.40","doi-asserted-by":"publisher","DOI":"10.1111\/bph.15018"},{"key":"2024122716350519000_2024.12.26.629558v1.41","doi-asserted-by":"publisher","DOI":"10.4161\/isl.2.5.12639"},{"key":"2024122716350519000_2024.12.26.629558v1.42","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.159301"},{"key":"2024122716350519000_2024.12.26.629558v1.43","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamem.2011.10.005"},{"key":"2024122716350519000_2024.12.26.629558v1.44","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0182027"},{"key":"2024122716350519000_2024.12.26.629558v1.45","doi-asserted-by":"publisher","DOI":"10.1016\/j.cellsig.2008.09.015"},{"key":"2024122716350519000_2024.12.26.629558v1.46","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.200608073"},{"key":"2024122716350519000_2024.12.26.629558v1.47","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2022.102035"},{"key":"2024122716350519000_2024.12.26.629558v1.48","doi-asserted-by":"publisher","DOI":"10.1186\/s40035-022-00329-7"},{"key":"2024122716350519000_2024.12.26.629558v1.49","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M116.744284"},{"key":"2024122716350519000_2024.12.26.629558v1.50","doi-asserted-by":"publisher","DOI":"10.2337\/db16-0084"},{"key":"2024122716350519000_2024.12.26.629558v1.51","doi-asserted-by":"publisher","DOI":"10.1210\/en.2010-1420"},{"key":"2024122716350519000_2024.12.26.629558v1.52","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2121-11-59"},{"key":"2024122716350519000_2024.12.26.629558v1.53","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2024.110291"},{"key":"2024122716350519000_2024.12.26.629558v1.54","doi-asserted-by":"publisher","DOI":"10.2337\/db12-1611"},{"key":"2024122716350519000_2024.12.26.629558v1.55","doi-asserted-by":"publisher","DOI":"10.2337\/db21-0755"},{"key":"2024122716350519000_2024.12.26.629558v1.56","doi-asserted-by":"publisher","DOI":"10.1055\/s-0029-1212000"},{"key":"2024122716350519000_2024.12.26.629558v1.57","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1096-9136(199804)15:4<297::AID-DIA572>3.0.CO;2-W"},{"key":"2024122716350519000_2024.12.26.629558v1.58","doi-asserted-by":"publisher","DOI":"10.2337\/diacare.17.1.45"},{"key":"2024122716350519000_2024.12.26.629558v1.59","doi-asserted-by":"publisher","DOI":"10.2337\/db24-0650"},{"key":"2024122716350519000_2024.12.26.629558v1.60","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M115.704098"},{"key":"2024122716350519000_2024.12.26.629558v1.61","doi-asserted-by":"publisher","DOI":"10.2337\/db12-1527"},{"key":"2024122716350519000_2024.12.26.629558v1.62","doi-asserted-by":"publisher","DOI":"10.1038\/nature19102"},{"key":"2024122716350519000_2024.12.26.629558v1.63","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2019.04.043"},{"key":"2024122716350519000_2024.12.26.629558v1.64","doi-asserted-by":"publisher","DOI":"10.1161\/01.RES.55.3.336"},{"key":"2024122716350519000_2024.12.26.629558v1.65","doi-asserted-by":"publisher","DOI":"10.2337\/db19-1060"},{"key":"2024122716350519000_2024.12.26.629558v1.66","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2004.09.035"},{"key":"2024122716350519000_2024.12.26.629558v1.67","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M117.785055"},{"key":"2024122716350519000_2024.12.26.629558v1.68","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0034298"},{"key":"2024122716350519000_2024.12.26.629558v1.69","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-024-49724-w"},{"key":"2024122716350519000_2024.12.26.629558v1.70","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2019.01.010"},{"key":"2024122716350519000_2024.12.26.629558v1.71","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0022713"},{"key":"2024122716350519000_2024.12.26.629558v1.72","doi-asserted-by":"publisher","DOI":"10.1021\/acssensors.6b00433"},{"key":"2024122716350519000_2024.12.26.629558v1.73","doi-asserted-by":"publisher","DOI":"10.3390\/cancers16020423"},{"key":"2024122716350519000_2024.12.26.629558v1.74","doi-asserted-by":"publisher","DOI":"10.3390\/proteomes9010015"},{"key":"2024122716350519000_2024.12.26.629558v1.75","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/bts635"},{"key":"2024122716350519000_2024.12.26.629558v1.76","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/bts731"},{"key":"2024122716350519000_2024.12.26.629558v1.77","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btt656"},{"key":"2024122716350519000_2024.12.26.629558v1.78","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1003118"},{"key":"2024122716350519000_2024.12.26.629558v1.79","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2105-9-559"},{"key":"2024122716350519000_2024.12.26.629558v1.80","doi-asserted-by":"publisher","DOI":"10.2202\/1544-6115.1128"},{"key":"2024122716350519000_2024.12.26.629558v1.81","doi-asserted-by":"publisher","DOI":"10.1534\/genetics.118.300864"},{"key":"2024122716350519000_2024.12.26.629558v1.82","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkad393"},{"key":"2024122716350519000_2024.12.26.629558v1.83","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2105-12-35"},{"key":"2024122716350519000_2024.12.26.629558v1.84","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkw377"},{"key":"2024122716350519000_2024.12.26.629558v1.85","doi-asserted-by":"publisher","DOI":"10.1186\/gb-2006-7-10-r100"},{"key":"2024122716350519000_2024.12.26.629558v1.86","doi-asserted-by":"publisher","DOI":"10.3791\/59926"},{"key":"2024122716350519000_2024.12.26.629558v1.87","doi-asserted-by":"publisher","DOI":"10.1016\/j.mce.2015.07.015"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2024.12.26.629558","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T07:45:14Z","timestamp":1768463114000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2024.12.26.629558"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,12,26]]},"references-count":87,"URL":"https:\/\/doi.org\/10.1101\/2024.12.26.629558","relation":{"is-preprint-of":[{"id-type":"doi","id":"10.1016\/j.celrep.2025.115834","asserted-by":"subject"}]},"subject":[],"published":{"date-parts":[[2024,12,26]]},"subtype":"preprint"}}