{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T15:16:42Z","timestamp":1785511002728,"version":"3.56.0"},"reference-count":32,"publisher":"American Diabetes Association","issue":"1","license":[{"start":{"date-parts":[[2025,10,21]],"date-time":"2025-10-21T00:00:00Z","timestamp":1761004800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.diabetesjournals.org\/content\/license"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["AI166247"],"award-info":[{"award-number":["AI166247"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","award":["DK116073"],"award-info":[{"award-number":["DK116073"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","award":["DK138763"],"award-info":[{"award-number":["DK138763"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["diabetesjournals.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2026,1,1]]},"abstract":"<jats:sec>\n                    <jats:title\/>\n                    <jats:p>Type 1 diabetes (T1D) is characterized by autoimmune destruction of insulin-producing \u03b2-cells. Recent evidence has implicated hybrid insulin peptides (HIPs) as targets of autoreactive CD4 T cells in human T1D patients and as critical autoantigens recognized by diabetogenic T cells in nonobese diabetic (NOD) mice. HIPs form within pancreatic islets through cross-linking reactions between proinsulin fragments and various \u03b2-cell peptides. In the NOD mouse model, highly pathogenic CD4 T cells specifically target HIPs generated through transpeptidation mediated by cathepsin D (CatD). These disease-relevant HIPs consistently incorporate a C-peptide fragment terminating in a critical leucine residue that binds to other \u03b2-cell peptides. In vitro experiments demonstrated that substituting isoleucine for this leucine residue in human C-peptide inhibited CatD-mediated HIP formation. To investigate the in\u00a0vivo significance of this finding, we engineered NOD mice carrying a leucine-to-isoleucine mutation in the insulin 2 gene (NOD INS2I\/I). Mass spectrometric analysis revealed significantly reduced HIP formation in islets from NOD INS2I\/I mice. Significantly decreased activation of HIP-reactive T cells to islets from these mice was also observed. Furthermore, the NOD INS2I\/I mice showed significantly delayed diabetes onset, with 43% remaining disease-free at 1 year compared with only 10% of wild-type NOD controls. These findings implicate HIPs as key mediators in T1D pathogenesis and demonstrate that targeted disruption of HIP formation significantly alters disease progression. Inhibiting CatD-mediated transpeptidation represents a promising therapeutic approach for preventing or delaying T1D onset in genetically susceptible individuals.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Article Highlights<\/jats:title>\n                    <jats:p>Hybrid insulin peptides (HIPs) have been identified as targets of autoreactive T cells in type 1 diabetes, although their causal role in disease pathogenesis has remained unclear. We demonstrated that a single leucine-to-isoleucine substitution in insulin C-peptide significantly disrupts cathepsin D\u2013mediated HIP formation in nonobese diabetic (NOD) mouse islets. NOD mice engineered with this precise modification (NOD INS2I\/I) showed significantly reduced HIP content, decreased T-cell reactivity, and significantly delayed diabetes onset (43% disease-free at 1 year vs. 10% in controls). These findings establish a mechanistic link between HIP formation and disease progression, revealing cathepsin D\u2013mediated transpeptidation as a potential therapeutic target for intervention in at-risk individuals.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.2337\/db25-0324","type":"journal-article","created":{"date-parts":[[2025,10,21]],"date-time":"2025-10-21T20:34:49Z","timestamp":1761078889000},"page":"115-123","update-policy":"https:\/\/doi.org\/10.2337\/ada-journal-policies","source":"Crossref","is-referenced-by-count":6,"title":["Strategic Reduction of Hybrid Insulin Peptide Formation Significantly Delays Diabetes Onset in NOD Mice"],"prefix":"10.2337","volume":"75","author":[{"given":"Jason","family":"Groegler","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical Campus 1 , Aurora, CO"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kaitlin","family":"Mangold","sequence":"additional","affiliation":[{"name":"Department of Immunology and Microbiology, School of Medicine, University of Colorado Anschutz Medical Campus 2 , Aurora, CO"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kelli","family":"Nicholson","sequence":"additional","affiliation":[{"name":"Department of Immunology and Microbiology, School of Medicine, University of Colorado Anschutz Medical Campus 2 , Aurora, CO"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mylinh","family":"Dang","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical Campus 1 , Aurora, CO"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Janet","family":"Wenzlau","sequence":"additional","affiliation":[{"name":"Department of Immunology and Microbiology, School of Medicine, University of Colorado Anschutz Medical Campus 2 , Aurora, CO"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K. Scott","family":"Beard","sequence":"additional","affiliation":[{"name":"Barbara Davis Center for Childhood Diabetes 3 , Aurora, CO"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anita","family":"Hohenstein","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical Campus 1 , Aurora, CO"},{"name":"Department of Immunology and Microbiology, School of Medicine, University of Colorado Anschutz Medical Campus 2 , Aurora, CO"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roger","family":"Powell","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical Campus 1 , Aurora, CO"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8361-8655","authenticated-orcid":false,"given":"Rocky","family":"Baker","sequence":"additional","affiliation":[{"name":"Department of Immunology and Microbiology, School of Medicine, University of Colorado Anschutz Medical Campus 2 , Aurora, CO"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5430-6254","authenticated-orcid":false,"given":"Kathryn","family":"Haskins","sequence":"additional","affiliation":[{"name":"Department of Immunology and Microbiology, School of Medicine, University of Colorado Anschutz Medical Campus 2 , Aurora, CO"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer","family":"Matsuda","sequence":"additional","affiliation":[{"name":"Department of Immunology and Microbiology, School of Medicine, University of Colorado Anschutz Medical Campus 2 , Aurora, 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