Identification of key pro-survival proteins in isolated colonic goblet cells of Winnie, a murine model of spontaneous colitis
Wilson, RR and Gundamaraju, R and Vemuri, R and Angelucci, C and Geraghty, DP and Guven, N and Eri, RD, Identification of key pro-survival proteins in isolated colonic goblet cells of Winnie, a murine model of spontaneous colitis, Inflammatory Bowel Diseases pp. 1-13. ISSN 1536-4844 (2019) [Refereed Article]
Background: Accumulating evidence suggests that the goblet cell-derived mucin-2 (Muc2) is a major component of the immune system and that perturbations in Muc2 lead to an ulcerative colitis-like phenotype. The animal model Winnie carries a missense mutation in Muc2 that causes Muc2 misfolding, accumulation in goblet cells and ER stress. Excessive ER stress is a hallmark of many diseases, including ulcerative colitis, cancer, diabetes and Parkinsonís disease. However, rather than committing to cell death, the typical outcome of unresolved ER stress, Winnie goblet cells are characterised by hyper proliferation, suggesting additional regulation of this cellular stress response.
Methods: To elucidate the molecular mechanisms underlying ulcerative colitis in the Winnie model we isolated goblet cells from Winnie and wild type mice and used label-free quantitative proteomics and bioinformatics to understand the functional consequences of Muc2 misfolding and accumulation.
Results: A large number of changes were identified that highlight a dramatic reprogramming of energy production, including enhanced utilization of butyrate, a key energy source of colonic cells. A major finding was the marked up-regulation of the coiled-coil-helix-coiled-coil-helix domain proteins Chchd2, Chchd3 and Chchd6. In particular, we identified and confirmed the upregulation and nuclear translocation of Chchd2, a protein known to inhibit oxidative stress induced apoptosis.
Conclusions: This study is the first to apply proteome-level analysis to the pre-clinical Winnie model of ulcerative colitis. Identification of proteins and pathways affected in isolated Winnie goblet cells provides evidence for novel adaptive mechanisms underlying cell survival under conditions of chronic ER stress.
ER stress, ulcerative colitis, proteomics, apoptosis