Contribution of Extracellular Matrix and Signal Mechanotransduction to Epithelial Cell Damage in Inflammatory Bowel Disease Patients: A Proteomic Study

Abstract

This study utilizes 2D-DIGE (difference gel etrophoresis), isotope-coded protein labeling and biochemical assays to characterize protein alteration in ulcerative colitis (UC) and Crohn's disease (CD) in human epithelial cell and mucosal biopsies in inflammatory bowel disease (IBD)-affected patients. The aim of this study is to identify the key molecular signatures involved in epithelial cell structure of IBDs. In non-inflamed UC (QUC) keratins, vimentin, and focal adhesion kinase (7) increased, whereas vinculin and de-tyrosinated α-tubulin decreased; inflammation (IUC) exacerbated molecular changes, being collagen type VI alpha 1 chain (COL6A1), tenascin-C and vimentin increased. In non-inflamed CD (QCD), tenascin C, de-tyrosinated α-tubulin, vinculin, FAK, and Rho-associated protein kinase 1 (ROCK1) decreased while vimentin increased. In inflamed CD (ICD), COL6A1, vimentin and integrin alpha 4 increased. In QUC, cell metabolism is characterized by a decrease of the tricarboxylic acid cycle enzymes and a decrease of short/branched chain specific acyl-CoA dehydrogenase, fatty acid synthase, proliferator-activated receptors alpha, and proliferator-activated receptors gamma. In QCD a metabolic rewiring occurs, as suggested by glycerol-3-phosphate dehydrogenase (GPD2), pyruvate dehydrogenase E1 component subunit beta, NADH dehydrogenase [ubiquinone] iron-sulfur protein 3, and 4-trimethylaminobutyraldehyde dehydrogenase increment, while dihydrolipoyl dehydrogenase decreased. Macroautophagy is activated in QUC and IUC, with increased levels of p62, HSC70, major vault protein, myosin heavy chain 9, whereas it is blunted in QCD and ICD. The differing pattern of extracellular matrix, cytoskeletal derangements, cellular metabolism, and autophagy in UC and CD may contribute to the pathophysiological understanding of these disorders and serve as diagnostic markers in IBD patients.

Protein Deimination and Extracellular Vesicle Profiles in Antarctic Seabirds

“…Collagen alpha-4 (VI) chain was found deiminated in wandering albatross plasma. It is an extracellular matrix protein [206], is found in lymphoid tissues [207] and has roles, amongst others, in cellular and mucosal immunity and inflammatory diseases such as ulcerative colitis and membranous glomerulonephritis [208,209]. In birds, it has roles in CNS development relating to plasticity and axon growth in the chicken [210].…”

ECM Remodeling in Breast Cancer with Different Grade: Contribution of 2D‐DIGE Proteomics

“…Our results indicate that MSN, entailed in connections of major cytoskeletal structures to the plasma membrane, A1BG, and two proteoforms of FGB increased, suggesting that a matrix remodeling characterized DN‐E3G3 . Previous studies from our group in inflammatory bowel disease (IBD) patients, in COL6A1 deficient animal models and mypoatic patients with mutation in COL6A1, highlighted the role of matrix, metabolism, and structural remodeling in disease onset and progression. A similar “mechanosignaling” appears active also in DN‐E3G3 suggesting that matrix remodeling can be able to perceive physical and chemical signals activating focal adhesion that sense the rigidity of the extracellular matrix .…”

“…The 2D‐DIGE is a top‐down proteomic technique for the study of a fraction of metabolic, structural, cytoskeletal, and cell signaling protein levels and provides a picture of proteoforms and PTMs of undigested proteins . Considering E3G3 characteristics according to tumor hormone receptors expression, ECM proteins, in particular COL6A1, were more abundant in DN‐E3G3 than in E3G3 BCs.…”

“…Multivariate analysis of distant metastasis‐free survival in untreated breast tumor patients revealed a significant interaction between ECM3 and histological grade, indicating that ECM3 was significantly associated with high risk of relapse in patients with undifferentiated (grade III) tumors. Recent independent studies by our group have highlighted the function of ECM in different disorders introducing a new view of ECM as active player, pinpointing its role as signal “mechanotransducer” from outside to inside cells influencing structural organization, metabolism, and nuclear signaling . Furthermore, it has been found that tissue mechanics, assisted by ECM remodeling, is a key regulator in cancer progression in which structural forces and biochemical signals modulate cell and tissue behavior eliciting a specific cellular response …”

A Fragment of Collagen Type VI alpha-3 chain is Elevated in Serum from Patients with Gastrointestinal Disorders

“…In IBD COL6α1, COL6α2 and COL6α3 chains are elevated in the intestinal tissue in patients with CD and UC, and COL6 has been demonstrated to be involved in the integrity of the epithelial barrier 20,21 . It has previously been demonstrated that COL6 turnover was elevated in luminal CD and stricturing CD compared to fistulizing CD 22 .…”