sodium-dodecyl-sulfate and Fibrosis

Chronic inflammation is often associated with the development of tissue fibrosis, but how mesenchymal cell responses dictate pathological fibrosis versus resolution and healing remains unclear. Defining stromal heterogeneity and identifying molecular circuits driving extracellular matrix deposition and remodeling stands to illuminate the relationship between inflammation, fibrosis, and healing. We performed single-cell RNA-sequencing of colon-derived stromal cells and identified distinct classes of fibroblasts with gene signatures that are differentially regulated by chronic inflammation, including IL-11-producing inflammatory fibroblasts. We further identify a transcriptional program associated with trans-differentiation of mucosa-associated fibroblasts and define a functional gene signature associated with matrix deposition and remodeling in the inflamed colon. Our analysis supports a critical role for the metalloprotease Adamdec1 at the interface between tissue remodeling and healing during colitis, demonstrating its requirement for colon epithelial integrity. These findings provide mechanistic insight into how inflammation perturbs stromal cell behaviors to drive fibroblastic responses controlling mucosal matrix remodeling and healing.

Topics: ADAM Proteins; Animals; Cell Differentiation; Colitis; Colon; Extracellular Matrix; Fibroblasts; Fibrosis; Gene Expression Regulation; Humans; Inflammation; Interleukin-11; Intestinal Mucosa; Male; Mesenchymal Stem Cells; Mice; Mice, Inbred C57BL; Sequence Analysis, RNA; Single-Cell Analysis; Sodium Dodecyl Sulfate; Transcription, Genetic; Transcriptome; Wound Healing

Cardiac fibrosis is characterized by excessive deposition of extracellular matrix (ECM) and is a common complication of various cardiovascular diseases. However, little is known about proteins in the cardiac extracellular space. Proteomics analysis of cardiac ECM can be challenging due to the presence of more abundant intracellular proteins, the low degree of solubility of integral ECM proteins, and the presence of abundant posttranslational modifications. Here we describe an extraction methodology based on tissue decellularization, which allows the biochemical subfractionation of extracellular proteins in cardiac tissue. These relatively low-complexity protein fractions are suitable for analysis by gel-LC-MS/MS and other proteomics techniques.

Topics: Animals; Chromatography, Liquid; Extracellular Matrix; Extracellular Matrix Proteins; Fibrosis; Guanidine; Humans; Liquid-Liquid Extraction; Myocardial Reperfusion Injury; Myocardium; Peptide Fragments; Peptide Mapping; Proteolysis; Proteomics; Sodium Chloride; Sodium Dodecyl Sulfate; Swine; Tandem Mass Spectrometry