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Introduction: Activation of lung mesenchymal cells is key to scleroderma-associated interstitial lung disease (SSc-ILD) and idiopathic pulmonary fibrosis (IPF). Both epigenetic alterations and activation of pro-fibrotic signaling pathways, among which Transforming growth factor-beta1 (TGFB1) is prominent, contribute to mesenchymal cell activation in these diseases. Methods: Proteomics profiling of cell fractions was performed in lung mesenchymal cells from Controls without lung disease (n=5) and from subjects with IPF (n=5) or SSc-ILD (n=3). Candidate phenotype regulators were identified based on their expression level, common deregulation between fibrotic cell groups, and literature analysis. The pathophysiological relevance of 24 candidate hits was assessed in RNA interference assays in a 48-well format, using pro-collagen-1 (COL1) ELISA as a readout. Results: Functional annotation classes most upregulated in IPF and SSc-ILD cells were Actin Cytoskeleton Organization and Cell Adhesion , consistent with alterations in cell-matrix interactions, as well as Protein Localization . Protein Transport and Membrane-Bounded Vesicle were most downregulated, consistent with alterations in the endocytic regulation of signaling pathways. Of the selected candidates, 8 (SIRPA, FRYL, DLST, FHL2, MAOA, PRDX3, RAB31, LTBP2) controlled COL1 expression. Conclusion: Subcellular proteomics followed with medium-throughput RNA interference assays identified 8 novel molecular determinants of COL1 expression in lung mesenchymal cells. Funded by grants from PRES Sorbonne Paris Cite, GFRS, and Intermune. |
