Rationale: The lung extracellular matrix (ECM) in idiopathic pulmonary fibrosis (IPF) mediates progression of fibrosis by decreasing fibroblast expression of miR-29 (microRNA-29), a master negative regulator of ECM production. The molecular mechanism is undefined. IPF-ECM is stiffer than normal. Stiffness drives fibroblast ECM production in a YAP (yes-associated protein)-dependent manner, and YAP is a known regulator of miR-29. Therefore,we tested the hypothesis that negative regulation of miR-29 by IPF-ECM was mediated by mechanotransduction of stiffness. Objectives: To determine how IPF-ECM negatively regulates miR-29. Methods: We decellularized lung ECM using detergents and prepared polyacrylamide hydrogels of defined stiffness by varying acrylamide concentrations. Mechanistic studies were guided by immunohistochemistry of IPF lung and used cell culture, RNAbinding protein assays, and xenograft models. Measurements and Main Results: Contrary to our hypothesis, we excluded fibroblast mechanotransduction of ECM stiffness as the primary mechanism deregulating miR-29. Instead, systematic examination of miR-29 biogenesis revealed a microRNA processing defect that impeded processing ofmiR-29 into its mature bioactive forms. Immunohistochemical analysis of the microRNA processing machinery in IPF lung specimens revealed decreased Dicer1 expression in the procollagen-rich myofibroblastic core of fibroblastic foci compared with the focus perimeter and adjacent alveolar walls. Mechanistically, IPF-ECM increased association of the Dicer1 transcript with RNA binding protein AUF1 (AU-binding factor 1), and Dicer1 knockdown conferred primary human lung fibroblasts with cell-autonomous fibrogenicity in zebrafish and mouse lung xenograft models. Conclusions: Our data identify suppression of fibroblast Dicer1 expression in the myofibroblast-rich IPF fibroblastic focus core as a central step in the mechanism by which the ECM sustains fibrosis progression in IPF.
|Original language||English (US)|
|Number of pages||11|
|Journal||American journal of respiratory and critical care medicine|
|State||Published - Aug 15 2018|
Bibliographical noteFunding Information:
Supported by the NIH grants R01 HL125236 (P.B.B.), P01 HL091775 and R01 HL125227 (C.A.H.), and T32 HL077410 (J.H. and D.J.B.); National Center for Advancing Translational Sciences grant UL1TR000114 (C.F.); National Science Foundation Graduate Research Fellowship Program grant 00039202 (C.E.K.); and funds provided by the O’Brien and Witowski families. C.E.K. is a recipient of the Richard Pyle Scholar Award from the Achievement Rewards for College Scientists Foundation.
- Extracellular matrix
- Idiopathic pulmonary fibrosis
- Yes-associated protein