Dysregulation of metalloproteinase complex ADAM10‐MMP14 plays the key role in preservation of adherens junction integrity at renal cystogenesis

Abstract

BackgroundAutosomal dominant polycystic kidney disease (ADPKD) is one of the most common lifeth‐reatening genetic diseases and is characterized by early cystogenesis in kidney, which eventually results in end‐stage renal disease. We previously reported that the inhibition of A Disintegrin and metalloproteinase domain‐containing protein 10 (ADAM10) can block the cystogenesis in renal tubular epithelial cells through the preservation of E‐Cadherin. In addition, recent studies also indicate that another major matrix metalloproteinase, Matrix metalloproteinase‐14 (MMP14) promotes cystogenesis in kidney epithelial cells, which was blocked by its inhibitors. In our study, we investigate these two major sheddases association and their roles in preservation of adherens junctions in renal cystogenesisMethodsImmunoprecipitation, immunostaining and 3‐D cell culture technologies are used to analyze the interaction between metalloproteinase ADAM10 and MMP14 in Madin‐Darby Canine Kidney (MDCK) cells.ResultsOur data shows that ADAM10 and MMP14 form a stable complex on the membrane of MDCK cells at hemopexin domain of MMP14. The activation of MMP14 is required for ADAM10 in shedding of E‐Cadherin. Either an enzyme‐inactive mutant MMP14E240A (Glu240 to Ala) or the deletion of the catalytic domain of MMP14 (MMP14ΔCAT) significantly decreases the sheddase activity of ADAM10. In addition, knockdown of MMP14 decreases the cleavage of ECadherin by ADAM10, and knockdown of ADAM10 enhances the activation of proMMP2 by MMP14. Furthermore, ectopic expression of MMP14 increases the proliferation of renal epithelial cells, alters the cell‐cell adhesion and promotes the cystogenesis of renal epithelial cells in 3‐dimensional collagen‐1 gels. Ectopic expression of the MMP14E240A mutant promotes strong contact inhibition of the cells, and results in tubular rather than cystic growth in collagen gels.ConclusionOur study shows for the first time that ADAM10 form a unique, stable complex at hemopexin domain of MMP14 on the surface of epithelial cells, and the ADAM10‐MMP14 complex regulates their sheddase activity on many cell junctional proteins. Furthermore, ADAM10 is a negative regulator of MMP14 that ADAM10 inhibits the shaddase activity of MMP14. The functional interaction between ADAM10 and MMP14 as a complex could play a pivotal role in the cystogenesis of renal epithelial cells. Our data provide a potential therapeutic target in ADPKD through the modulation of ADAM10‐MMP14 complex.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.