Kidney Week

Abstract: SA-PO478

ADAM10-MMP14 Complex Regulates Adherens Junction Integrity at Renal Cystogenesis in Autosomal Dominant Polycystic Kidney Disease

Session Information

• Cystic Kidney Diseases: Basic/Translational
  November 09, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Genetic Diseases of the Kidneys

• 1001 Genetic Diseases of the Kidneys: Cystic

Authors

• Xu, Frank, Brigham and Women's Hospital, Boston, Massachusetts, United States

Frank Xu,

• Kong, Tianqing, Brigham and Women's Hospital, Boston, Massachusetts, United States

Tianqing Kong,

• Lu, Tzongshi, Brigham and Women's Hospital, Harvard Medical School, Natick, Massachusetts, United States

Tzongshi Lu,

Background

Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common life-threatening genetic kidney disease and currently do not have relevant therapeutic methods, which eventually results in end-stage renal disease. We previous reported ADPKD is associated with mutations in polycystins, alterations in cell-cell junctions, and disruption of cell polarity in renal epithelial cells. Moreover, our data indicated that Gα12 increase the shedding of E-cadherin, and the inhibition of ADAM10 can block the cystogenesis in renal cells through the preservation of E-Cadherin. In addition, studies also indicate that Matrix metalloproteinase-14 (MMP14) promotes cystogenesis, which was blocked by its inhibitors. However, mechanisms of ADAM10 and MMP14 regulated cystogenesis in ADPKD are not fully understood. In this study, we investigate these two major sheddases association and their roles in renal cystogenesis

Methods

Immunoprecipitation, immunostaining and 3-dimensional (3D) cell culture technologies are used to analyze the interaction between metalloproteinase ADAM10 and MMP14 in Madin-Darby Canine Kidney (MDCK) cells.

Results

Our data shows that ADAM10 and MMP14 associated at hemopexin domain of MMP14 to form a complex, and activation of MMP14 is required for ADAM10 in shedding of E-Cadherin. The enzyme-inactive mutant MMP14^E240A (Glu240 to Ala) or the deletion of the catalytic domain of MMP14 (MMP14CAT) significantly decreases the sheddase activity of ADAM10. In addition, ectopic expression of MMP14 increases the proliferation of MDCK cells, alters the cell-cell adhesion and promotes the cystogenesis of renal epithelial cells in our 3D cell model, in vitro. Our data also shows that knockdown of MMP14 decreases the cleavage of E-Cadherin by ADAM10, and knockdown of ADAM10 enhances the activation of proMMP2 by MMP14. Furthermore, ectopic expression of the MMP14E^240A mutant promotes inhibition of cystogenesis in our 3D epithelial cell models.

Conclusion

Our study shows for the first time that ADAM10 form a unique, stable complex at hemopexin domain of MMP14, and the ADAM10-MMP14 complex play a pivotal role in regulation of their sheddase activity on various cell-cell junctional proteins and cystogenesis in renal epithelial cells. Our data provide a potential therapeutic target in ADPKD through the modulation of ADAM10-MMP14 complex.

Funding

• Private Foundation Support