Transmembrane Serine Protease 2 and Proteolytic Activation of the Epithelial Sodium Channel in Mouse Kidney

Key Points Proteolytic activation of the epithelial sodium channel (ENaC) was compromised by transmembrane serine protease 2 deficiency in murine cortical collecting duct cells and native mouse kidney. To compensate for impaired ENaC activation, rise in plasma aldosterone in response to low-salt diet was enhanced in Tmprss2 −/− mice. Transmembrane serine protease 2 may be a potential drug target to limit proteolytic ENaC activation in disorders with increased renal ENaC activity. Background The renal epithelial sodium channel (ENaC) is essential for sodium balance and BP control. ENaC undergoes complex proteolytic activation by not yet clearly identified tubular proteases. Here, we examined a potential role of transmembrane serine protease 2 (TMPRSS2). Methods Murine ENaC and TMPRSS2 were (co)expressed in Xenopus laevis oocytes. ENaC cleavage and function were studied in TMPRSS2-deficient murine cortical collecting duct (mCCD cl1 ) cells and TMPRSS2-knockout ( Tmprss2 −/− ) mice. Short-circuit currents ( I SC ) were measured to assess ENaC-mediated transepithelial sodium transport of mCCD cl1 cells. The mCCD cl1 cell transcriptome was studied using RNA sequencing. The effect of low-sodium diet with or without high potassium were compared in Tmprss2 −/− and wild-type mice using metabolic cages. ENaC-mediated whole-cell currents were recorded from microdissected tubules of Tmprss2 −/− and wild-type mice. Results In oocytes, coexpression of murine TMPRSS2 and ENaC resulted in fully cleaved γ -ENaC and approximately two-fold stimulation of ENaC currents. High baseline expression of TMPRSS2 was detected in mCCD cl1 cells without a stimulatory effect of aldosterone on its function or transcription. TMPRSS2 knockout in mCCD cl1 cells compromised γ -ENaC cleavage and reduced baseline and aldosterone-stimulated I SC , which could be rescued by chymotrypsin. A compensatory transcriptional upregulation of other proteases was not observed. Tmprss2 −/− mice kept on standard diet exhibited no apparent phenotype, but renal γ -ENaC cleavage was altered. In response to a low-salt diet, particularly with high potassium intake, Tmprss2 −/− mice increased plasma aldosterone significantly more than wild-type mice to achieve a similar reduction of renal sodium excretion. Importantly, the stimulatory effect of trypsin on renal tubular ENaC currents was much more pronounced in Tmprss2 −/− mice than that in wild-type mice. This indicated the presence of incompletely cleaved and less active channels at the cell surface of TMPRSS2-deficient tubular epithelial cells. Conclusions TMPRSS2 contributes to proteolytic ENaC activation in mouse kidney in vivo .