Mutagenesis of the Cleavage Site of Pro Renin Receptor Abrogates Angiotensin II-Induced Hypertension in Mice

Abstract

It is well demonstrated that activation of renal PRR ([pro]renin receptor) contributes to AngII (angiotensin II)-induced hypertension. Relatively, less is known for the function of sPRR (soluble PRR), the extracellular domain of PRR, primarily generated by S1P (site-1 protease) and furin. Moreover, the relationship between PRR/sPRR and the renin-angiotensin system (RAS) has been debated. In the present study, we used CRISPR/Cas9 strategy to generate mice with mutagenesis of the overlapping cleavage site for both proteases in PRR (termed as PRR R279V/L282V ) to examine the phenotype during AngII infusion with particular emphasis on circulating and intrarenal renin status. PRR R279V/L282V mice exhibited a reduction of sPRR level in plasma by ≈53% and in the kidney by ≈82%, were fertile, and had no gross developmental abnormalities. At basal condition, PRR R279V/L282V mice had drastically suppressed renin levels from plasma, urine, and the kidney as compared to wild-type controls. The hypertensive response of PRR R279V/L282V to AngII infusion was blunted in parallel with attenuated response of intrarenal renin and renal medullary α-epithelial sodium channel expression. By using Ussing chamber technique, primary collecting duct cells from PRR R279V/L282V mice exhibited blunted response of epithelial sodium channel activity to AngII as compared to wild-type cells. Together, these results represent strong evidence favoring sPRR as a mediator of AngII-induced hypertension and a master regulator of renin expression. Therefore, PRR should be considered as an integrative member of the RAS.