I-mfa, Mesangial Cell TRPC1 Channel, and Regulation of Glomerular Filtration Rate

Background: Inhibitor of MyoD family A (I-mfa) is a cytosolic protein. Its function in kidney is unknown. The aim of the present study was to examine the regulatory role of I-mfa on glomerular filtration rate (GFR). Methods: GFR was measured by transdermal measurement of FITC-sinitrin clearance in conscious wild type (WT) and I-mfa knockout (KO) mice. Cell contractility was assessed in a single human or mouse mesangial cell. Single cell RNA sequence (scRNA-seq), Western blot, and Ca 2+ imaging were used to evaluate the effects of I-mfa on TRPCs at messenger, protein and functional levels in MCs. Results: In KO mice, GFR was significantly lower than that in WT mice. In WT mice, knocking down I-mfa selectively in mesangial cells using targeted nanoparticle/siRNA delivery system significantly decreased GFR. In human mesangial cells, overexpression of I-mfa significantly blunted the Ang II-stimulated contraction, and knockdown of I-mfa significantly enhanced the contractile response. Consistently, the Ang II-induced contraction was significantly augmented in primary mesangial cells isolated from KO mice. The exaggerated response was restored by re-introducing I-mfa. Furthermore, scRNA-seq showed an increase in trpc1 messenger and Western blot showed an increase in TRPC1 protein abundance in I-mfa KO mouse mesangial cells. TRPC1 protein abundance was decreased in HEK cells overexpressing I-mfa. Ca 2+ imaging experiments showed that downregulation of I-mfa significantly enhanced Ang II-stimulated Ca 2+ entry in human mesangial cells. Finally, TRPC1 inhibitor, Pico145 significantly blunted Ang II-induced mesangial cell contraction. Conclusions: I-mfa positively regulated GFR by decreasing mesangial cell contractile function through inhibition of TRPC1-mediated Ca 2+ signaling.