Inhibition of Anoctamin 3 Attenuates Cystogenesis in Autosomal Dominant Polycystic Kidney Disease

Background: Autosomal dominant polycystic kidney disease(ADPKD) is the most common inherited kidney disease,which is characterized by progressive growth of renal cysts.As we reported Anoctamin 1,one of chlorine channels family promotes both ciliogenesis and cyst formation in PKDcells.Wherever another member Anoctamin3(ANO3) was recently shown to be involved in ADPKD progression in our research.It seems there is nobody reported. Methods: 1.RT RCR,WB and Immunohistochemistry to detect ANO3 in kidney tissue of ADPKD patients and normal kidndy tissue. 2.3Dculture to vertify the function of ANO3 in cysts formation in RCTE and IMCD3 cell with siANO3. 3.Co-immunofluorescence to investigate the impact of ANO3 on cilia and PC2 by siANO3. 4.WB to test the effect of ANO3 in proliferation signaling pathways. 5.To measure the kidney cyst index of Ano3-/-:Pkd1 flox/flox:CAG-creER+mice by HE staining and MRI. Results: 1.ANO3 located in renal tubules and renal cyst and high expression in human and mouse ADPKD kidneys. 2.shANO3 attenuated cyst formation in 3D culture cells. 3.ANO3 was localization on ciliary body and shANO3 increased the cilia length and the expression of PC2. 4.ShANO3 could attenuated cyst formation by inhibiting phosphorylation of AKT and mTOR signal pathways. 5.Cyst index of PKD1flox/floxCAG-creER+mice was significantly decrease than which in PKD1RC/RCmice. Conclusion: Our data indicate inhibition of Anoctamin3 attenuates cystogenesis in ADPKD by decreasing phosphorylation of AKT and mTOR signal pathways.