Mutation of KCNJ5 induced adrenal aldosterone adenoma via CYP11A1 regulation

Primary aldosteronism (PA), also known as Conn’s syndrome or adrenal aldosterone producing adenoma (APA), is predominantly caused by functional adrenal tumors and is a common cause of secondary hypertension. The KCNJ5 gene mutations are frequently associated with APA, leading to increased aldosterone production. This study investigates the effect of KCNJ5 mutations on CYP11A1 and the aldosterone biosynthesis pathway. We created two model cell lines by introducing homozygous p.L168R and p.G151R mutations in the KCNJ5 gene to SW13 cell line using CRISPR/Cas9 technology. The mutations were verified succeed through Sanger sequencing and multi-omics analysis. Aldosterone and its biosynthesis intermediates were quantitatively analyzed, and the expression of CYP11A1 mRNA in wild-type and mutant cell lines was detected through qPCR. Increased concentrations of aldosterone, pregnenolone, progesterone, and corticosterone in the KCNJ5 mutated cell lines were observed compared to the wild type, while cholesterol levels remained unchanged. qPCR results showed increased CYP11A1 mRNA expression in KCNJ5 mutant cells. Therefore, KCNJ5 mutations promote aldosterone synthesis by enhancing CYP11A1 activity, which catalyzes the conversion of cholesterol to pregnenolone, a critical step in the aldosterone biosynthesis pathway. This study highlights the potential of CYP11A1 as a therapeutic target for treating APA-induced secondary hypertension.