Functional characterization of two novel germline mutations of the <i><scp>KCNJ</scp>5</i> gene in hypertensive patients without primary aldosteronism but with <scp>ACTH</scp>‐dependent aldosterone hypersecretion

Summary Background Germline mutations of the KCNJ 5 gene encoding Kir3·4, a member of the inwardly rectifying K + channel, have been identified in ‘normal’ adrenal glands, patients with familial hyperaldosteronism ( FH ) type III , aldosterone‐producing adenomas ( APA s) and sporadic cases of primary aldosteronism ( PA ). Objective To present two novel KCNJ 5 gene mutations in hypertensive patients without PA , but with Adrenocorticotropic hormone (ACTH)‐dependent aldosterone hypersecretion. Design and Patients Two hypertensive patients without PA , who exhibited enhanced ACTH ‐dependent response of aldosterone secretion, underwent genetic testing for the presence of the CYP 11B1/ CYP 11B2 chimeric gene and KCNJ 5 gene mutations. Genomic DNA was isolated from peripheral white blood cells, and the exons of the entire coding regions of the above genes were amplified and sequenced. Electrophysiological studies were performed to determine the effect of identified mutation(s) on the membrane reversal potentials. Structural biology studies were also carried out. Results Two novel germline heterozygous KCNJ 5 mutations, p.V259M and p.Y348N, were detected in the two subjects. Electrophysiological studies showed that the Y348N mutation resulted in significantly less negative reversal potentials, suggesting loss of ion selectivity, while the V259M mutation did not affect the Kir3.4 current. In the mutated structural biology model, the N348 mutant resulted in significant loss of the ability for hydrogen bonding, while the M259 mutant was capable of establishing weaker interactions. The CYP 11B1/ CYP 11B2 chimeric gene was not detected. Conclusions These findings expand on the clinical spectrum of phenotypes associated with KCNJ 5 mutations and implicate these mutations in the pathogenesis of hypertension associated with increased aldosterone response to ACTH stimulation.