Functional characterization of two novel germline mutations of the KCNJ5 gene in hypertensive patients without primary aldosteronism but with ACTH-dependent aldosterone hypersecretion

Clin Endocrinol (Oxf). 2016 Dec;85(6):845-851. doi: 10.1111/cen.13132. Epub 2016 Jul 12.

Abstract

Background: Germline mutations of the KCNJ5 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 (APAs) and sporadic cases of primary aldosteronism (PA).

Objective: To present two novel KCNJ5 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 CYP11B1/CYP11B2 chimeric gene and KCNJ5 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 KCNJ5 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 CYP11B1/CYP11B2 chimeric gene was not detected.

Conclusions: These findings expand on the clinical spectrum of phenotypes associated with KCNJ5 mutations and implicate these mutations in the pathogenesis of hypertension associated with increased aldosterone response to ACTH stimulation.

Publication types

  • Case Reports

MeSH terms

  • Adrenocorticotropic Hormone / pharmacology*
  • Aldosterone / metabolism*
  • Cytochrome P-450 CYP11B2 / genetics
  • Electrophysiological Phenomena
  • Female
  • G Protein-Coupled Inwardly-Rectifying Potassium Channels / genetics*
  • Genetic Association Studies
  • Germ-Line Mutation / physiology*
  • Humans
  • Hyperaldosteronism
  • Hypertension / etiology*
  • Male
  • Middle Aged
  • Steroid 11-beta-Hydroxylase / genetics

Substances

  • G Protein-Coupled Inwardly-Rectifying Potassium Channels
  • KCNJ5 protein, human
  • Aldosterone
  • Adrenocorticotropic Hormone
  • Cytochrome P-450 CYP11B2
  • Steroid 11-beta-Hydroxylase