Molecular Mechanism for Hypertensive Renal Disease: Differential Regulation of Chromogranin A Expression at 3'-Untranslated Region Polymorphism C+87T by MicroRNA-107

J Am Soc Nephrol. 2015 Aug;26(8):1816-25. doi: 10.1681/ASN.2014060537. Epub 2014 Nov 12.

Abstract

Chromogranin A (CHGA) is coreleased with catecholamines from secretory vesicles in adrenal medulla and sympathetic axons. Genetic variation in the CHGA 3'-region has been associated with autonomic control of circulation, hypertension, and hypertensive nephropathy, and the CHGA 3'-untranslated region (3'-UTR) variant C+87T (rs7610) displayed peak associations with these traits in humans. Here, we explored the molecular mechanisms underlying these associations. C+87T occurred in a microRNA-107 (miR-107) motif (match: T>C), and CHGA mRNA expression varied inversely with miR-107 abundance. In cells transfected with chimeric luciferase/CHGA 3'-UTR reporters encoding either the T allele or the C allele, changes in miR-107 expression levels had much greater effects on expression of the T allele. Cotransfection experiments with hsa-miR-107 oligonucleotides and eukaryotic CHGA plasmids produced similar results. Notably, an in vitro CHGA transcription/translation experiment revealed that changes in hsa-miR-107 expression altered expression of the T allele variant only. Mice with targeted ablation of Chga exhibited greater eGFR. Using BAC transgenesis, we created a mouse model with a humanized CHGA locus (T/T genotype at C+87T), in which treatment with a hsa-miR-107 inhibitor yielded prolonged falls in SBP/DBP compared with wild-type mice. We conclude that the CHGA 3'-UTR C+87T disrupts an miR-107 motif, with differential effects on CHGA expression, and that a cis:trans (mRNA:miR) interaction regulates the association of CHGA with BP and hypertensive nephropathy. These results indicate new strategies for probing autonomic circulatory control and ultimately, susceptibility to hypertensive renal sequelae.

Keywords: CKD; hypertension; molecular genetics; progression of renal failure; systolic BP; transgenic mouse.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • 3' Untranslated Regions
  • Alleles
  • Animals
  • Blood Pressure
  • Chromogranin A / genetics*
  • Chromogranin A / metabolism
  • Glomerular Filtration Rate
  • HEK293 Cells
  • Humans
  • Hypertension, Renal / genetics*
  • Hypertension, Renal / metabolism
  • Luciferases
  • Male
  • Mice
  • Mice, Transgenic
  • MicroRNAs / genetics*
  • PC12 Cells
  • Polymorphism, Genetic
  • Rats

Substances

  • 3' Untranslated Regions
  • CHGA protein, human
  • Chromogranin A
  • MIRN107 microRNA, human
  • MicroRNAs
  • Luciferases