MLK3 mediates impact of PKG1α on cardiac function and controls blood pressure through separate mechanisms

JCI Insight. 2021 Sep 22;6(18):e149075. doi: 10.1172/jci.insight.149075.

Abstract

cGMP-dependent protein kinase 1α (PKG1α) promotes left ventricle (LV) compensation after pressure overload. PKG1-activating drugs improve heart failure (HF) outcomes but are limited by vasodilation-induced hypotension. Signaling molecules that mediate PKG1α cardiac therapeutic effects but do not promote PKG1α-induced hypotension could therefore represent improved therapeutic targets. We investigated roles of mixed lineage kinase 3 (MLK3) in mediating PKG1α effects on LV function after pressure overload and in regulating BP. In a transaortic constriction HF model, PKG activation with sildenafil preserved LV function in MLK3+/+ but not MLK3-/- littermates. MLK3 coimmunoprecipitated with PKG1α. MLK3-PKG1α cointeraction decreased in failing LVs. PKG1α phosphorylated MLK3 on Thr277/Ser281 sites required for kinase activation. MLK3-/- mice displayed hypertension and increased arterial stiffness, though PKG stimulation with sildenafil or the soluble guanylate cyclase (sGC) stimulator BAY41-2272 still reduced BP in MLK3-/- mice. MLK3 kinase inhibition with URMC-099 did not affect BP but induced LV dysfunction in mice. These data reveal MLK3 as a PKG1α substrate mediating PKG1α preservation of LV function but not acute PKG1α BP effects. Mechanistically, MLK3 kinase-dependent effects preserved LV function, whereas MLK3 kinase-independent signaling regulated BP. These findings suggest augmenting MLK3 kinase activity could preserve LV function in HF but avoid hypotension from PKG1α activation.

Keywords: Cardiology; Cyclic nucleotides; Heart failure; Hypertension.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Aorta / pathology
  • Blood Pressure / drug effects
  • Blood Pressure / genetics
  • Cyclic GMP-Dependent Protein Kinase Type I / metabolism*
  • HEK293 Cells
  • Heart Failure / complications
  • Heart Failure / physiopathology*
  • Humans
  • Hypertension / genetics
  • MAP Kinase Kinase Kinases / antagonists & inhibitors
  • MAP Kinase Kinase Kinases / genetics*
  • MAP Kinase Kinase Kinases / metabolism*
  • Male
  • Mice
  • Mice, Knockout
  • Mitogen-Activated Protein Kinase Kinase Kinase 11
  • Muscle, Smooth, Vascular / pathology
  • Myocytes, Smooth Muscle / pathology
  • Phosphorylation
  • Protein Kinase Inhibitors / pharmacology
  • Pyrazoles / pharmacology
  • Pyridines / pharmacology
  • Pyrroles / pharmacology
  • Sildenafil Citrate / pharmacology
  • Vascular Stiffness / genetics
  • Vasodilator Agents / pharmacology
  • Ventricular Dysfunction, Left / etiology
  • Ventricular Dysfunction, Left / physiopathology*

Substances

  • 3-(4-Amino-5-cyclopropylpyrimidine-2-yl)-1-(2-fluorobenzyl)-1H-pyrazolo(3,4-b)pyridine
  • Protein Kinase Inhibitors
  • Pyrazoles
  • Pyridines
  • Pyrroles
  • URMC-099
  • Vasodilator Agents
  • Sildenafil Citrate
  • Cyclic GMP-Dependent Protein Kinase Type I
  • MAP Kinase Kinase Kinases