Regulation of cardiac ion channel function via allosteric modulators

Summary

Principal Investigator: ANDREW ROBERT MARKS
Abstract: The ryanodine receptor (RyR) is comprised of 4 RyR protomers and proteins that bind to the cytoplasmic domain of the channel forming a macromolecular signaling complex. This proposal addresses the mechanisms by which allosteric modulators regulate RyR function. Two specific forms of allosteric modulation will be examined: 1) regulation of the channel by derivatives of 1,4-benzothiazepines that potently effect channel gating via allosteric effects; 2) regulation of the channel by protein-protein interactions with the stabilizing subunit FKBP12/12.6 (calstabin1/2). Four aims are proposed: Aim 1: Allosteric regulation of RyR2 by small molecules that enhance binding of calstabin2 to RyR2. Effects of 1,4- benzothiazepine derivatives on RyR2 channel function will be examined using RyR2 channels reconstituted into planar lipid bilayers. The binding site on RyR2 for the 1,4-benzothiazepine derivatives will be identified using photoaffinity radiolabels. The hypothesis is derivatives of 1,4-benzothiazepines bind to and allosterically modulate the function of RyR2 and RyR1. Aim 2: Allosteric modulation of RyR2 as a mechanism for preventing cardiac arrhythmias RyR2 are PKA hyperphosphorylated and "leaky" in atrial fibrillation (AF) and JTV519 prevents exercised induced cardiac arrhythmias in WT and calstabin2+/- mice but not in calstabin2-/- mice indicating that the mechanism of action of this novel anti-arrhythmic drug requires calstabin2. The hypothesis is small molecules that enhance calstabin2 binding to RyR2 can prevent cardiac arrhythmias via allosteric modulation of RyR2. Using genetic mouse models harboring RyR2 mutations linked to sudden cardiac death in humans, and RyR2 mutations that mimic constitutively PKA phosphorylated or non-phosphorylatable RyR2 and animal models of AF, and myocardial infarction, we will determine whether enhancing binding of calstabin2 to RyR2 prevents cardiac arrhythmias. Aim 3: Stabilization of calstabin2 binding to RyR2 as a mechanism for treating heart failure (HF). RyR2 are PKA hyperphosphorylated and depleted of calstabin2 in HF and JTV519 improves cardiac function in WT and calstabin2+/- mice but not in calstabin2-/- mice. Using genetic mouse models and models of myocardial infarction, we will determine whether enhancing binding of calstabin2 to RyR2 using JTV519 that modify RyR2 via allosteric effects improve cardiac function in HF. Skeletal muscle fatigue is increased and RyR1 are PKA hyperphosphorylated and depleted of calstabin1 in HF, and JTV519 induces rebinding of calstabin1 to RyR1 probably via an allosteric effect on the channel. Using animal models of myocardial infarction and HF, we will investigate whether JTV519 improves skeletal muscle function in HF. The studies are significant because they may lead to a novel therapeutic approach based on allosteric modulation of RyR that can result in improved therapy for human cardiovascular diseases.
Funding Period: 2005-12-20 - 2007-11-30
more information: NIH RePORT

Top Publications

  1. pmc Ryanodine receptor studies using genetically engineered mice
    Alexander Kushnir
    Department of Physiology and Cellular Biophysics, Clyde and Helen Wu Center for Molecular Cardiology, New York, NY, USA
    FEBS Lett 584:1956-65. 2010
  2. pmc Dysfunctional ryanodine receptors in the heart: new insights into complex cardiovascular diseases
    Steven O Marx
    Division of Cardiology, College of Physicians and Surgeons of Columbia University, New York, NY 10032, USA
    J Mol Cell Cardiol 58:225-31. 2013
  3. pmc Ryanodine receptor channelopathies
    Matthew J Betzenhauser
    Department of Physiology, Clyde and Helen Wu Center for Molecular Cardiology, Columbia University College of Physicians and Surgeons, 630 West 168th Street, New York, NY 10032, USA
    Pflugers Arch 460:467-80. 2010

Detail Information

Publications4

  1. pmc Ryanodine receptor studies using genetically engineered mice
    Alexander Kushnir
    Department of Physiology and Cellular Biophysics, Clyde and Helen Wu Center for Molecular Cardiology, New York, NY, USA
    FEBS Lett 584:1956-65. 2010
    ..In this review we summarize the RyR mouse models and how they have enhanced our understanding of the RyR channels and their roles in cellular physiology and disease...
  2. pmc Dysfunctional ryanodine receptors in the heart: new insights into complex cardiovascular diseases
    Steven O Marx
    Division of Cardiology, College of Physicians and Surgeons of Columbia University, New York, NY 10032, USA
    J Mol Cell Cardiol 58:225-31. 2013
    ..Correcting these defects using either genetic manipulation (knock-in) in mice, or specific and novel small molecules ameliorates the RyR2 dysfunction, reducing the progression to heart failure and the incidence of arrhythmias...
  3. pmc Ryanodine receptor channelopathies
    Matthew J Betzenhauser
    Department of Physiology, Clyde and Helen Wu Center for Molecular Cardiology, Columbia University College of Physicians and Surgeons, 630 West 168th Street, New York, NY 10032, USA
    Pflugers Arch 460:467-80. 2010
    ..Thus, RyR channelopathies are both of interest because they cause significant human diseases and provide model systems that can be studied to elucidate important structure-function relationships of these ion channels...