# Structure-function analysis for elucidating pathogenicity of cardiac ryanodine receptor genetic variants

> **NIH NIH R01** · COLUMBIA UNIVERSITY HEALTH SCIENCES · 2021 · $770,600

## Abstract

Project summary
 The research proposed in this application is designed to elucidate the structure-function relationships
of a form of exercise-induced sudden death known as catecholaminergic polymorphic ventricular
tachycardia (CPVT), caused by mutations in the Type-2 ryanodine receptor (RyR2)/calcium release
channel. RyR2 channels are required for the release of calcium (Ca2+) from intracellular stores, a process
that triggers cellular functions including excitation-contraction (EC) coupling in the cardiac muscle. RyR2,
along with RyR1 and RyR3, are the largest known ion channels, comprised of the four identical ~565 kDa
channel-forming protomers, as well as regulatory subunits, enzymes, and their respective
targeting/anchoring proteins in a macromolecular complex that exceeds three million daltons. It is known
that RyR2 mutations cause arrhythmias including exercise-induced sudden death, or CPVT, and stress-
induced post-translational modifications of RyR2 contribute to both CPVT and heart failure progression.
The applicants have recently obtained near-atomic-level resolution cryo-electron microscopy (cryo-EM)
reconstructions of Type-1 RyR (RyR1) from highly purified rabbit skeletal muscle in both the closed and the
open states, defining the transmembrane pore in unprecedented detail and placing all cytosolic domains as
tertiary folds, including a Ca2+ domain. Using modeling software, the structure of RyR2 has been modeled
based on homology with RyR1. We propose to study the localization, structural effects, and function of at
least 11 representative pathogenic CPVT mutations, in order to develop a system for understanding how
pathogenic genetic variants in different regions of the channel cause clinical disease. These studies will be
conducted by solving cryo-EM structures of mutant RyR2 channels and by functionally testing these
mutations using a novel, high bandwidth, high-throughput lipid bilayer technology developed by our team.
This technology is capable of identifying channel opening events with nanosecond resolution (compared to
current single channel current resolution of millisecond resolution). We will then develop a database of all
known genetic variants CPVT-associated and systematically engineer these mutations into recombinant
RyR2 in order to study these channels using our novel high-throughput lipid bilayer measurement system.
The data from this project will be useful for understanding the underlying mechanisms of CPVT. It will
provide an approach that can be used to develop therapies for CPVT. It will advance our understanding of
novel technologies for studying other diseases caused by RyR2 dysfunction and for studying other ion
channels.

## Key facts

- **NIH application ID:** 10146468
- **Project number:** 5R01HL145473-03
- **Recipient organization:** COLUMBIA UNIVERSITY HEALTH SCIENCES
- **Principal Investigator:** ANDREW Robert MARKS
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $770,600
- **Award type:** 5
- **Project period:** 2019-07-01 → 2023-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10146468

## Citation

> US National Institutes of Health, RePORTER application 10146468, Structure-function analysis for elucidating pathogenicity of cardiac ryanodine receptor genetic variants (5R01HL145473-03). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10146468. Licensed CC0.

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