Ischemic heart disease and its associated complications of sudden cardiac death and congestive heart failure remain leading causes of morbidity and mortality in the United States. The arrhythmogenic pathophysiology of ischemic disease includes loss of cardiomyocyte electrical coupling through altered localization and modulation of gap junctions. The precise molecular mechanisms underlying altered coupling remain elusive. The objective of this proposal is to understand the cellular movement of gap junction hemichannels (connexons) in normal and ischemic conditions. Our central hypothesis is that connexons require the cytoskeleton to target them to specific locations on the plasma membrane and, once in the plasma membrane, there is a limited role of lateral diffusion and other means of non- cytoskeleton based channel movement. The hypothesis will be tested by using fixed and live cell imaging techniques including high resolution total internal reflection fluorescence (TIRF) imaging, with supplementary biochemistry, to understand the molecular mechanisms of connexon trafficking. Particular aims include understanding the role of microtubule based directed targeting of connexons to intercalated discs in conditions of oxidative stress and simulated ischemia;to understand the role of the actin cytoskeleton in targeted delivery of connexons;and to determine quantitatively the capacity of connexons to diffuse laterally to other membrane regions within the plasma membrane. Preliminary data indicate that oxidative stress limits microtubule capture of cortical membrane, preventing delivery of connexons to the plasma membrane;that actin helps microtubules position and target ion channels to specific regions of the cortical membrane, and that lateral diffusion of connexons is highly restricted.
These aims will further our understanding of the regulation and behavior of cardiac gap junctions. The general field of protein trafficking will benefit from fundamental new knowledge about cytoskeleton and ischemic type regulation of these channels. Furthermore, key molecules and events in the trafficking of gap junctions will be identified to be used as therapeutic targets to lessen the arrhythmias and dysfunction associated with ischemic heart disease.

Public Health Relevance

The clinical sequelae of ischemic heart disease are congestive heart failure and sudden cardiac death which are primary causes of mortality in the United States. The cellular basis of both heart failure and sudden death involve diminished electrical coupling between heart cells. This application proposes to study the molecular mechanisms of electrical coupling between heart cells and identify proteins involved in regulating the coupling under normal conditions and during times of reduced blood flow (ischemia).

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
7R01HL094414-06
Application #
8706447
Study Section
Electrical Signaling, Ion Transport, and Arrhythmias Study Section (ESTA)
Program Officer
Krull, Holly
Project Start
2009-04-15
Project End
2014-03-31
Budget Start
2013-09-01
Budget End
2014-03-31
Support Year
6
Fiscal Year
2013
Total Cost
$344,796
Indirect Cost
$149,523
Name
Cedars-Sinai Medical Center
Department
Type
DUNS #
075307785
City
Los Angeles
State
CA
Country
United States
Zip Code
90048
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Fu, Ying; Shaw, Robin M (2017) CASAAV Technology to Examine Regulators of Heart Failure: Cause or Effect. Circ Res 120:1846-1848
Hong, TingTing; Shaw, Robin M (2017) Cardiac T-Tubule Microanatomy and Function. Physiol Rev 97:227-252
Fu, Ying; Zhang, Shan-Shan; Xiao, Shaohua et al. (2017) Cx43 Isoform GJA1-20k Promotes Microtubule Dependent Mitochondrial Transport. Front Physiol 8:905
Basheer, Wassim A; Xiao, Shaohua; Epifantseva, Irina et al. (2017) GJA1-20k Arranges Actin to Guide Cx43 Delivery to Cardiac Intercalated Discs. Circ Res 121:1069-1080
Basheer, Wassim A; Shaw, Robin M (2016) Connexin 43 and CaV1.2 Ion Channel Trafficking in Healthy and Diseased Myocardium. Circ Arrhythm Electrophysiol 9:e001357
Basheer, Wassim; Shaw, Robin (2016) The ""tail"" of Connexin43: An unexpected journey from alternative translation to trafficking. Biochim Biophys Acta 1863:1848-56
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Fu, Ying; Xiao, Shaohua; Hong, TingTing et al. (2015) Cytoskeleton regulation of ion channels. Circulation 131:689-91

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