This is the revised version of an application for competitive renewal of grant RO1-GM57691-13. Since its inception (in 1999), we focused on the mechanisms that regulate gap junctions formed by Connexin43 (Cx43). For this application, we take the reverse angle, and look at the ability of Cx43 to be the regulator of other molecular complexes. In its classical definition, the function of Connexin43 (Cx43) is to form gap junctions. While this description is certainly correct, it does not exclude the possibility that Cx43 exerts other actions, separate from that of gap junction pore formation. Here, we propose that a) specific amino acids within the Cx43 carboxyl terminal domain modulate the function of sodium channels, b) this regulation occurs within the confines of the "perinexus," that is, the area surrounding a gap junction plaque, and c) Cx43-mediated regulation of cell electrophysiology is necessary for proper cardiac rhythm.
Under Aim 1, we will characterize the cardiac electrophysiological profile of mice after loss of Cx43 expression (Cx43-CKO), or after deletion of the last five amino acids of Cx43 (line Cx43D378stop). Both of these mice present with ventricular fibrillation and sudden death. A fundamental difference is that mutation D378stop does not prevent formation of gap junction plaques. We propose that Cx43 is necessary for proper function of other ion channels.
In aim 2, we will define the structure of the perinexus in relation to the sodium channel complex, and the importance of preservation of region 378-382 of Cx43 to control the distance between molecules. We implement a unique combination of modern imaging/recording techniques (scanning ion conductance microscopy, SICM-guided patch clamp, tomographic electron microscopy, direct stochastic reconstruction microscopy, proximity ligation assays), and novel animal models, to address new hypotheses that directly impact our understanding of the molecular mechanisms of cardiac arrhythmias. Overall, our experiments challenge the prevailing concept that, in a structurally normal heart, Cx43-dependent arrhythmias are only consequent to the loss of gap junction channels between cells.

Public Health Relevance

Arrhythmias are a major cause of morbidity and mortality, at great economic and societal cost. A role of Cx43 in the function of non---junctional ion channels remains largely unexplored. Yet, Cx43 is seen as a potential target for arrhythmia therapy, and Cx43 remodeling is considered an important arrhythmia substrate. We seek to shift the paradigm, from the perception of Cx43 as a single---function molecule, to that of a multi---tasking component of a protein interacting network that includes the sodium channel complex at the cardiac intercalated disc.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM057691-16
Application #
8668067
Study Section
Electrical Signaling, Ion Transport, and Arrhythmias Study Section (ESTA)
Program Officer
Nie, Zhongzhen
Project Start
1999-05-01
Project End
2017-04-30
Budget Start
2014-05-01
Budget End
2015-04-30
Support Year
16
Fiscal Year
2014
Total Cost
$410,190
Indirect Cost
$168,190
Name
New York University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
121911077
City
New York
State
NY
Country
United States
Zip Code
10016
Yin, Yandong; Rothenberg, Eli (2016) Probing the Spatial Organization of Molecular Complexes Using Triple-Pair-Correlation. Sci Rep 6:30819
Corrado, Domenico; Zorzi, Alessandro; Cerrone, Marina et al. (2016) Relationship Between Arrhythmogenic Right Ventricular Cardiomyopathy and Brugada Syndrome: New Insights From Molecular Biology and Clinical Implications. Circ Arrhythm Electrophysiol 9:e003631
Shekhar, Akshay; Lin, Xianming; Liu, Fang-Yu et al. (2016) Transcription factor ETV1 is essential for rapid conduction in the heart. J Clin Invest 126:4444-4459
Leo-Macias, Alejandra; Agullo-Pascual, Esperanza; Delmar, Mario (2016) The cardiac connexome: Non-canonical functions of connexin43 and their role in cardiac arrhythmias. Semin Cell Dev Biol 50:13-21
Leo-Macias, Alejandra; Agullo-Pascual, Esperanza; Sanchez-Alonso, Jose L et al. (2016) Nanoscale visualization of functional adhesion/excitability nodes at the intercalated disc. Nat Commun 7:10342
Dubash, Adi D; Kam, Chen Y; Aguado, Brian A et al. (2016) Plakophilin-2 loss promotes TGF-β1/p38 MAPK-dependent fibrotic gene expression in cardiomyocytes. J Cell Biol 212:425-38
Delmar, Mario; Morley, Gregory E (2015) Genetically Encoded Voltage Indicators: Mapping Cardiac Electrical Activity Under a New Light. Circ Res 117:390-1
Lin, Xianming; O'Malley, Heather; Chen, Chunling et al. (2015) Scn1b deletion leads to increased tetrodotoxin-sensitive sodium current, altered intracellular calcium homeostasis and arrhythmias in murine hearts. J Physiol 593:1389-407
Young, Lauren M; Marzio, Antonio; Perez-Duran, Pablo et al. (2015) TIMELESS Forms a Complex with PARP1 Distinct from Its Complex with TIPIN and Plays a Role in the DNA Damage Response. Cell Rep 13:451-9
Leo-Macías, Alejandra; Liang, Feng-Xia; Delmar, Mario (2015) Ultrastructure of the intercellular space in adult murine ventricle revealed by quantitative tomographic electron microscopy. Cardiovasc Res 107:442-52

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