This is an application for a competitive renewal of our grant R01 HL095780-01. We were originally funded to investigate mechanisms underlying the role of the known central desmosomal component, desmoplakin, in the clinical and cellular features associated with the genetic-based heart disease, arrhythmogenic right ventricular cardiomyopathy (ARVC) that causes sudden death in the young, by generating and characterizing novel desmoplakin deficient model systems. For this application, we uncovered synaptosomal associated protein 29 (SNAP29) as a novel desmoplakin associated protein in the adult human heart in a yeast-two hybrid screen that we show has relevance to ARVC, by leveraging our novel genetic mouse and human cardiac models of ARVC. Although the role of SNAP29 is unknown in the heart, we show that SNAP29 co-localizes to cardiac muscle desmosomal cell-cell junctions in the adult mouse and human heart as well as human induced pluripotent stem cell derived cardiac cells (hiPSC). Furthermore, SNAP29 localization and/or levels at desmosomal junctions are lost in hearts from our mouse model of ARVC and ARVC hiPSC-derived cardiac cells that exhibit striking desmosomal defects and arrhythmogenic behavior. Generation of novel SNAP29 deficient mouse models (global and cardiomyocyte-specific) revealed striking cardiac defects including (i) autophagic defects at the cardiac muscle cell junction (accumulation of autophagic/lysosomal markers and structures at the cell junction) that specifically impacted desmosomal protein levels and (ii) cardiac morphology defects. Data from our mouse model of ARVC provides validation to this mechanism as we reveal that their hearts exhibit similar defects in autophagic control at the cardiac muscle cell junction. We hypothesize that SNAP29 regulates desmosomal protein levels and function in cardiac muscle by controlling desmosomal turnover via autophagy-mediated mechanisms and its loss will trigger loss of desmosomal protein levels as well as function and ultimately cause ARVC.
We aim to determine: (i) the functional role of SNAP29 in the heart by characterizing SNAP29 loss of function mouse models, (ii) the relevance of SNAP29-DSP interaction in human ARVC and cardiomyocytes by expressing human ARVC mutations and using hiPSCs as a tool, and (iii) the SNAP29-dependent events in autophagy that control desmosomal levels/turnover, by analyzing defects in cardiac autophagy (induction and flux) and relevant desmosome targets using SNAP29 loss of function models and overexpression of SNAP29 in an in vitro ARVC model.

Public Health Relevance

Arrhythmogenic right ventricular cardiomyopathy (ARVC) is a cardiac disease of the desmosome; however, only 40% of patients carry genetic mutations in known desmosomal genes, suggesting that novel players remain to be uncovered in the field. We propose to use state-of-the-art molecular, genetic, cellular, and physiological approaches to dissect novel functions of a SNARE family protein in cardiac desmosomal biology and ARVC, based on uncovering its novel interaction with the cardiac desmosomal complex. Our studies will provide novel insights on desmosomal biology in cardiac muscle, while revealing new underlying causes of ARVC that may also account for underlying defects in the remaining 60% of ARVC patients.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
2R01HL095780-06
Application #
8884263
Study Section
Myocardial Ischemia and Metabolism Study Section (MIM)
Program Officer
Lathrop, David A
Project Start
2009-04-01
Project End
2019-03-31
Budget Start
2015-04-01
Budget End
2016-03-31
Support Year
6
Fiscal Year
2015
Total Cost
Indirect Cost
Name
University of California San Diego
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Liang, Yan; Bradford, William H; Zhang, Jing et al. (2018) Four and a half LIM domain protein signaling and cardiomyopathy. Biophys Rev :
Krysiak, Judith; Unger, Andreas; Beckendorf, Lisa et al. (2018) Protein phosphatase 5 regulates titin phosphorylation and function at a sarcomere-associated mechanosensor complex in cardiomyocytes. Nat Commun 9:262
Bradford, William H; Omens, Jeffrey H; Sheikh, Farah (2017) Vinculin at the heart of aging. Ann Transl Med 5:62
Zanella, Fabian; Sheikh, Farah (2016) Patient-Specific Induced Pluripotent Stem Cell Models: Generation and Characterization of Cardiac Cells. Methods Mol Biol 1353:147-62
Pellman, Jason; Zhang, Jing; Sheikh, Farah (2016) Myocyte-fibroblast communication in cardiac fibrosis and arrhythmias: Mechanisms and model systems. J Mol Cell Cardiol 94:22-31
Ma, Xuanyi; Qu, Xin; Zhu, Wei et al. (2016) Deterministically patterned biomimetic human iPSC-derived hepatic model via rapid 3D bioprinting. Proc Natl Acad Sci U S A 113:2206-11
Liang, Yan; Sheikh, Farah (2016) Scaffold Proteins Regulating Extracellular Regulated Kinase Function in Cardiac Hypertrophy and Disease. Front Pharmacol 7:37
Mezzano, Valeria; Liang, Yan; Wright, Adam T et al. (2016) Desmosomal junctions are necessary for adult sinus node function. Cardiovasc Res 111:274-86
Lyon, Robert C; Zanella, Fabian; Omens, Jeffrey H et al. (2015) Mechanotransduction in cardiac hypertrophy and failure. Circ Res 116:1462-1476
Hribar, K C; Finlay, D; Ma, X et al. (2015) Nonlinear 3D projection printing of concave hydrogel microstructures for long-term multicellular spheroid and embryoid body culture. Lab Chip 15:2412-8

Showing the most recent 10 out of 30 publications