Cardiomyocyte-specific deletion of adhesion G protein-coupled receptor F5 (ADGRF5) increases susceptibility to stress and arrhythmia

Jeanette Einspahr; Viren Patwa; Jacob Melchior; Jhansi Raja; Jing Zhang; Ryan Boudreau; Farah Sheikh

doi:10.1152/physiol.2024.39.S1.1200

Abstract only G-Protein Coupled Receptors (GPCR) represent the largest family of cell surface receptors in the human body and serve as targets for more than 30% of all approved drugs. Adhesion GPCRs (AGPCR) are an understudied GPCR family containing numerous extracellular domains that interact with extracellular proteins, including many known mediators of cardiac remodeling responses to pathologic stress. However, their relative expression in the heart and impact on cardiac function normally or during disease has been largely unstudied. To address this, we evaluated RNASeq-attained AGPCR expression from left ventricular (LV) samples of adult mice, of which Adgrf5 was one of the most highly expressed. Notably, decreased ADGRF5 expression is observed across numerous failing human LV tissue GEO datasets, and C57BL6/J mice subjected to transaortic constriction (TAC) surgery to induce pressure overload-induced HF also displayed reduced LV Adgrf5 expression by 12 weeks post-TAC. Analysis of published single cell (sc)-RNASeq datasets confirmed Adgrf5 expression in cardiomyocytes (CM), which also decreased following TAC in mice and in CM from failing human hearts. To investigate the impact of CM-specific ADGRF5 on cardiac function and remodeling normally or during disease progression, we crossed floxed ADGRF5 (ADGRF5 f/f ) and αMHC-Cre mice to generate constitutive, CM-specific ADGRF5 knockout mice (F5cmKO). These mice display normal cardiac structure and function at 12 weeks of age versus αMHC-Cre mice, as assessed via echocardiography, gravimetrics and immunohistochemistry. However, F5cmKO mice develop cardiac dysfunction, maladaptive remodeling, and increased mortality over time, even in the absence of pathologic insult, suggesting a homeostatic function of CM-specific ADGRF5. Following TAC surgery, F5cmKO mice display similar outcomes including enhanced cardiac dysfunction with accelerated remodeling and increased mortality. To attain mechanistic insight into these outcomes, we performed RNA-sequencing on LV tissue of 12-week-old F5cmKO mice, identifying and subsequently validating Scn1b as significantly upregulated in F5cmKO CM. Scn1b encodes for the β subunits of voltage-gated sodium channels (Scnβ1/β1B), which regulate gating and kinetics of Nav1.5 as well as adhesive CM-CM interactions, disruption of which could lead to arrhythmias. Indeed, conscious electrocardiogram (ECG) readings via telemetry indicated F5cmKO mice had a high burden of premature ventricular contractions (PVC), including ventricular tachycardia associated with cardiac enlargement. Using a combination of luciferase and fluorescence resonance energy transfer (FRET) reporters and gene expression analyses in neonatal rat ventricular myocytes (NRVM), we confirmed that ADGRF5 controls Scn1b expression in a Gα q/11 protein-dependent manner. In all, we have discovered that loss of CM-specific ADGRF5 leads to increased Scn1b expression with enhanced susceptibility to stress, arrythmias and sudden death, thus may provide a novel GPCR target to promote the maintenance of cardiac rhythm. AHA Predoctoral Fellowship 834906 to J.M.E. P01 HL147841 to D.T. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Cardiomyocyte-specific deletion of adhesion G protein-coupled receptor F5 (ADGRF5) increases susceptibility to stress and arrhythmia

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