The histone methyltransferase SETD2 drives cardiometabolic heart failure with preserved ejection fraction
Authors: Ambrosini S , Mohammed S A , Gorica E , Herwig M , Karsay G , Hornemann T , Ruschitzka F , Hamdani N , Costantino S , Paneni F 
Published: October 2022

Abstract

Background
Heart failure with preserved ejection fraction (HFpEF) is highly prevalent in patients with cardiometabolic disorders and associates with a poor outcome. Pathological gene expression in heart failure is accompanied by changes in active histone marks without major alterations in DNA methylation. Histone 3 trimethylation at lysine 36 (H3k36me3) – an active chromatin mark induced by the methyltransferase SETD2 – was recently found among the top epigenetic signatures in failing human hearts. Yet, the role of SETD2/H3k36me3 in heart failure is poorly understood.

Purpose
To investigate whether SETD2 participates in the transcriptional regulation of cardiometabolic HFpEF.

Methods
Mice with cardiomyocyte-specific deletion of SETD2 (c-SETD2−/−) and control littermates (SETD2fl/fl) were generated and subjected to high fat diet feeding and L-NAME treatment for 15 weeks to induce cardiometabolic HFpEF. Histology, mouse echocardiography (Vevo3100) and Treadmill exhaustion test were performed. ChIP-Seq datasets were employed to determine the biological pathways regulated by H3k36me3, whereas chromatin immunoprecipitation assays (ChIP) were performed to investigate SETD2/H3k36me3 enrichment on gene promoters. SETD2 gain- and loss-of-function experiments were performed in cultured cardiomyocytes (CMs) exposed to palmitic acid. Lipotoxic injury was assessed by mass spectrometry (MS)-based quantification of lipid species, autophagic flux (by Western blot) and apoptosis (by Caspase-3 activity assay). SETD2/H3k36me3 were also investigated in left ventricular myocardial specimens from patients with HFpEF and were correlated to passive stiffness.

Results
ChIP-Seq in mouse CMs showed a strong enrichment of SETD2/H3k36me3 in pathways underpinning triglyceride synthesis. SETD2 and H3k36me3 were upregulated in HFpEF vs. control mouse hearts and were highly enriched on the promoter of sterol regulatory element-binding transcription factor 1 (SREBF1) gene. These changes were associated with SREBF1 upregulation, myocardial triglyceride accumulation and lipotoxic damage. In HFpEF mice, cardiomyocyte-specific deletion of SETD2 prevented hypertrophic remodeling, diastolic dysfunction and lung congestion while improving exercise tolerance. Moreover, SETD2 deletion blunted H3K36me3 enrichment on SREBF1 promoter thus preventing SREBF1-related lipid accumulation, impaired autophagic flux and apoptosis. In cultured CMs exposed to palmitic acid, SETD2 depletion prevented H3k36me3-driven SREBF1 upregulation, whereas SETD2 overexpression recapitulated lipotoxic damage. SREBF1 knockdown prevented lipotoxic injury in SETD2-overexpressing CMs, suggesting its direct role in SETD2 signalling. Finally, SETD2 was upregulated in myocardial samples from obese patients with HFpEF and positively correlated with cardiomyocyte stiffness, a major feature of HFpEF.

Conclusions
SETD2 may represent an attractive molecular target for the prevention of cardiometabolic HFpEF.

Funding Acknowledgement
Type of funding sources: Public Institution(s). Main funding source(s): University of Zürich