Identification of DNA Damage Repair Enzyme Ascc2 as Causal for Heart Failure With Preserved Ejection Fraction
2022; Lippincott Williams & Wilkins; Volume: 145; Issue: 14 Linguagem: Inglês
10.1161/circulationaha.121.055857
ISSN1524-4539
AutoresYang Cao, Calvin Pan, Yu-Chen Wang, Zhiqiang Zhou, Vida Jedian, Yonghong Meng, Gillian Campbell, Kristina Guardino, Christopher Li, Jessica Wang, Aldons J. Lusis,
Tópico(s)Advancements in Battery Materials
ResumoHomeCirculationVol. 145, No. 14Identification of DNA Damage Repair Enzyme Ascc2 as Causal for Heart Failure With Preserved Ejection Fraction Free AccessLetterPDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissions ShareShare onFacebookTwitterLinked InMendeleyReddit Jump toFree AccessLetterPDF/EPUBIdentification of DNA Damage Repair Enzyme Ascc2 as Causal for Heart Failure With Preserved Ejection Fraction Yang Cao, PhD, Calvin Pan, MS, Yu-Chen Wang, PhD, Zhiqiang Zhou, MD, Vida Jedian, BS, Yonghong Meng, MD, Gillian Campbell, BS, Kristina Guardino, BS, Christopher Li, BS, Jessica Wang, MD, PhD and Aldons J. Lusis, PhD Yang CaoYang Cao Department of Medicine, Division of Cardiology (Y.C., C.P., Y.-C.W., Z.Z., V.J., Y.M., J.W., A.J.L.), University of California, Los Angeles , Calvin PanCalvin Pan Department of Medicine, Division of Cardiology (Y.C., C.P., Y.-C.W., Z.Z., V.J., Y.M., J.W., A.J.L.), University of California, Los Angeles , Yu-Chen WangYu-Chen Wang Department of Medicine, Division of Cardiology (Y.C., C.P., Y.-C.W., Z.Z., V.J., Y.M., J.W., A.J.L.), University of California, Los Angeles , Zhiqiang ZhouZhiqiang Zhou Department of Medicine, Division of Cardiology (Y.C., C.P., Y.-C.W., Z.Z., V.J., Y.M., J.W., A.J.L.), University of California, Los Angeles , Vida JedianVida Jedian Department of Medicine, Division of Cardiology (Y.C., C.P., Y.-C.W., Z.Z., V.J., Y.M., J.W., A.J.L.), University of California, Los Angeles , Yonghong MengYonghong Meng Department of Medicine, Division of Cardiology (Y.C., C.P., Y.-C.W., Z.Z., V.J., Y.M., J.W., A.J.L.), University of California, Los Angeles , Gillian CampbellGillian Campbell Department of Medicine, Division of Cardiology (Y.C., C.P., Y.-C.W., Z.Z., V.J., Y.M., J.W., A.J.L.), University of California, Los Angeles , Kristina GuardinoKristina Guardino Department of Medicine, Division of Cardiology (Y.C., C.P., Y.-C.W., Z.Z., V.J., Y.M., J.W., A.J.L.), University of California, Los Angeles , Christopher LiChristopher Li Department of Medicine, Division of Cardiology (Y.C., C.P., Y.-C.W., Z.Z., V.J., Y.M., J.W., A.J.L.), University of California, Los Angeles , Jessica WangJessica Wang https://orcid.org/0000-0001-7348-0509 Department of Medicine, Division of Cardiology (Y.C., C.P., Y.-C.W., Z.Z., V.J., Y.M., J.W., A.J.L.), University of California, Los Angeles and Aldons J. LusisAldons J. Lusis https://orcid.org/0000-0001-9013-0228 Department of Medicine, Division of Cardiology (Y.C., C.P., Y.-C.W., Z.Z., V.J., Y.M., J.W., A.J.L.), University of California, Los Angeles Human Genetics (A.J.L.), University of California, Los Angeles Microbiology, Immunology, and Molecular Genetics (G.C., K.G., C.L., A.J.L.), University of California, Los Angeles. Originally published4 Apr 2022https://doi.org/10.1161/CIRCULATIONAHA.121.055857Circulation. 2022;145:1102–1104Heart failure with preserved ejection fraction (HFpEF) is an increasingly prevalent syndrome characterized by diastolic dysfunction and preserved ejection fraction, which is distinct from heart failure with reduced ejection fraction in terms of pathogenesis and effective therapeutic management.1 HFpEF is highly heterogeneous and involves multiple genetic and environmental factors, complicating the dissection of genetic mechanisms of the disease. We used systems genetics approaches involving both mouse and human datasets to identify genetic determinants that play causal roles in diastolic function, a prominent feature of HFpEF.MethodsWe initially examined isoproterenol-induced cardiomyopathy in 105 inbred strains of mice constituting the hybrid mouse diversity panel, a systems genetics resource that enables genome-wide association studies (GWAS) in mice.2 Cardiac function was determined weekly using echocardiography and global transcriptomic profiling of left ventricles was performed after 21 days of isoproterenol infusion (Figure [Ai]). We used E/A ratio (the ratio of peak velocity blood flow in early diastole to peak velocity flow in late diastole) as a surrogate for cardiac diastolic function. First, to identify candidate causal genes affecting diastolic function, we used genetic association to map both local loci (within 10 Mb of the gene being regulated) and distal loci controlling gene expression. Those genes whose local expression is correlated with diastolic function are likely to be causal because local variation is largely regulated in cis by neighboring regulatory elements rather than in trans by factors downstream of diastolic function.3 A total of 274 such genes (P<0.01) were identified and are available on request. Second, to rule out false positives and to identify the most promising genes, we examined whether any of these genes mapped to GWAS loci associated with E/A ratio in the isoproterenol hybrid mouse diversity panel (Figure [Aii]) as well as loci identified in human GWAS for traits associated with HFpEF. The Ascc2 gene was 1 of 6 candidates whose cis component was associated with E/A ratio and was also located in both mouse (chromosome 11; peak E/A ratio single nucleotide polymorphism rs29436095; P=1.65E-06; Figure [Aiii]) and human GWAS loci (Figure [B]). Human ASCC2 (activating signal cointegrator 1 complex subunit 2) was originally identified as a component of the ASC-1 (activating signal cointegrator 1) complex. The ubiquitin-binding activity of ASCC2 functions in DNA alkylation damage repair and ribosome quality control pathways.4 In the hybrid mouse diversity panel, Ascc2 is regulated locally (peak single nucleotide polymorphism rs3698210; Figure [Ci]), with the G allele being associated with higher Ascc2 expression and lower E/A ratios (Figure [Cii]). Strains with lower Ascc2 expression exhibited higher E/A ratio (Figure [Ciii]). In addition to E/A ratio, local genetic variation in Ascc2 expression was significantly correlated with several key traits, including lung function (Figure [Civ]). Genes that were significantly associated with Ascc2 in the hybrid mouse diversity panel were consistently enriched in pathways related to DNA damage and repair (Figure [Cv]).Download figureDownload PowerPointFigure. Identification of Ascc2 as a causal gene for heart failure with preserved ejection fraction. A, Identification of Ascc2 as a cis-regulator of diastolic function. Ai, Experimental design for the development of isoproterenol (ISO)–induced cardiomyopathy across hybrid mouse diversity panel (HMDP) strains. Nine-week-old female mice from 105 inbred strains were treated with ISO (30 mg/kg/d) for 21 days (n=3 to 12 for each strain). Heart function was examined weekly with echocardiography (Echo). After 21 days of ISO infusion, mice were killed and left ventricle transcriptome was determined. Aii, Schematic overview showing identification of causal genes for diastolic function. Local (cis) expression quantitative trait loci were determined for a total of 25 697 genes using a 5% false discovery rate and ≈200 000 single nucleotide polymorphisms (SNPs) as described.2Aiii, Heatmap showing the associations between top candidate genes and diastolic traits. B, Human genome-wide association studies (GWAS) loci encompassing the ASCC2 gene were significantly associated with clinical traits related to heart failure with preserved ejection fraction (HFpEF; https://www.ebi.ac.uk/gwas/genes/ASCC2). The human ASCC2 gene exhibits significant local genetic regulation (https://gtexportal.org/home/gene/ASCC2). C, Ascc2 local expression quantitative trait loci maps to a GWAS locus for E/A ratio (the ratio of peak velocity blood flow in early diastole to peak velocity flow in late diastole) in the ISO-HMDP strain and is inversely correlated with diastolic dysfunction. Ci, Manhattan plot showing the significance (–log10 of P) of all SNPs and expression quantitative trait loci of Ascc2 in ISO-HMDP mice. The peak local expression quantitative trait loci SNP for Assc2 expression maps within 10 Kb of the gene and is within the chromosome 11 GWAS locus for E/A ratio.2 The threshold of significance is P<4e-3. Cii, Correlation of E/A ratio and Ascc2 heart expression with genotype at peak SNP associated with Ascc2 on chromosome 11 (rs3698210). Ciii, Ascc2 expression and E/A ratio in indicated strains. Civ, Heatmap showing the correlation between local genetic variation in Ascc2 expression and measures of diastolic function in the ISO-HMDP strain. Cv, Enriched pathways of Ascc2-associated genes (P<0.001) in ISO-HMDP mice. The enrichment analysis was performed by DAVID (Database for Annotation, Visualization and Integrated Discovery). Di, Nppa and Nppb expression in neonatal rat ventricular myocytes treated with phenylephrine (PE; 100 μmol/L) and small interfering RNAs targeting Ascc2 for 24 hours (as compared with *control [CON] and #PE+CON). Dii, Nuclear and cytoplasmic fraction of ASCC2 (activating signal cointegrator 1 complex subunit 2) in neonatal rat ventricular myocytes after PE treatment for 48 hours. Diii, Heart phosphorylated ATM (ataxia telangiectasia mutated) level after high-fat diet + L-NAME (L-NG-Nitro arginine methyl ester) feeding for 7 weeks. The 2-hit HFpEF model was originally developed by Schiattarella et al5 and our results were consistent with the phenotypes they reported. Ei, Quantitative real-time polymerase chain reaction showing Ascc2 mRNA level in heart and other tissues of CON and Ascc2 heterozygous mice (upper panel) and nuclear and cytoplasmic fraction of ASCC2 in the heart after high-fat diet + L-NAME feeding for 7 weeks (lower panel). Eii, Representative echocardiography images (E/A and E/e′ waves), diastolic function, running distance, and glucose tolerance test of CON and Ascc2 heterozygous mice. Procedures for measuring HFpEF phenotypes were essentially identical to those reported by Schiattarella et al.5F, Gene expression and p-ATM levels in the heart of CON and Ascc2 heterozygous mice after high-fat diet + L-NAME feeding. Each point represents a mouse. All data are presented as mean±SEM. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 by 1-way analysis of variance, 2-way analysis of variance, or Student t test. BMI indicates body mass index; FEV1, forced expiratory volume in 1 second; FVC, forced vital capacity; HDL, high-density lipoprotein; HW/TL, heart weight/tibia length; IVS, interventricular septum; LDH, lactate dehydrogenase; LV, left ventricular; and PW, posterior wall.We further validated Ascc2 using in vitro and in vivo models. In neonatal rat ventricular myocytes, Ascc2 knockdown by small interfering RNA caused an increase of Nppa and Nppb expression (Figure [Di]), genes whose expression is increased in patients with HFpEF. The levels of ASCC2 protein localized to the nucleus were increased in neonatal rat ventricular myocytes after treatment with phenylephrine (Figure [Dii]).5 To determine whether HFpEF is associated with an induction of the DNA damage response, we examined ATM (ataxia telangiectasia mutated) kinase, one of the best characterized DNA damage response markers, in a 2-hit HFpEF model induced by feeding a high-fat diet and the nitric oxide inhibitor L-NAME (L-NG-Nitro arginine methyl ester) for 7 weeks.5 Phosphorylated ATM was increased in the heart in the HFpEF mouse model (Figure [Diii]).AnimalsAll animal experiments were approved by the University of California Los Angeles Animal Care and Use Committee in accordance with Public Health Service guidelines.Data AvailabilityThe data that support the findings of this study, including reagents used, statistical methods, gene expression and mapping data, are available from the corresponding author on request.DiscussionWe validated the role of Ascc2 in vivo using cardiac-specific knockout mice generated by breeding Ascc2 flox (C57BL/6 N-Atm1Brd Ascc2tm1a(EUCOMM)Wtsi/Bay Mmucd) with αMHC-Mer-Cre-Mer mice. Control and Ascc2 heterozygous knockout mice were studied in the 2-hit HFpEF model.5 Compared with control mice, Ascc2 heterozygous mice exhibited reduced Ascc2 levels in the heart but not in other tissues. In common with the neonatal rat ventricular myocyte studies, ASCC2 levels in the nucleus increased after feeding mice the HFpEF diet (Figure [Ei]). Ascc2 heterozygous mice exhibited greater diastolic dysfunction and impaired exercise tolerance but retained normal left ventricular ejection fraction and glucose tolerance (Figure [Eii]). Factors associated with heart failure (Nppa and Nppb), DNA damage response (phosopho-ATM), and inflammation (interleukin-6 and tumor necrosis factor) were increased in heart tissue of Ascc2 heterozygous mice (Figure [F]).Our results reveal a role of the DNA damage response in HFpEF. ASCC2 is a component of the ASC-1 complex that plays essential roles in transcriptional regulation, DNA damage repair, and ribosome quality control pathways.4 We hypothesize that ASCC2 translocates into the nucleus in HFpEF and regulates heart function by mediating DNA damage response. The effect appears to occur in cardiomyocytes given our findings in neonatal rat ventricular myocytes and cardiomyocyte-specific knockouts. The fact that the ASCC2 locus is associated with traits relevant to HFpEF in human GWAS suggests that it may play a role in the human disease.Article InformationSources of FundingThis work was supported by National Institutes of Health grants DK120342 and HL147883.Nonstandard Abbreviations and AcronymsASC-1activating signal cointegrator 1ASCC2activating signal cointegrator 1 complex subunit 2ATMataxia telangiectasia mutatedGWASgenome-wide association studiesHFpEFheart failure with preserved ejection fractionL-NAMEL-NG-Nitro arginine methyl esterDisclosures None.FootnotesFor Sources of Funding and Disclosures, see page 1104.Circulation is available at www.ahajournals.org/journal/circCorrespondence to: Aldons J. Lusis, PhD, University of California Los Angeles, MRL 3-730, 675 Charles E Young Drive South, Los Angeles, CA 90095-1679. Email [email protected]ucla.eduReferences1. Dunlay SM, Roger VL, Redfield MM. Epidemiology of heart failure with preserved ejection fraction.Nat Rev Cardiol. 2017; 14:591–602. doi: 10.1038/nrcardio.2017.65CrossrefMedlineGoogle Scholar2. Wang JJ, Rau C, Avetisyan R, Ren S, Romay MC, Stolin G, Gong KW, Wang Y, Lusis AJ. Genetic dissection of cardiac remodeling in an isoproterenol-induced heart failure mouse model.PLoS Genet. 2016; 12:e1006038. doi: 10.1371/journal.pgen.1006038CrossrefMedlineGoogle Scholar3. Seldin M, Yang X, Lusis AJ. Systems genetics applications in metabolism research.Nat Metab. 2019; 1:1038–1050. doi: 10.1038/s42255-019-0132-xCrossrefMedlineGoogle Scholar4. Brickner JR, Soll JM, Lombardi PM, Vågbø CB, Mudge MC, Oyeniran C, Rabe R, Jackson J, Sullender ME, Blazosky E, et al.. A ubiquitin-dependent signalling axis specific for ALKBH-mediated DNA dealkylation repair.Nature. 2017; 551:389–393. doi: 10.1038/nature24484CrossrefMedlineGoogle Scholar5. Schiattarella GG, Altamirano F, Tong D, French KM, Villalobos E, Kim SY, Luo X, Jiang N, May HI, Wang ZV, et al.. Nitrosative stress drives heart failure with preserved ejection fraction.Nature. 2019; 568:351–356. doi: 10.1038/s41586-019-1100-zCrossrefMedlineGoogle Scholar Previous Back to top Next FiguresReferencesRelatedDetails April 5, 2022Vol 145, Issue 14 Advertisement Article InformationMetrics © 2022 American Heart Association, Inc.https://doi.org/10.1161/CIRCULATIONAHA.121.055857PMID: 35377742 Originally publishedApril 4, 2022 Keywordsheart failuregenome-wide association studygeneticsDNA damagePDF download Advertisement SubjectsCardiomyopathyHeart FailureMyocardial Biology
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