Accumulation of newly synthesized docosahexaenoic acid plays an essential role in heart regeneration

Abstract

Adult zebrafish and neonatal mice can fully regenerate their hearts after partial amputation through the proliferation of preexisting cardiomyocytes (CMs). However, the adult mammalian heart has limited regenerative capability following cardiac damage. The reason for this phenomenon remains elusive. Here, we find that docosahexaenoic acid (DHA) is accumulated only in the injured hearts of zebrafish and neonatal mice, but not of adult mice, which coincides with the upregulation of DHA synthesis genes in CMs, fibroblasts, and macrophages near the injury areas. Inhibition of Fads2, a DHA synthesis enzyme, impairs heart regeneration in both zebrafish and neonatal mice. Injection of DHA remodels the transcriptome from injury response to regeneration response and improves cardiac function in adult mice after myocardial infarction. Interestingly, DHA facilitates CM proliferation but inhibits fibrosis and inflammation. Mechanistically, only DHA, but not oleic acid (OA), can trigger the peroxisome proliferator-activated receptor d (PPARD) to bind to the promoter regions of heart regeneration-related genes, such as Mef2d, Phlda3, and Txndc5, to regulate their expression. Molecular docking, molecular dynamics simulations, and mutagenesis experiments suggest that DHA binds to PPARD in a distinct manner compared to OA, which may help explain their differing abilities to influence the expression of heart regeneration genes. Our findings demonstrate that the DHA signal plays an essential and evolutionarily conserved role in heart regeneration and provide a therapeutic potential for myocardial infarction.

Graphical abstract

Keywords

heart regeneration / zebrafish / mouse / DHA / PPARD

References

[1] Ajith TA , Jayakumar TG. Omega-3 fatty acids in coronary heart disease: recent updates and future perspectives. Clin Exp Pharmacol Physiol 2019; 46: 11– 18.

[2] Cardoso AC , Lam NT , Savla JJ et al. Mitochondrial substrate utilization regulates cardiomyocyte cell cycle progression. Nat Metab 2020; 2: 167– 178.

[3] Chablais F , Jazwinska A. The regenerative capacity of the zebrafish heart is dependent on TGFbeta signaling. Development 2012; 139: 1921– 1930.

[4] Chen J , Ng SM , Chang C et al. p53 isoform delta113p53 is a p53 target gene that antagonizes p53 apoptotic activity via BclxL activation in zebrafish. Genes Dev 2009; 23: 278– 290.

[5] Chen Y , Luttmann FF , Schoger E et al. Reversible reprogramming of cardiomyocytes to a fetal state drives heart regeneration in mice. Science 2021; 373: 1537– 1540.

[6] Cheng X , Ju J , Huang W et al. cpt1b regulates cardiomyocyte proliferation through modulation of glutamine synthetase in zebrafish. J Cardiovasc Dev Dis 2024; 11: 344.

[7] D’Souza K , Nzirorera C , Kienesberger PC. Lipid metabolism and signaling in cardiac lipotoxicity. Biochim Biophys Acta 2016; 1861: 1513– 1524.

[8] Dai J , Liang K , Zhao S et al. Chemoproteomics reveals baicalin activates hepatic CPT1 to ameliorate diet-induced obesity and hepatic steatosis. Proc Natl Acad Sci USA 2018; 115: E5896– E5905.

[9] Dietary supplementation with n-3 polyunsaturated fatty acids and vitamin E after myocardial infarction: results of the GISSI-Prevenzione trial. Gruppo Italiano per lo Studio della Sopravvivenza nell’Infarto miocardico. Lancet 1999;354:447–455.

[10] Doenst T , Nguyen TD , Abel ED. Cardiac metabolism in heart failure: implications beyond ATP production. Circ Res 2013; 113: 709– 724.

[11] Engel FB , Schebesta M , Duong MT et al. p38 MAP kinase inhibition enables proliferation of adult mammalian cardiomyocytes. Genes Dev 2005; 19: 1175– 1187.

[12] FitzSimons M , Beauchemin M , Smith AM et al. Cardiac injury modulates critical components of prostaglandin E₂ signaling during zebrafish heart regeneration. Sci Rep 2020; 10: 3095.

[13] Flinn MA , Jeffery BE , O’Meara CC et al. Yap is required for scar formation but not myocyte proliferation during heart regeneration in zebrafish. Cardiovasc Res 2019; 115: 570– 577.

[14] Fukuda R , Marin-Juez R , El-Sammak H et al. Stimulation of glycolysis promotes cardiomyocyte proliferation after injury in adult zebrafish. EMBO Rep 2020; 21: e49752.

[15] Gemberling M , Karra R , Dickson AL et al. Nrg1 is an injury-induced cardiomyocyte mitogen for the endogenous heart regeneration program in zebrafish. ELIFE 2015; 4: e05871.

[16] Gonzalez-Rosa JM , Sharpe M , Field D et al. Myocardial polyploidization creates a barrier to heart regeneration in zebrafish. Dev Cell 2018; 44: 433– 446.e7.

[17] Grygiel-Górniak B. Peroxisome proliferator-activated receptors and their ligands: nutritional and clinical implications – a review. Nutr J 2014; 13: 17.

[18] Han P , Zhou XH , Chang N et al. Hydrogen peroxide primes heart regeneration with a derepression mechanism. Cell Res 2014; 24: 1091– 1107.

[19] Han L , Shen WJ , Bittner S et al. PPARs: regulators of metabolism and as therapeutic targets in cardiovascular disease. Part Ⅱ: PPAR-β/δ and PPAR-γ. Future Cardiol 2017; 13: 279– 296.

[20] Harmon SD , Kaduce TL , Manuel TD et al. Effect of the delta6-desaturase inhibitor SC-26196 on PUFA metabolism in human cells. Lipids 2003; 38: 469– 476.

[21] Hirose K , Payumo AY , Cutie S et al. Evidence for hormonal control of heart regenerative capacity during endothermy acquisition. Science 2019; 364: 184– 188.

[22] Honkoop H , de Bakker DE , Aharonov A et al. Single-cell analysis uncovers that metabolic reprogramming by ErbB2 signaling is essential for cardiomyocyte proliferation in the regenerating heart. eLife 2019; 8: e50163.

[23] Humayun A , Fornace AJ. GADD45 in stress signaling, cell cycle control, and apoptosis. Adv Exp Med Biol 2022; 1360: 1– 22.

[24] Ji X , Chen Z , Wang Q et al. Sphingolipid metabolism controls mammalian heart regeneration. Cell Metab 2024; 36: 839– 856.e8.

[25] Jopling C , Sleep E , Raya M et al. Zebrafish heart regeneration occurs by cardiomyocyte dedifferentiation and proliferation. Nature 2010; 464: 606– 609.

[26] Jopling C , Sune G , Faucherre A et al. Hypoxia induces myocardial regeneration in zebrafish. Circulation 2012a; 126: 3017– 3027.

[27] Jopling C , Sune G , Morera C et al. p38α MAPK regulates myocardial regeneration in zebrafish. Cell Cycle 2012b; 11: 1195– 1201.

[28] Kikuchi K , Holdway JE , Werdich AA et al. Primary contribution to zebrafish heart regeneration by gata4₊ cardiomyocytes. Nature 2010; 464: 601– 605.

[29] Korbecki J , Bobinski R , Dutka M. Self-regulation of the inflammatory response by peroxisome proliferator-activated receptors. Inflamm Res 2019; 68: 443– 458.

[30] Koth J , Wang X , Killen AC et al. Runx1 promotes scar deposition and inhibits myocardial proliferation and survival during zebrafish heart regeneration. Development 2020; 147: dev186569.

[31] Li Y , Li Z , Zhang C et al. Cardiac fibroblast-specific activating transcription factor 3 protects against heart failure by suppressing MAP2K3-p38 signaling. Circulation 2017; 135: 2041– 2057.

[32] Li X , Wu F , Günther S et al. Inhibition of fatty acid oxidation enables heart regeneration in adult mice. Nature 2023; 622: 619– 626.

[33] Lin YH , Zhang S , Zhu M et al. Mice with increased numbers of polyploid hepatocytes maintain regenerative capacity but develop fewer hepatocellular carcinomas following chronic liver injury. Gastroenterology 2020; 158: 1698– 1712.e14.

[34] Magadum A , Ding Y , He L et al. Live cell screening platform identifies PPARdelta as a regulator of cardiomyocyte proliferation and cardiac repair. Cell Res 2017; 27: 1002– 1019.

[35] Maillet M , Purcell NH , Sargent MA et al. DUSP6 (MKP3) null mice show enhanced ERK1/2 phosphorylation at baseline and increased myocyte proliferation in the heart affecting disease susceptibility. J Biol Chem 2008; 283: 31246– 31255.

[36] Meng X , Zhang L , Han B et al. PHLDA3 inhibition protects against myocardial ischemia/reperfusion injury by alleviating oxidative stress and inflammatory response via the Akt/Nrf2 axis. Environ Toxicol 2021; 36: 2266– 2277.

[37] Mukherjee D , Wagh G , Mokalled MH et al. Ccn2a is an injury-induced matricellular factor that promotes cardiac regeneration in zebrafish. Development 2021; 148: dev193219.

[38] Oppedisano F , Macrì R , Gliozzi M et al. The anti-inflammatory and antioxidant properties of n-3 PUFAs: their role in cardiovascular protection. Biomedicines 2020; 8: 306.

[39] Patterson M , Barske L , Van Handel B et al. Frequency of mononuclear diploid cardiomyocytes underlies natural variation in heart regeneration. Nat Genet 2017; 49: 1346– 1353.

[40] Picelli S , Faridani OR , Bjorklund AK et al. Full-length RNA-seq from single cells using Smart-seq2. Nat Protoc 2014; 9: 171– 181.

[41] Piquereau J , Ventura-Clapier R. Maturation of cardiac energy metabolism during perinatal development. Front Physiol 2018; 9: 959.

[42] Porrello ER , Mahmoud AI , Simpson E et al. Transient regenerative potential of the neonatal mouse heart. Science 2011; 331: 1078– 1080.

[43] Poss KD , Wilson LG , Keating MT. Heart regeneration in zebrafish. Science 2002; 298: 2188– 2190.

[44] Schmidt-Schweda S , Holubarsch C. First clinical trial with etomoxir in patients with chronic congestive heart failure. Clin Sci (Lond) 2000; 99: 27– 35.

[45] Shi Y , Tian M , Zhao X et al. α-Ketoglutarate promotes cardiomyocyte proliferation and heart regeneration after myocardial infarction. Nat Cardiovasc Res 2024; 3: 1083– 1097.

[46] Shih YC , Chen CL , Zhang Y et al. Endoplasmic reticulum protein TXNDC5 augments myocardial fibrosis by facilitating extracellular matrix protein folding and redox-sensitive cardiac fibroblast activation. Circ Res 2018; 122: 1052– 1068.

[47] Shoffner A , Cigliola V , Lee N et al. Tp53 suppression promotes cardiomyocyte proliferation during zebrafish heart regeneration. CELL REP 2020; 32: 108089.

[48] Tahara N , Akiyama R , Wang J et al. The FGF-AKT pathway is necessary for cardiomyocyte survival for heart regeneration in zebrafish. Dev Biol 2021; 472: 30– 37.

[49] Tan J , Yang M , Wang H et al. Moderate heart rate reduction promotes cardiac regeneration through stimulation of the metabolic pattern switch. CELL REP 2022; 38: 110468.

[50] Verlengia R , Gorjao R , Kanunfre CC et al. Effects of EPA and DHA on proliferation, cytokine production, and gene expression in Raji cells. Lipids 2004; 39: 857– 864.

[51] von Gise A , Lin Z , Schlegelmilch K et al. YAP1, the nuclear target of Hippo signaling, stimulates heart growth through cardiomyocyte proliferation but not hypertrophy. Proc Natl Acad Sci USA 2012; 109: 2394– 2399.

[52] Wang W , Hu CK , Zeng A et al. Changes in regeneration-responsive enhancers shape regenerative capacities in vertebrates. Science 2020; 369: eaaz3090.

[53] Wang L , Liu J , Wang Z et al. Dexmedetomidine abates myocardial ischemia reperfusion injury through inhibition of pyroptosis via regulation of miR-665/MEF2D/Nrf2 axis. Biomed Pharmacother 2023; 165: 115255.

[54] Wilkinson PD , Delgado ER , Alencastro F et al. The polyploid state restricts hepatocyte proliferation and liver regeneration in mice. Hepatology 2019; 69: 1242– 1258.

[55] Ye S , Zhao T , Zhang W et al. p53 isoform Delta113p53 promotes zebrafish heart regeneration by maintaining redox homeostasis. Cell Death Dis 2020; 11: 568.

[56] Yu P , Song S , Zhang X et al. Downregulation of apoptotic repressor AVEN exacerbates cardiac injury after myocardial infarction. Proc Natl Acad Sci USA 2023; 120: e1992485176.

[57] Zhao Y , Yang G , Wu N et al. Integrated transcriptome and phosphoproteome analyses reveal that fads2 is critical for maintaining body LC-PUFA homeostasis. J Proteomics 2020; 229: 103967.

[58] Zhao Y , Lv H , Yu C et al. Systemic inhibition of mitochondrial fatty acid beta-oxidation impedes zebrafish ventricle regeneration. Biochim Biophys Acta Mol Basis Dis 2024; 1870: 167442.

[59] Zhou X , Zhang C , Wu X et al. Dusp6 deficiency attenuates neutrophil-mediated cardiac damage in the acute inflammatory phase of myocardial infarction. Nat Commun 2022; 13: 6672.

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© The Author(s) 2025. Published by Oxford University Press on behalf of Higher Education Press.