Abstract: Oxidative stress and apoptosis are the key factors that limit the hypothermic preservation time of donor hearts to within 4–6 h. The aim of this study was to investigate whether the histone deacetylase 3 (HDAC3) inhibitor RGFP966 could protect against cardiac injury induced by prolonged hypothermic preservation. Rat hearts were hypothermically preserved in Celsior solution with or without RGFP966 for 12 h followed by 60 min of reperfusion. Hemodynamic parameters during reperfusion were evaluated. The expression and phosphorylation levels of mammalian STE20-like kinase-1 (Mst1) and Yes-associated protein (YAP) were determined by western blotting. Cell apoptosis was measured by the terminal deoxynucleotidyl-transferase (TdT)-mediated dUTP nick-end labeling (TUNEL) method. Addition of RGFP966 in Celsior solution significantly inhibited cardiac dysfunction induced by hypothermic preservation. RGFP966 inhibited the hypothermic preservation-induced increase of the phosphorylated (p)-Mst1/Mst1 and p-YAP/YAP ratios, prevented a reduction in total YAP protein expression, and increased the nuclear YAP protein level. Verteporfin (VP), a small molecular inhibitor of YAP–transcriptional enhanced associate domain (TEAD) interaction, partially abolished the protective effect of RGFP966 on cardiac function, and reduced lactate dehydrogenase activity and malondialdehyde content. RGFP966 increased superoxide dismutase, catalase, and glutathione peroxidase gene and protein expression, which was abolished by VP. RGFP966 inhibited hypothermic preservation-induced overexpression of B-cell lymphoma protein 2 (Bcl-2)-associated X (Bax) and cleaved caspase-3, increased Bcl-2 mRNA and protein expression, and reduced cardiomyocyte apoptosis. The antioxidant and anti-apoptotic effects of RGFP966 were cancelled by VP. The results suggest that supplementation of Celsior solution with RGFP966 attenuated prolonged hypothermic preservation-induced cardiac dysfunction. The mechanism may involve inhibition of oxidative stress and apoptosis via inactivation of the YAP pathway.

RGFP966通过抑制YAP通路活化减轻长时程低温保存诱导的心功能损伤

[1]An PP, Li JX, Lu LL, et al., 2019. Histone deacetylase 8 triggers the migration of triple negative breast cancer cells via regulation of YAP signals. Eur J Pharmacol, 845:16-23.

[2]Aune SE, Herr DJ, Mani SK, et al., 2014. Selective inhibition of class I but not class IIb histone deacetylases exerts cardiac protection from ischemia reperfusion. J Mol Cell Cardiol, 72:138-145.

[3]Brundel BJJM, Li J, Zhang DL, 2020. Role of HDACs in cardiac electropathology: therapeutic implications for atrial fibrillation. Biochim Biophys Acta Mol Cell Res, 1867(3):118459.

[4]Chen BY, Jiang LX, Hao K, et al., 2018. Protection of plasma transfusion against lipopolysaccharide/D-galactosamine-induced fulminant hepatic failure through inhibiting apoptosis of hepatic cells in mice. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 19(6):436-444.

[5]Chen GG, Yan JB, Wang XM, et al., 2016. Mechanism of uncoupling protein 2-mediated myocardial injury in hypothermic preserved rat hearts. Mol Med Rep, 14(2):1857-1864.

[6]Chen ML, Liu Q, Chen LJ, et al., 2017. Remifentanil postconditioning ameliorates histone H3 acetylation modification in H9c2 cardiomyoblasts after hypoxia/reoxygenation via attenuating endoplasmic reticulum stress. Apoptosis, 22(5):662-671.

[7]Ferguson BS, McKinsey TA, 2015. Non-sirtuin histone deacetylases in the control of cardiac aging. J Mol Cell Cardiol, 83:14-20.

[8]Fu PF, Zheng X, Fan X, et al., 2019. Role of cytoplasmic lncRNAs in regulating cancer signaling pathways. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 20(1):1-8.

[9]Hamlin RL, del Rio C, 2012. dP/dt_max—a measure of ‘baroinometry’. J Pharmacol Toxicol Methods, 66(2):63-65.

[10]Hitchcock LN, Raybuck JD, Wood MA, et al., 2019. Effects of a histone deacetylase 3 inhibitor on extinction and reinstatement of cocaine self-administration in rats. Psychopharmacology, 236(1):517-529.

[11]Janczura KJ, Volmar CH, Sartor GC, et al., 2018. Inhibition of HDAC3 reverses Alzheimer’s disease-related pathologies in vitro and in the 3xTg-AD mouse model. Proc Natl Acad Sci USA, 115(47):E11148-E11157.

[12]Jia LL, Gu WT, Zhang YP, et al., 2018. Activated Yes-associated protein accelerates cell cycle, inhibits apoptosis, and delays senescence in human periodontal ligament stem cells. Int J Med Sci, 15(11):1241-1250.

[13]Joshi AD, Barabutis N, Birmpas C, et al., 2015. Histone deacetylase inhibitors prevent pulmonary endothelial hyperpermeability and acute lung injury by regulating heat shock protein 90 function. Am J Physiol Lung Cell Mol Physiol, 309(12):L1410-L1419.

[14]Jung DE, Park SB, Kim K, et al., 2017. CG200745, an HDAC inhibitor, induces anti-tumour effects in cholangiocarcinoma cell lines via miRNAs targeting the Hippo pathway. Sci Rep, 7:10921.

[15]Kavitha S, Chandra TS, 2014. Oxidative stress protection and glutathione metabolism in response to hydrogen peroxide and menadione in riboflavinogenic fungus Ashbya gossypii. Appl Biochem Biotechnol, 174(6):2307-2325.

[16]Korkmaz-Icöz S, Li SL, Huttner R, et al., 2019. Hypothermic perfusion of donor heart with a preservation solution supplemented by mesenchymal stem cells. J Heart Lung Transplant, 38(3):315-326.

[17]Lerman DA, Diaz M, Peault B, 2018. Changes in coexpression of pericytes and endogenous cardiac progenitor cells from heart development to disease state. Eur Heart J, 39(S1):P1850.

[18]Lewandowski SL, Janardhan HP, Trivedi CM, 2015. Histone deacetylase 3 coordinates deacetylase-independent epigenetic silencing of transforming growth factor-β1 (TGF-β1) to orchestrate second heart field development. J Biol Chem, 290(45):27067-27089.

[19]Li H, Li XJ, Jing XZ, et al., 2018. Hypoxia promotes maintenance of the chondrogenic phenotype in rat growth plate chondrocytes through the HIF-1α/YAP signaling pathway. Int J Mol Med, 42(6):3181-3192.

[20]Lkhagva B, Lin YK, Kao YH, et al., 2015. Novel histone deacetylase inhibitor modulates cardiac peroxisome proliferator-activated receptors and inflammatory cytokines in heart failure. Pharmacology, 96(3-4):184-191.

[21]Malvaez M, McQuown SC, Rogge GA, et al., 2013. HDAC3-selective inhibitor enhances extinction of cocaine-seeking behavior in a persistent manner. Proc Natl Acad Sci USA, 110(7):2647-2652.

[22]McKinsey TA, 2012. Therapeutic potential for HDAC inhibitors in the heart. Annu Rev Pharmacol Toxicol, 52:303-319.

[23]Mehra MR, Canter CE, Hannan MM, et al., 2016. The 2016 international society for heart lung transplantation listing criteria for heart transplantation: a 10-year update. J Heart Lung Transplant, 35(1):1-23.

[24]Morioka N, Tomori M, Zhang FF, et al., 2016. Stimulation of nuclear receptor REV-ERBs regulates tumor necrosis factor-induced expression of proinflammatory molecules in C6 astroglial cells. Biochem Biophys Res Commun, 469(2):151-157.

[25]Nakamura M, Zhai PY, del Re DP, et al., 2016. Mst1-mediated phosphorylation of Bcl-xL is required for myocardial reperfusion injury. JCI Insight, 1(5):e86217.

[26]Olson DE, Sleiman SF, Bourassa MW, et al., 2015. Hydroxamate-based histone deacetylase inhibitors can protect neurons from oxidative stress via a histone deacetylase-independent catalase-like mechanism. Chem Biol, 22(4):439-445.

[27]Plouffe SW, Lin KC, Moore JL 3rd, et al., 2018. The Hippo pathway effector proteins YAP and TAZ have both distinct and overlapping functions in the cell. J Biol Chem, 293(28):11230-11240.

[28]Poleshko A, Shah PP, Gupta M, et al., 2017. Genome-nuclear lamina interactions regulate cardiac stem cell lineage restriction. Cell, 171(3):573-587.e14.

[29]Ren WB, Xia XJ, Huang J, et al., 2019. Interferon-γ regulates cell malignant growth via the c-Abl/HDAC2 signaling pathway in mammary epithelial cells. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 20(1):39-48.

[30]Rivera-Reyes A, Ye S, Marino GE, et al., 2018. YAP1 enhances NF-κB-dependent and independent effects on clock-mediated unfolded protein responses and autophagy in sarcoma. Cell Death Dis, 9(11):1108.

[31]Roy A, Lordier L, Pioche-Durieu C, et al., 2016. Uncoupling of the Hippo and Rho pathways allows megakaryocytes to escape the tetraploid checkpoint. Haematologica, 101(12):1469-1478.

[32]Ryu Y, Kee HJ, Sun SM, et al., 2019. Class I histone deacetylase inhibitor MS-275 attenuates vasoconstriction and inflammation in angiotensin II-induced hypertension. PLoS ONE, 14(3):e0213186.

[33]Schmitt HM, Schlamp CL, Nickells RW, 2018. Targeting HDAC3 activity with RGFP966 protects against retinal ganglion cell nuclear atrophy and apoptosis after optic nerve injury. J Ocul Pharmacol Ther, 34(3):260-273.

[34]Shao D, Zhai PY, del Re DP, et al., 2014. A functional interaction between Hippo-YAP signalling and FoxO1 mediates the oxidative stress response. Nat Commun, 5(1):3315.

[35]Sharifi-Sanjani M, Shoushtari AH, Quiroz M, et al., 2014. Cardiac CD47 drives left ventricular heart failure through Ca²⁺-CaMKII-regulated induction of HDAC3. J Am Heart Assoc, 3(3):e000670.

[36]Sun Z, Feng D, Fang B, et al., 2013. Deacetylase-independent function of HDAC3 in transcription and metabolism requires nuclear receptor corepressor. Mol Cell, 52(6):769-782.

[37]Wang J, Liu SJ, Heallen T, et al., 2018. The Hippo pathway in the heart: pivotal roles in development, disease, and regeneration. Nat Rev Cardiol, 15(11):672-684.

[38]Xu Z, Tong Q, Zhang ZG, et al., 2017. Inhibition of HDAC3 prevents diabetic cardiomyopathy in OVE26 mice via epigenetic regulation of DUSP5-ERK1/2 pathway. Clin Sci (Lond), 131(15):1841-1857.

[39]Yu LJ, Liu Y, Jin YXZ, et al., 2018. Lentivirus-mediated HDAC3 inhibition attenuates oxidative stress in APPswe/ PS1dE9 mice. J Alzheimers Dis, 61(4):1411-1424.

[40]Zhang DL, Hu X, Li J, et al., 2018. Converse role of class I and class IIa HDACs in the progression of atrial fibrillation. J Mol Cell Cardiol, 125:39-49.

[41]Zhang H, Geng DC, Gao J, et al., 2016. Expression and significance of Hippo/YAP signaling in glioma progression. Tumor Biol, 37(12):15665-15676.

[42]Zhang J, Xu Z, Gu JL, et al., 2018. HDAC3 inhibition in diabetic mice may activate Nrf2 preventing diabetes-induced liver damage and FGF21 synthesis and secretion leading to aortic protection. Am J Physiol Endocrinol Metab, 315(2):E150-E162.

[43]Zhao B, Yuan Q, Hou JB, et al., 2019. Inhibition of HDAC3 ameliorates cerebral ischemia reperfusion injury in diabetic mice in vivo and in vitro. J Diabetes Res, 2019: 8520856.