Autophagy is a protective mechanism of cardiomyocytes. Hyperhomocysteinemia (HHcy) elevates oxidative and nitrosative stress levels, leading to an abnormal increase in nitration protein, possibly leading to abnormal autophagy regulation in cardiomyocytes. However, the regulatory effect of HHcy on autophagy at the post-translational modification level is still unclear. Here, we aimed to explore the regulatory mechanism of HHcy on transcription factor EB (TFEB) and nitration of CCAAT/enhancer-binding protein beta (C/EBPβ), a transcriptional repressor of Tfeb, on autophagy in cardiomyocytes.
Results:
In this study, we established the HHcy rat model by feeding a 2.5% (w/w) methionine diet. The nitration level of C/EBPβ was increased in HHcy, which promoted the entry of C/EBPβ into the nucleus, enhanced the transcriptional suppressive effect of C/EBPβ on Tfeb, and induced insufficient autophagy in cardiomyocytes. Furthermore, we confirmed that the Tyr 274 site of C/EBPβ could undergo nitration induced by HHcy. Once C/EBPβ was nitrated on the Tyr 274 site, the nuclear translocation of C/EBPβ and transcription suppressor function of C/EBPβ on Tfeb were enhanced.
Innovation and Conclusion:
We find that C/EBPβ is a transcriptional repressor of Tfeb, and HHcy induces the nitration at the Tyr 274 site of C/EBPβ, leading to autophagic flux blockage in cardiomyocytes. These data indicated that nitrated C/EBPβ might be a potential therapeutic target against HHcy-induced autophagy insufficiency of cardiomyocytes. Antioxid. Redox Signal. 42, 165–183.
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References
1.
AbokyiS, Ghartey-KwansahG, TseDY. TFEB is a central regulator of the aging process and age-related diseases. Ageing Res Rev, 2023; 89:101985; doi: 10.1016/j.arr.2023.101985
2.
Au-YeungKK, WooCW, SungFL, et al.Hyperhomocysteinemia activates nuclear factor-kappaB in endothelial cells via oxidative stress. Circ Res, 2004; 94(1):28–36; doi: 10.1161/01.RES.0000108264.67601.2C
3.
BarakatDJ, MendoncaJ, BarberiT, et al.C/EBPbeta regulates sensitivity to bortezomib in prostate cancer cells by inducing REDD1 and autophagosome-lysosome fusion. Cancer Lett, 2016; 375(1):152–161; doi: 10.1016/j.canlet.2016.03.005
4.
ChenD, WangZ, ZhaoYG, et al.Inositol polyphosphate multikinase inhibits liquid-liquid phase separation of TFEB to negatively regulate autophagy activity. Dev Cell, 2020; 55(5):588–602.e7; doi: 10.1016/j.devcel.2020.10.010
5.
ChenD, YuW, ZhongC, et al.Elabela ameliorates doxorubicin-induced cardiotoxicity by promoting autophagic flux through TFEB pathway. Pharmacol Res, 2022; 178:106186; doi: 10.1016/j.phrs.2022.106186
6.
CzibikG, MezdariZ, MuratAD, et al.Dysregulated phenylalanine catabolism plays a key role in the trajectory of cardiac aging. Circulation, 2021; 144(7):559–574; doi: 10.1161/CIRCULATIONAHA.121.054204
7.
DengY, LiZ, AnX, et al.Hyperhomocysteinemia promotes cardiac hypertrophy in hypertension. Oxid Med Cell Longev, 2022; 2022:1486157; doi: 10.1155/2022/1486157eCollection 2022
8.
DewanjeeS, VallamkonduJ, KalraRS, et al.Autophagy in the diabetic heart: A potential pharmacotherapeutic target in diabetic cardiomyopathy. Ageing Res Rev, 2021; 68:101338; doi: 10.1016/j.arr.2021.101338
9.
FengN, YuH, WangY, et al.Exercise training attenuates angiotensin II-induced cardiac fibrosis by reducing POU2F1 expression. J Sport Health Sci, 2023; 12(4):464–476; doi: 10.1016/j.jshs.2022.10.004
10.
FengQ, LuoY, ZhangXN, et al.MAPT/Tau accumulation represses autophagy flux by disrupting IST1-regulated ESCRT-III complex formation: A vicious cycle in Alzheimer neurodegeneration. Autophagy, 2020; 16(4):641–658; doi: 10.1080/15548627.2019.1633862
11.
GiamoganteF, BarazzuolL, MaiorcaF, et al.A SPLICS reporter reveals [Formula: See text]-synuclein regulation of lysosome-mitochondria contacts which affects TFEB nuclear translocation. Nat Commun, 2024; 15(1):1516; doi: 10.1038/s41467-024-46007-2
12.
HerrmannM, Taban-ShomalO, HubnerU, et al.A review of homocysteine and heart failure. Eur J Heart Fail, 2006; 8(6):571–576; doi: 10.1016/j.ejheart.2005.11.016
13.
HerrmannW, HerrmannM. The controversial role of HCY and Vitamin B deficiency in cardiovascular diseases. Nutrients, 2022; 14(7); doi: 10.3390/nu14071412
14.
HuangX, LiD, ZhaoQ, et al.Association between BHMT and CBS gene promoter methylation with the efficacy of folic acid therapy in patients with hyperhomocysteinemia. J Hum Genet, 2019; 64(12):1227–1235; doi: 10.1038/s10038-019-0672-7
15.
KaplanP, TatarkovaZ, SivonovaMK, et al.Homocysteine and mitochondria in cardiovascular and cerebrovascular systems. Int J Mol Sci, 2020; 21(20):7698; doi: 10.3390/ijms21207698
LambooyS, HeidaA, JoschkoC, et al.Selective hepatic Cbs knockout aggravates liver damage, endothelial dysfunction and ROS stress in mice fed a western diet. Int J Mol Sci, 2023; 24(8):7019; doi: 10.3390/ijms24087019
18.
LintonPJ, GurneyM, SengstockD, et al.This old heart: Cardiac aging and autophagy. J Mol Cell Cardiol, 2015; 83:44–54; doi: 10.1016/j.yjmcc.2014.12.017
19.
MengZ, ZhangZ, ZhaoJ, et al.Nitrative modification of caveolin-3: A novel mechanism of cardiac insulin resistance and a potential therapeutic target against ischemic heart failure in prediabetic animals. Circulation, 2023; 147(15):1162–1179; doi: 10.1161/CIRCULATIONAHA.122.063073
20.
PalomerX, Roman-AzconaMS, Pizarro-DelgadoJ, et al.SIRT3-mediated inhibition of FOS through histone H3 deacetylation prevents cardiac fibrosis and inflammation. Signal Transduct Target Ther, 2020; 5(1):14; doi: 10.1038/s41392-020-0114-1
21.
SciarrettaS, MaejimaY, ZablockiD, et al.The role of autophagy in the heart. Annu Rev Physiol, 2018; 80:1–26; doi: 10.1146/annurev-physiol-021317-121427
22.
SteedMM, TyagiN, SenU, et al.Functional consequences of the collagen/elastin switch in vascular remodeling in hyperhomocysteinemic wild-type, eNOS-/-, and iNOS-/- mice. Am J Physiol Lung Cell Mol Physiol, 2010; 299(3):L301–L311; doi: 10.1152/ajplung.00065.2010
23.
SteedMM, TyagiSC. Mechanisms of cardiovascular remodeling in hyperhomocysteinemia. Antioxid Redox Signal, 2011; 15(7):1927–1943; doi: 10.1089/ars.2010.3721
24.
SunZ, FangC, XuS, et al.SIRT3 attenuates doxorubicin-induced cardiotoxicit-y by inhibiting NLRP3 inflammasome via autophagy. Biochem Pharmacol, 2023; 207:115354; doi: 10.1016/j.bcp.2022.115354
25.
TanA, PrasadR, LeeC, et al.Past, present, and future perspectives of transcription factor EB (TFEB): Mechanisms of regulation and association with disease. Cell Death Differ, 2022; 29(8):1433–1449; doi: 10.1038/s41418-022-01028-6
26.
TangY, LiuY, ZhouH, et al.Esketamine is neuroprotective against traumati brain injury through its modulation of autophagy and oxidative stress via AMPK/mTOR-dependent TFEB nuclear translocation. Exp Neurol, 2023; 366:114436; doi: 10.1016/j.expneurol.2023.114436
27.
TianH, TianD, ZhangC, et al.Efficacy of folic acid therapy in patients with hyperhomocysteinemia. J Am Coll Nutr, 2017; 36(7):528–532; doi: 10.1080/07315724.2017.1330162
28.
TopalG, BrunetA, MillanvoyeE, et al.Homocysteine induces oxidative stressby uncoupling of NO synthase activity through reduction of tetrahydrobiopterin. Free Radic Biol Med, 2004; 36(12):1532–1541; doi: 10.1016/j.freeradbiomed.2004.03.019
29.
TripathiM, SinghBK, ZhouJ, et al.Vitamin B(12) and folate decrease inflammation and fibrosis in NASH by preventing syntaxin 17 homocysteinylation. J Hepatol, 2022; 77(5):1246–1255; doi: 10.1016/j.jhep.2022.06.033
30.
VacekTP, VacekJC, TyagiSC. Mitochondrial mitophagic mechanisms of myocardial matrix metabolism and remodeling. Arch Physiol Biochem, 2012; 118(1):31–42; doi: 10.3109/13813455.2011.635660
31.
WangH, SunQ, ZhouY, et al.Nitration-mediated deficiency of cystathionine beta-synthase activity accelerates the progression of hyperhomocysteinemia. Free Radic Biol Med, 2017; 113:519–529; doi: 10.1016/j.freeradbiomed.2017.10.389
32.
WangMN, LiuLX, DengYT, et al.Regulatory mechanism of heat shock protein 90 on autophagy-related transcription factor EB in human hepatocellular carcinoma cells. Sheng Li Xue Bao, 2020; 72(2):157–166.
33.
WangQ, FuC, XiaH, et al.Elevated plasma homocysteine level associated with further left ventricular structure and function damages in type 2 diabetic patients: A three-dimensional speckle tracking echocardiography study. Metab Syndr Relat Disord, 2021; 19(8):443–451; doi: 10.1089/met.2020.0142
34.
WenW, ZhengH, LiW, et al.Transcription factor EB: A potential integratednetwork regulator in metabolic-associated cardiac injury. Metabolism, 2023; 147:155662; doi: 10.1016/j.metabol.2023.155662
35.
XueK, ChenS, ChaiJ, et al.Nitration of CREB participates in MI-induced myocardial fibrosis via accelerating transcription of col1a2 and cxcl12. Antioxid Redox Signal, 2023; 38(10–12):709–730; doi: 10.1089/ars.2021.0273
36.
YanW, CaoY, ZhenP, et al.Decreased autophagy of vascular smooth muscle cells was involved in hyperhomocysteinemia-induced vascular ageing. Clin Exp Pharmacol Physiol, 2021; 48(4):524–533; doi: 10.1111/1440-1681.13442
37.
YangA, JiaoY, YangS, et al.Homocysteine activates autophagy by inhibition of CFTR expression via interaction between DNA methylation and H3K27me3 in mouse liver. Cell Death Dis, 2018; 9(2):169; doi: 10.1038/s41419-017-0216-z
38.
YangS, NieT, SheH, et al.Regulation of TFEB nuclear localization by HSP90AA1 promotes autophagy and longevity. Autophagy, 2023; 19(3):822–838; doi: 10.1080/15548627.2022.2105561
39.
YuY, ZhaoN, AnJ, et al.CCAAT/enhancer-binding protein beta mediates the killing of toxoplasma gondii by inducing autophagy in nonhematopoietic cells. DNA Cell Biol, 2017; 36(3):212–218; doi: 10.1089/dna.2016.3434
40.
ZahidM, RogowskiM, PonceC, et al.CCAAT/enhancer-binding protein beta (C/EBP beta) knockdown reduces inflammation, ER stress, and apoptosis, and promotes autophagy in oxLDL-treated RAW264.7 macrophage cells. Mol Cell Biochem, 2020; 463(1–2):211–223; doi: 10.1007/s11010-019-03642-4
41.
ZaricBL, ObradovicM, BajicV, et al.Homocysteine and hyperhomocysteinaemia. Curr Med Chem, 2019; 26(16):2948–2961; doi: 10.2174/0929867325666180313105949
42.
ZhangJW, MaYM, JingL, et al.Synaptic remodeling and reduced expression of the transcription factors, HES1 and HES5, in the cortex neurons of cognitively impaired hyperhomocysteinemic mice. Pathol Res Pract, 2020; 216(6):152953; doi: 10.1016/j.prp.2020.152953
43.
ZhangS, ZhangY, CaoY, et al.Homocysteine induces mitochondrial injury via inhibiting myocardial mitophagy. J. Med. Sci. Cent. South China, 2020b;48(01):9–13; doi: 10.15972/j.cnki.43-1509/r.2020.01.003
44.
ZhangS, ZhangY, ZhangX, et al.Nitrative stress-related autophagic insufficiency participates in hyperhomocysteinemia-induced renal aging. Oxid Med Cell Longev, 2020a;2020:4252047; doi: 10.1155/2020/4252047
45.
ZhangX, JiR, LiaoX, et al.MicroRNA-195 regulates metabolism in failing myocardium via alterations in Sirtuin 3 expression and mitochondrial protein acetylation. Circulation, 2018; 137(19):2052–2067; doi: 10.1161/CIRCULATIONAHA.117.030486
46.
ZhaoJ, LiZ, HouC, et al.Gender differences in risk factors for high plasmahomocysteine levels based on a retrospective checkup cohort using a generalized estimating equation analysis. Lipids Health Dis, 2021; 20(1):31; doi: 10.1186/s12944-021-01459-z
47.
ZhouJ, LuY, LinY, et al.Overexpression of hepatic pescadillo 1 in obesity induces lipid dysregulation by inhibiting autophagy. Transl Res, 2023a;258:1–15; doi: 10.1016/j.trsl.2023.02.003
48.
ZhouY, WangXC, WeiJH, et al.Soluble epoxide hydrolase and TRPC3 channels jointly contribute to homocysteine-induced cardiac hypertrophy: Interrelation and regulation by C/EBPbeta. Biochim Biophys Acta Mol Basis Dis, 2023b;1869(4):166643; doi: 10.1016/j.bbadis.2023.166643
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