Spike Protein Impairs Mitochondrial Function in Human Cardiomyocytes: Mechanisms Underlying Cardiac Injury in COVID-19
Abstract
:1. Introduction
2. Materials and Methods
2.1. Human Cardiomyocyte Culture
2.2. Total ATP Assay
2.3. Mitochondrial Bioenergetics
2.4. Mitochondrial Membrane Potential
2.5. Mitochondrial Morphology
2.6. Immunoblotting
2.7. Mitochondrial Ca2+ and Reactive Oxygen Species Levels
2.8. Intracellular Ca2+ Measurements
2.9. Statistical Analysis
3. Results
3.1. S1 Increased Mitochondrial Respiration within 24 h but Impaired Mitochondrial Function within 72 h
3.2. S1 Increased Δψm within 24 h but Disrupted Δψm within 72 h
3.3. S1 Increased Cytosol and Mitochondrial Calcium Levels, and Induced Mitochondrial Fragmentation
3.4. S1 Increased the Expression of Fatty Acid Transporters within 24 h
3.5. S1 Effect Blocked by ACE2 but Not CD147 Neutralization Antibodies
3.6. S1 Increased Mitochondrial ROS Production within 72 h but Not within 24 h
4. Discussion
4.1. Limitations
4.2. Conclusions and Future Perspective
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Mitrani, R.D.; Dabas, N.; Goldberger, J.J. COVID-19 cardiac injury: Implications for long-term surveillance and outcomes in survivors. Heart Rhythm 2020, 17, 1984–1990. [Google Scholar] [CrossRef]
- Psotka, M.A.; Abraham, W.T.; Fiuzat, M.; Filippatos, G.; Lindenfeld, J.; Ahmad, T.; Bhatt, A.S.; Carson, P.E.; Cleland, J.G.; Felker, G.M. Conduct of clinical trials in the era of COVID-19: JACC scientific expert panel. J. Am. Coll. Cardiol. 2020, 76, 2368–2378. [Google Scholar] [CrossRef]
- Raman, B.; Bluemke, D.A.; Lüscher, T.F.; Neubauer, S. Long COVID: Post-acute sequelae of COVID-19 with a cardiovascular focus. Eur. Heart J. 2022, 43, 1157–1172. [Google Scholar] [CrossRef] [PubMed]
- Fox, S.E.; Lameira, F.S.; Rinker, E.B.; Vander Heide, R.S. Cardiac endotheliitis and multisystem inflammatory syndrome after COVID-19. Ann. Intern. Med. 2020, 173, 1025–1027. [Google Scholar] [CrossRef] [PubMed]
- Caforio, A.L.; Baritussio, A.; Basso, C.; Marcolongo, R. Clinically suspected and biopsy-proven myocarditis temporally associated with SARS-CoV-2 infection. Annu. Rev. Med. 2022, 73, 149–166. [Google Scholar] [CrossRef] [PubMed]
- Kawakami, R.; Sakamoto, A.; Kawai, K.; Gianatti, A.; Pellegrini, D.; Nasr, A.; Kutys, B.; Guo, L.; Cornelissen, A.; Mori, M. Pathological evidence for SARS-CoV-2 as a cause of myocarditis: JACC review topic of the week. J. Am. Coll. Cardiol. 2021, 77, 314–325. [Google Scholar] [CrossRef]
- Rhea, E.M.; Logsdon, A.F.; Hansen, K.M.; Williams, L.M.; Reed, M.J.; Baumann, K.K.; Holden, S.J.; Raber, J.; Banks, W.A.; Erickson, M.A. The S1 protein of SARS-CoV-2 crosses the blood-brain barrier in mice. Nat. Neurosci. 2021, 24, 368–378. [Google Scholar] [CrossRef]
- DeOre, B.J.; Tran, K.A.; Andrews, A.M.; Ramirez, S.H.; Galie, P.A. SARS-CoV-2 spike protein disrupts blood-brain barrier integrity via RhoA Activation. J. Neuroimmune Pharmacol. 2021, 16, 722–728. [Google Scholar] [CrossRef]
- Kim, E.S.; Jeon, M.-T.; Kim, K.-S.; Lee, S.; Kim, S.; Kim, D.-G. Spike proteins of SARS-CoV-2 induce pathological changes in molecular delivery and metabolic function in the brain endothelial cells. Viruses 2021, 13, 2021. [Google Scholar] [CrossRef] [PubMed]
- Oh, J.; Cho, W.-H.; Barcelon, E.; Kim, K.H.; Hong, J.; Lee, S.J. SARS-CoV-2 spike protein induces cognitive deficit and anxiety-like behavior in mouse via non-cell autonomous hippocampal neuronal death. Sci. Rep. 2022, 12, 5496. [Google Scholar] [CrossRef]
- Frank, M.G.; Nguyen, K.H.; Ball, J.B.; Hopkins, S.; Kelley, T.; Baratta, M.V.; Fleshner, M.; Maier, S.F. SARS-CoV-2 spike S1 subunit induces neuroinflammatory, microglial and behavioral sickness responses: Evidence of PAMP-like properties. Brain Behav. Immun. 2022, 100, 267–277. [Google Scholar] [CrossRef]
- Avolio, E.; Carrabba, M.; Milligan, R.; Kavanagh Williamson, M.; Beltrami, A.P.; Gupta, K.; Elvers, K.T.; Gamez, M.; Foster, R.R.; Gillespie, K. The SARS-CoV-2 Spike protein disrupts human cardiac pericytes function through CD147 receptor-mediated signalling: A potential non-infective mechanism of COVID-19 microvascular disease. Clin. Sci. 2021, 135, 2667–2689. [Google Scholar] [CrossRef]
- Jennings, R.B.; Ganote, C.E. Mitochondrial structure and function in acute myocardial ischemic injury. Circ. Res. 1976, 38, I80–I91. [Google Scholar] [PubMed]
- Ingwall, J.S.; Weiss, R.G. Is the failing heart energy starved? On using chemical energy to support cardiac function. Circ. Res. 2004, 95, 135–145. [Google Scholar] [CrossRef] [PubMed]
- Weiss, R.G.; Gerstenblith, G.; Bottomley, P.A. ATP flux through creatine kinase in the normal, stressed, and failing human heart. Proc. Natl. Acad. Sci. USA 2005, 102, 808–813. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Tian, R.; Colucci, W.S.; Arany, Z.; Bachschmid, M.M.; Ballinger, S.W.; Boudina, S.; Bruce, J.E.; Busija, D.W.; Dikalov, S.; Dorn, G.W. Unlocking the secrets of mitochondria in the cardiovascular system: Path to a cure in heart failure—A report from the 2018 National Heart, Lung, and Blood Institute Workshop. Circulation 2019, 140, 1205–1216. [Google Scholar] [CrossRef] [PubMed]
- Mercado-Gómez, M.; Prieto-Fernández, E.; Goikoetxea-Usandizaga, N.; Vila-Vecilla, L.; Azkargorta, M.; Bravo, M.; Serrano-Maciá, M.; Egia-Mendikute, L.; Rodríguez-Agudo, R.; Lachiondo-Ortega, S.; et al. The spike of SARS-CoV-2 promotes metabolic rewiring in hepatocytes. Commun. Biol. 2022, 5, 827. [Google Scholar] [CrossRef]
- Huynh, T.V.; Rethi, L.; Chung, C.C.; Yeh, Y.H.; Kao, Y.H.; Chen, Y.J. Class I HDAC modulates angiotensin II-induced fibroblast migration and mitochondrial overactivity. Eur. J. Clin. Investig. 2022, 52, e13712. [Google Scholar] [CrossRef]
- Lkhagva, B.; Kao, Y.-H.; Lee, T.-I.; Lee, T.-W.; Cheng, W.-L.; Chen, Y.-J. Activation of class I histone deacetylases contributes to mitochondrial dysfunction in cardiomyocytes with altered complex activities. Epigenetics 2018, 13, 376–385. [Google Scholar] [CrossRef]
- Yamamoto, H.; Itoh, N.; Kawano, S.; Yatsukawa, Y.; Momose, T.; Makio, T.; Matsunaga, M.; Yokota, M.; Esaki, M.; Shodai, T.; et al. Dual role of the receptor Tom20 in specificity and efficiency of protein import into mitochondria. Proc. Natl. Acad. Sci. USA 2011, 108, 91–96. [Google Scholar] [CrossRef] [Green Version]
- Gabriel, K.; Egan, B.; Lithgow, T. Tom40, the import channel of the mitochondrial outer membrane, plays an active role in sorting imported proteins. EMBO J. 2003, 22, 2380–2386. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ramachandran, K.; Maity, S.; Muthukumar, A.R.; Kandala, S.; Tomar, D.; Abd El-Aziz, T.M.; Allen, C.; Sun, Y.; Venkatesan, M.; Madaris, T.R.; et al. SARS-CoV-2 infection enhances mitochondrial PTP complex activity to perturb cardiac energetics. iScience 2022, 25, 103722. [Google Scholar] [CrossRef]
- Mansanguan, S.; Charunwatthana, P.; Piyaphanee, W.; Dechkhajorn, W.; Poolcharoen, A.; Mansanguan, C. Cardiovascular manifestation of the BNT162b2 mRNA COVID-19 vaccine in adolescents. Trop. Med. Infect. Dis. 2022, 7, 196. [Google Scholar] [CrossRef] [PubMed]
- Yonker, L.M.; Swank, Z.; Bartsch, Y.C.; Burns, M.D.; Kane, A.; Boribong, B.P.; Davis, J.P.; Loiselle, M.; Novak, T.; Senussi, Y.; et al. Circulating spike protein detected in post-COVID-19 mRNA vaccine myocarditis. Circulation 2023. [Google Scholar] [CrossRef]
- Fujioka, Y.; Nishide, S.; Ose, T.; Suzuki, T.; Kato, I.; Fukuhara, H.; Fujioka, M.; Horiuchi, K.; Satoh, A.O.; Nepal, P. A sialylated voltage-dependent Ca2+ channel binds hemagglutinin and mediates influenza A virus entry into mammalian cells. Cell Host Microbe 2018, 23, 809–818.e5. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Hover, S.; Foster, B.; Barr, J.N.; Mankouri, J. Viral dependence on cellular ion channels—An emerging anti-viral target? J. Gen. Virol. 2017, 98, 345–351. [Google Scholar] [CrossRef] [PubMed]
- Wang, K.; Xie, S.; Sun, B. Viral proteins function as ion channels. Biochim. Biophys. Acta 2011, 1808, 510–515. [Google Scholar] [CrossRef] [PubMed]
- Dhont, S.; Derom, E.; Van Braeckel, E.; Depuydt, P.; Lambrecht, B.N. The pathophysiology of ‘happy’ hypoxemia in COVID-19. Respir. Res. 2020, 21, 198. [Google Scholar] [CrossRef]
- Korshunov, S.S.; Skulachev, V.P.; Starkov, A.A. High protonic potential actuates a mechanism of production of reactive oxygen species in mitochondria. FEBS Lett. 1997, 416, 15–18. [Google Scholar] [CrossRef] [Green Version]
- Cheng, J.; Nanayakkara, G.; Shao, Y.; Cueto, R.; Wang, L.; Yang, W.Y.; Tian, Y.; Wang, H.; Yang, X. Mitochondrial proton leak plays a critical role in pathogenesis of cardiovascular diseases. Adv. Exp. Med. Biol. 2017, 982, 359–370. [Google Scholar]
- Liu, S.S. Generating, partitioning, targeting and functioning of superoxide in mitochondria. BioSci. Rep. 1997, 17, 259–272. [Google Scholar] [CrossRef] [PubMed]
- Zhang, H.; Alder, N.N.; Wang, W.; Szeto, H.; Marcinek, D.J.; Rabinovitch, P.S. Reduction of elevated proton leak rejuvenates mitochondria in the aged cardiomyocyte. Elife 2020, 9, e60827. [Google Scholar] [CrossRef] [PubMed]
- Aubert, G.; Vega, R.B.; Kelly, D.P. Perturbations in the gene regulatory pathways controlling mitochondrial energy production in the failing heart. Biochim. Biophys. Acta 2013, 1833, 840–847. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Li, F.; Li, J.; Li, S.; Guo, S.; Li, P. Modulatory effects of Chinese herbal medicines on energy metabolism in ischemic heart diseases. Front. Pharmacol. 2020, 11, 995. [Google Scholar] [CrossRef] [PubMed]
- Franco-Iborra, S.; Cuadros, T.; Parent, A.; Romero-Gimenez, J.; Vila, M.; Perier, C. Defective mitochondrial protein import contributes to complex I-induced mitochondrial dysfunction and neurodegeneration in Parkinson’s disease. Cell Death Dis. 2018, 9, 1122. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Thomas, H.E.; Zhang, Y.; Stefely, J.A.; Veiga, S.R.; Thomas, G.; Kozma, S.C.; Mercer, C.A. Mitochondrial complex I activity is required for maximal autophagy. Cell Rep. 2018, 24, 2404–2417.e8. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lopaschuk, G.D.; Karwi, Q.G.; Tian, R.; Wende, A.R.; Abel, E.D. Cardiac energy metabolism in heart failure. Circ. Res. 2021, 128, 1487–1513. [Google Scholar] [CrossRef]
- Garbincius, J.F.; Elrod, J.W. Is the failing heart starved of mitochondrial calcium? Circ. Res. 2021, 128, 1205–1207. [Google Scholar] [CrossRef]
- Strubbe-Rivera, J.O.; Schrad, J.R.; Pavlov, E.V.; Conway, J.F.; Parent, K.N.; Bazil, J.N. The mitochondrial permeability transition phenomenon elucidated by cryo-EM reveals the genuine impact of calcium overload on mitochondrial structure and function. Sci. Rep. 2021, 11, 1037. [Google Scholar] [CrossRef]
- Lemasters, J.J.; Theruvath, T.P.; Zhong, Z.; Nieminen, A.L. Mitochondrial calcium and the permeability transition in cell death. Biochim. Biophys. Acta 2009, 1787, 1395–1401. [Google Scholar] [CrossRef] [Green Version]
- Lin, L.; Zhang, M.; Yan, R.; Shan, H.; Diao, J.; Wei, J. Inhibition of Drp1 attenuates mitochondrial damage and myocardial injury in Coxsackievirus B3 induced myocarditis. Biochem. Biophys. Res. Commun. 2017, 484, 550–556. [Google Scholar] [CrossRef] [PubMed]
- Liu, H.; Gai, S.; Wang, X.; Zeng, J.; Sun, C.; Zhao, Y.; Zheng, Z. Single-cell analysis of SARS-CoV-2 receptor ACE2 and spike protein priming expression of proteases in the human heart. Cardiovasc. Res. 2020, 116, 1733–1741. [Google Scholar] [CrossRef] [PubMed]
- Chen, L.; Li, X.; Chen, M.; Feng, Y.; Xiong, C. The ACE2 expression in human heart indicates new potential mechanism of heart injury among patients infected with SARS-CoV-2. Cardiovasc. Res. 2020, 116, 1097–1100. [Google Scholar] [CrossRef] [Green Version]
- Hoffmann, M.; Kleine-Weber, H.; Schroeder, S.; Krüger, N.; Herrler, T.; Erichsen, S.; Schiergens, T.S.; Herrler, G.; Wu, N.H.; Nitsche, A.; et al. SARS-CoV-2 cell entry depends on ACE2 and TMPRSS2 and is blocked by a clinically proven protease inhibitor. Cell 2020, 181, 271–280. [Google Scholar] [CrossRef] [PubMed]
- Ou, X.; Liu, Y.; Le, X.; Li, P.; Mi, D.; Ren, L.; Gu, L.; Guo, R.; Chen, T.; Hu, J.; et al. Characterization of spike glycoprotein of SARS-CoV-2 on virus entry and its immune cross-reactivity with SARS-CoV. Nat. Commun. 2020, 11, 1620. [Google Scholar] [CrossRef] [PubMed] [Green Version]
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Huynh, T.V.; Rethi, L.; Lee, T.-W.; Higa, S.; Kao, Y.-H.; Chen, Y.-J. Spike Protein Impairs Mitochondrial Function in Human Cardiomyocytes: Mechanisms Underlying Cardiac Injury in COVID-19. Cells 2023, 12, 877. https://doi.org/10.3390/cells12060877
Huynh TV, Rethi L, Lee T-W, Higa S, Kao Y-H, Chen Y-J. Spike Protein Impairs Mitochondrial Function in Human Cardiomyocytes: Mechanisms Underlying Cardiac Injury in COVID-19. Cells. 2023; 12(6):877. https://doi.org/10.3390/cells12060877
Chicago/Turabian StyleHuynh, Tin Van, Lekha Rethi, Ting-Wei Lee, Satoshi Higa, Yu-Hsun Kao, and Yi-Jen Chen. 2023. "Spike Protein Impairs Mitochondrial Function in Human Cardiomyocytes: Mechanisms Underlying Cardiac Injury in COVID-19" Cells 12, no. 6: 877. https://doi.org/10.3390/cells12060877