itochondrial DNA and TLR9 activation contribute to SARS-CoV-2-induced endothelial cell damag

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Costa, Tiago J. ^{[1, 2]} ; Potje, Simone R. ^{[1, 3]} ; Fraga-Silva, Thais F. C. ^[4] ; Da Silva-Neto, Julio A. ^[1] ; Barros, Paula R. ^[1] ; Rodrigues, Daniel ^[1] ; Machado, Mirele R. ^[1] ; Martins, Ronaldo B. ^[5] ; Santos-Eichler, Rosangela A. ^[6] ; Benatti, Maira N. ^[7] ; de Sa, Keyla S. G. ^[8] ; Almado, Carlos Eduardo L. ^[9] ; Castro, Italo A. ^[5] ; Pontelli, Marjorie C. ^[5] ; La Serra, Leonardo ^[5] ; Carneiro, Fernando S. ^[1] ; Becari, Christiane ^[10] ; Louzada-Junior, Paulo ^[7] ; Oliveira, Rene D. R. ^[7] ; Zamboni, Dario S. ^[8] ; Arruda, Eurico ^[5] ; Auxiliadora-Martins, Maria ^[10] ; Giachini, Fernanda R. C. ^[9] ; Bonato, Vania L. D. ^[4] ; Zachara, Natasha E. ^[2] ; Bomfim, Gisele F. ^[11] ; Tostes, Rita C. ^[1]

Afiliação do(s) autor(es):Mostrar menos -

^[1] Univ Sao Paulo, Ribeirao Preto Med Sch, Dept Pharmacol, Ribeirao Preto, SP - Brazil

^[2] Johns Hopkins Univ, Sch Med, Dept Biol Chem, Baltimore, MD 21218 - USA

^[3] Minas Gerais State Univ UEMG, Belo Horizonte, MG - Brazil

^[4] Univ Sao Paulo, Ribeirao Preto Med Sch, Dept Biochem & Immunol, Ribeirao Preto, SP - Brazil

^[5] Univ Sao Paulo, Ribeirao Preto Med Sch, Virol Res Ctr, Ribeirao Preto, SP - Brazil

^[6] Univ Sao Paulo, Inst Biomed Sci, Dept Pharmacol, Ribeirao Preto, SP - Brazil

^[7] Univ Sao Paulo, Dept Clin Med, Div Internal Med, Ribeirao Preto Med Sch, Ribeirao Preto, SP - Brazil

^[8] Univ Sao Paulo, Dept Cell & Mol Biol, Ribeirao Preto Med Sch, Ribeirao Preto, SP - Brazil

^[9] Fed Univ Mato Grosso UFMT, Inst Biol & Hlth Sci, Cuiaba - Brazil

^[10] Univ Sao Paulo, Dept Surg & Anat, Ribeirao Preto Med Sch, Ribeirao Preto, SP - Brazil

^[11] Fed Univ Mato Grosso UFMT, Inst Hlth Sci, Cuiaba - Brazil

• Tipo de documento: Artigo Científico
• Fonte: VASCULAR PHARMACOLOGY; v. 142, FEB 2022.
• Citações Web of Science: 1

Resumo

Background and purpose: Mitochondria play a central role in the host response to viral infection and immunity, being key to antiviral signaling and exacerbating inflammatory processes. Mitochondria and Toll-like receptor (TLR) have been suggested as potential targets in SARS-CoV-2 infection. However, the involvement of TLR9 in SARS-Cov-2-induced endothelial dysfunction and potential contribution to cardiovascular complications in COVID-19 have not been demonstrated. This study determined whether infection of endothelial cells by SARSCoV-2 affects mitochondrial function and induces mitochondrial DNA (mtDNA) release. We also questioned whether TLR9 signaling mediates the inflammatory responses induced by SARS-CoV-2 in endothelial cells. Experimental approach: Human umbilical vein endothelial cells (HUVECs) were infected by SARS-CoV-2 and immunofluorescence was used to confirm the infection. Mitochondrial function was analyzed by specific probes and mtDNA levels by real-time polymerase chain reaction (RT-PCR). Inflammatory markers were measured by ELISA, protein expression by western blot, intracellular calcium (Ca2+) by FLUOR-4, and vascular reactivity with a myography. Key results: SARS-CoV-2 infected HUVECs, which express ACE2 and TMPRSS2 proteins, and promoted mitochondrial dysfunction, i.e. it increased mitochondria-derived superoxide anion, mitochondrial membrane potential, and mtDNA release, leading to activation of TLR9 and NF-kB, and release of cytokines. SARS-CoV-2 also decreased nitric oxide synthase (eNOS) expression and inhibited Ca2+ responses in endothelial cells. TLR9 blockade reduced SARS-CoV-2-induced IL-6 release and prevented decreased eNOS expression. mtDNA increased vascular reactivity to endothelin-1 (ET-1) in arteries from wild type, but not TLR9 knockout mice. These events were recapitulated in serum samples from COVID-19 patients, that exhibited increased levels of mtDNA compared to sex-and age-matched healthy subjects and patients with comorbidities. Conclusion and applications: SARS-CoV-2 infection impairs mitochondrial function and activates TLR9 signaling in endothelial cells. TLR9 triggers inflammatory responses that lead to endothelial cell dysfunction, potentially contributing to the severity of symptoms in COVID-19. Targeting mitochondrial metabolic pathways may help to define novel therapeutic strategies for COVID-19. (AU)

• Processo FAPESP: 13/08216-2 - CPDI - Centro de Pesquisa em Doenças Inflamatórias
• Beneficiário: Fernando de Queiroz Cunha
• Modalidade de apoio: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs
• Processo FAPESP: 16/21239-0 - Estrutura da caveola e sua modulação pelo glicocálice e o processo de mecanotransdução na hipertensão essencial
• Beneficiário: Simone Regina Potje
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 17/25116-2 - Papel da O-GlcNAcilação (O-GlcNAc) na expressão e função do receptor de estrogênio alfa clássico (ERa66kDa) e do splice variant do receptor de estrogênio alfa (ERa36kDa) em artéria carótida comum de camundongos envelhecidos
• Beneficiário: Tiago Januário da Costa
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado