SARS-CoV-2 infection and replication kinetics in different human cell types: The role of autophagy, cellular metabolism and ACE2 expression

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Bartolomeo, Cynthia Silva ; Lemes, Robertha Mariana Rodrigues ; Morais, Rafael Leite ; Pereria, Gabriela Cruz ; Nunes, Tamires Alves ; Costa, Angelica Jardim ; Maciel, Rui Monteiro de Barros ; Braconi, Carla Torres ; Maricato, Juliana Terzi ; Janini, Luiz Mario Ramos ; Okuda, Liria Hiromi ; Lee, Kil Sun ; Prado, Carla Maximo ; Ureshino, Rodrigo Portes ; Stilhano, Roberta Sessa

• Document type: Journal article
• Source: Life Sciences; v. 308, p. 13-pg., 2022-09-07.

Abstract

Aims: This study evaluated SARS-CoV-2 replication in human cell lines derived from various tissues and inves-tigated molecular mechanisms related to viral infection susceptibility and replication. Main methods: SARS-CoV-2 replication in BEAS-2B and A549 (respiratory tract), HEK-293 T (kidney), HuH7 (liver), SH-SY5Y (brain), MCF7 (breast), Huvec (endothelial) and Caco-2 (intestine) was evaluated by RT-qPCR. Concomitantly, expression levels of ACE2 (Angiotensin Converting Enzyme) and TMPRSS2 were assessed through RT-qPCR and western blot. Proteins related to autophagy and mitochondrial metabolism were moni-tored in uninfected cells to characterize the cellular metabolism of each cell line. The effect of ACE2 over -expression on viral replication in pulmonary cells was also investigated. Key findings: Our data show that HuH7, Caco-2 and MCF7 presented a higher viral load compared to the other cell lines. The increased susceptibility to SARS-CoV-2 infection seems to be associated not only with the differential levels of proteins intrinsically related to energetic metabolism, such as ATP synthase, citrate synthase, COX and NDUFS2 but also with the considerably higher TMPRSS2 mRNA expression. The two least susceptible cell types, BEAS-2B and A549, showed drastically increased SARS-CoV-2 replication capacity when ACE2 was overex-pressed. These modified cell lines are relevant for studying SARS-CoV-2 replication in vitro. Significance: Our data not only reinforce that TMPRSS2 expression and cellular energy metabolism are important molecular mechanisms for SARS-CoV-2 infection and replication, but also indicate that HuH7, MCF7 and Caco-2 are suitable models for mechanistic studies of COVID-19. Moreover, pulmonary cells overexpressing ACE2 can be used to understand mechanisms associated with SARS-CoV-2 replication. (AU)

• FAPESP's process: 20/04709-8 - Evaluation of potential therapeutically compounds for SARS-CoV-2: focus on estrogen-related compounds, autophagy modulators and ACE2
• Grantee: Rodrigo Portes Ureshino
• Support Opportunities: Regular Research Grants
• FAPESP's process: 06/60402-1 - Medular carcinoma of the thyroid: revisiting the clinical, molecular biological, biochemical and biological aspects following findings of molecular genetics
• Grantee: Rui Monteiro de Barros Maciel
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 20/06153-7 - Evaluation of estrogen-related compounds and autophagy modulators seeking for therapeutic targets against SARS-CoV-2
• Grantee: Robertha Mariana Rodrigues Lemes
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 20/13480-4 - Virtual screening and in vitro and in vivo evaluation of nicotinic receptors as a therapeutic target for SARS-CoV-2: focus on interaction with ACE2.
• Grantee: Carla Máximo Prado
• Support Opportunities: Regular Research Grants
• FAPESP's process: 20/08943-5 - Investigation of the hosts' induced elements in response to the immunisation with ChAdOx1 nCOV-19 vaccine in a Phase III Clinical Trial
• Grantee: Luiz Mário Ramos Janini
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 16/20796-2 - Study of estrogen receptors mediated autophagy against tau toxicity in cell and zebrafish models
• Grantee: Rodrigo Portes Ureshino
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 19/10922-9 - Gene and cell therapy via alginate microgels for muscle injuries
• Grantee: Roberta Sessa Stilhano Yamaguchi
• Support Opportunities: Research Grants - Young Investigators Grants