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The effect of SARS-CoV-2 structural proteins on the expression of cytokines and APOBEC genes in human cell lines
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A pandemia global causada pelo Coronavírus 2 da Síndrome Respiratória Aguda Grave (SARS-CoV-2) despertou uma necessidade urgente de compreender as interações do vírus com o hospedeiro humano a nível molecular.
Os efeitos nocivos da Doença por Coronavírus 2019 (COVID-19) são cada vez mais atribuídos a danos imunopatológicos causados pelo aumento das citocinas pró-inflamatórias, estabelecendo uma ligação entre a tempestade de citocinas e as complicações graves da doença.
As proteínas APOBEC pertencem a uma família de deaminases de citidina responsáveis pela edição da sequência de ADN e ARN, desempenhando papéis fundamentais numa vasta gama de processos biológicos, incluindo respostas imunitárias, propriedades antivirais e mutações genéticas.
Neste trabalho, investigamos e reportamos dados sobre o efeito das proteínas estruturais do SARS-CoV-2 - Envelope (E), Spike (S), Nucleocápside (N) e Membrana (M) - na expressão das citocinas Interleucina 8 (IL-8) e Fator de Necrose Tumoral-alfa (TNF-α), e das proteínas APOBEC3s - APOBEC3B, APOBEC3F e APOBEC3G - nas linhas celulares humanas, Huh-7 e A549.
Apesar de existir evidência científica acerca dos efeitos do SARS-CoV-2 na cascata inflamatória, a literatura atual referente ao impacto do SARS-CoV-2 na expressão das APOBEC é escassa. Desta forma, focámo-nos em conceber um protocolo para quantificarmos o efeito das proteínas estruturais do SARS-CoV-2 na expressão de citocinas inflamatórias e APOBECs acima mencionadas, através do método de qPCR.
Os nossos resultados revelam uma relação complexa entre as proteínas estruturais do SARS-CoV-2 e a resposta imunitária do hospedeiro, visto que certas proteínas estruturais virais modulam a expressão de citocinas, contribuindo potencialmente para as respostas imunitárias desreguladas observadas nos doentes com COVID-19. Além disso, os resultados obtidos apontam para possíveis interações entre as proteínas virais e as APOBEC.
Este estudo contribui para compreensão das interações vírus-hospedeiro no contexto da infeção por SARS-CoV-2 e fornece alguma informação sobre potenciais alvos terapêuticos com vista a atenuar as consequências imunopatológicas da doença.
The global pandemic caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) triggered an urgent need to understand the virus's interactions with the human host at the molecular level. The detrimental effects of Coronavirus disease 19 (COVID-19) are increasingly attributed to immunopathological damage caused by increased pro-inflammatory cytokines, establishing a link between the cytokine storm and severe complications of the disease. The Apolipoprotein B mRNA editing catalytic polypeptide-like (APOBEC) proteins belong to a family of cytidine deaminases responsible for DNA and RNA sequence editing, playing pivotal roles in a wide range of biological processes, including immune responses, antiviral properties, and genetic mutations. In this work, we investigate and report data of the effect of SARS-CoV-2 structural proteins - Envelope (E), Spike (S), Nucleocapsid (N) and Membrane (M) - on the expression of cytokines Interleukin 8 (IL-8) and Tumor Necrosis Factor-alpha (TNF-α), and APOBEC3s proteins (APOBEC3B, APOBEC3F and APOBEC3G) genes in Huh-7 and A549 human cell lines. While there is a lot of scientific evidence about the effects of SARS-CoV-2 on the inflammatory cascade, the current literature regarding the impact of SARS-CoV-2 on APOBEC expression is scarce. Therefore, we focused on designing a protocol to quantify the effect of SARS-CoV-2 structural proteins on the expression of the aforementioned inflammatory cytokines and APOBECs using qPCR. Our findings reveal a complex relationship between SARS-CoV-2 structural proteins and the host immune response, as certain viral structural proteins modulate cytokines expression, potentially contributing to the dysregulated immune responses seen in COVID-19 patients. Additionally, our research uncovered interactions between viral proteins and APOBEC genes. This study contributes to a better understanding of the host-virus interactions in the context of SARS-CoV-2 infection and provides some insights into potential therapeutic targets for mitigating the immunopathological consequences of the disease.
The global pandemic caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) triggered an urgent need to understand the virus's interactions with the human host at the molecular level. The detrimental effects of Coronavirus disease 19 (COVID-19) are increasingly attributed to immunopathological damage caused by increased pro-inflammatory cytokines, establishing a link between the cytokine storm and severe complications of the disease. The Apolipoprotein B mRNA editing catalytic polypeptide-like (APOBEC) proteins belong to a family of cytidine deaminases responsible for DNA and RNA sequence editing, playing pivotal roles in a wide range of biological processes, including immune responses, antiviral properties, and genetic mutations. In this work, we investigate and report data of the effect of SARS-CoV-2 structural proteins - Envelope (E), Spike (S), Nucleocapsid (N) and Membrane (M) - on the expression of cytokines Interleukin 8 (IL-8) and Tumor Necrosis Factor-alpha (TNF-α), and APOBEC3s proteins (APOBEC3B, APOBEC3F and APOBEC3G) genes in Huh-7 and A549 human cell lines. While there is a lot of scientific evidence about the effects of SARS-CoV-2 on the inflammatory cascade, the current literature regarding the impact of SARS-CoV-2 on APOBEC expression is scarce. Therefore, we focused on designing a protocol to quantify the effect of SARS-CoV-2 structural proteins on the expression of the aforementioned inflammatory cytokines and APOBECs using qPCR. Our findings reveal a complex relationship between SARS-CoV-2 structural proteins and the host immune response, as certain viral structural proteins modulate cytokines expression, potentially contributing to the dysregulated immune responses seen in COVID-19 patients. Additionally, our research uncovered interactions between viral proteins and APOBEC genes. This study contributes to a better understanding of the host-virus interactions in the context of SARS-CoV-2 infection and provides some insights into potential therapeutic targets for mitigating the immunopathological consequences of the disease.
Descrição
Trabalho Final de Mestrado Integrado, Ciências Farmacêuticas, 2023, Universidade de Lisboa, Faculdade de Farmácia.
Palavras-chave
Sars-CoV-2 APOBEC3B APOBEC3F APOBEC3G IL-8 TNF-alpha Mestrado Integrado - 2023