Novel tools to study the interplay between peroxisomes and lipid droplets in hepatitis C virus infection

• Autor(a) principal: Carvalho, Ariana dos Santos
• Data de Publicação: 2021
• Tipo de documento: Dissertação
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/10773/33823
• topic: Peroxisomes; Lipid droplets; Hepatitis C virus (HCV); Interorganellar interaction; Lentivirus; Cloning

Hepatitis C represents a serious public health burden, with a high incidence rate worldwide. Infection normally progresses to cirrhosis and liver cancer, and while antiviral treatments are effective, access to diagnosis and treatment is still limited, with no vaccine yet developed. Hepatitis C virus (HCV) takes advantage of several host cell mechanisms through the manipulation of cellular organelles’ functions. HCV specifically modulates lipid droplets (LDs) and critically depends on the cellular lipid metabolism. Peroxisomes are also essential organelles in lipid metabolism and coordinate some of their functions with LDs, though their role as a metabolic organelle in HCV infection is still unknown. With this work, we aimed to create tools to study the effect of HCV infection on peroxisome morphology, cellular distribution, and their interplay with LDs. Moreover, these tools would also allow to understand the importance of this crosstalk for the HCV life cycle. We successfully created plasmids and lentiviruses expressing peroxisome-targeted fluorescent fusion proteins. We have also generated a plasmid expressing an LD-targeted fluorescent protein. Preliminary studies on the influence of HCV core protein overexpression on peroxisome morphology and subcellular localisation have revealed no clear alterations. Further studies are required to improve the LD-targeting fluorescent fusion protein-expressing lentiviruses, which will then allow the generation of stable cell lines with fluorescently stained LDs and peroxisomes. Furthermore, additional morphology and subcellular localisation analyses of peroxisomes throughout infection must be conducted to better understand the role of this organelle in HCV infection. These studies may unravel new roles of peroxisomes in HCV infection which can ultimately lead to the discovery of novel targets for antiviral therapeutics' development.