Dados do Trabalho

Título

DEVELOPMENT OF LENTIVIRAL VECTORS FOR INHIBITION OF HEPATITIS B VIRUS, VIA SMALL INTERFERING RNA

Introdução

It is estimated that chronic hepatitis B virus (HBV) infection accounts for one million deaths/year due to cirrhosis and liver cancer. Despite available treatment options, complete cure of the disease is not achieved primarily due to the persistence of HBV DNA in the host nucleus, presented as a covalently closed circular DNA (cccDNA) or integrated into the host genome, enabling viral reactivation. In this context, Interfering RNA (RNAi) emerges as a promising alternative for chronic hepatitis treatment by enabling post-transcriptional gene silencing.

Objetivo (s)

This study aimed to design and evaluate the efficacy of RNAi lentiviral vector targeting key HBV proteins (HBsAg, HBcAg, HBeAg) and pre-genomic RNA (pgRNA) for silencing.

Material e Métodos

The silencing vector candidates were designed to target overlapped Open Reading Frames (ORFs), allowing simultaneous silencing of multiple viral proteins and pgRNA with a single RNAi. Three silencing vector candidates were constructed and tested in silico to prevent off-target effects. Stability and secondary structures have also been evaluated. Huh7 cells were transfected with 1 µg of purified HBV genotype A circular monomers, followed by infection with the first lentiviral candidate (siHBV-1), targeting S/Pol genes of HBV (108 TU/mL), 3 days later. Quantification of HBV proteins using chemiluminescence and HBV DNA using quantitative PCR (qPCR) was conducted throughout the post-transfection period.

Resultados e Conclusão

Results demonstrated effective silencing of HBsAg expression in cells infected with siHBV-1, with undetectable levels observed from the third day post-infection compared to untreated controls (p

Palavras Chave

HBV; RNAi; gene therapy; lentiviral vector