Functional Characterisation of Host MicroRNA Regulation of Hepatitis C Virus Replication in Nigerian Genotype 1 and Genotype 2 Infected Hepatocytes

MicroRNAs (miRNAs) are endogenous small non-coding RNAs that regulate HCV replication through complex interactions with viral RNA elements and host antiviral pathways. miR-122, liver-enriched and known to enhance HCV replication, is the most characterised, yet the global miRNA-HCV regulatory network in the context of Nigerian genotype 1 and genotype 2 HCV strains is uncharacterised. This study functionally characterises host miRNA regulation of HCV replication in hepatocytes derived from genotype 1a and genotype 2a HCV-infected Nigerian patients. Primary hepatocytes and Huh7.5 cell lines will be infected with patient-derived HCV isolates from Lagos, Abuja, and Port Harcourt. Next-generation small RNA sequencing will profile the miRNA landscape of infected versus uninfected hepatocytes at 24, 72, and 120 hours post-infection. Bioinformatics prediction tools (miRanda, TargetScan) will identify HCV genome-targeting miRNA candidates from differentially expressed miRNAs. Functional validation using miRNA mimics and antagomirs will quantify the impact of candidate miRNAs on HCV RNA replication, NS5A protein expression, and infectious virus production. Cross-genotype comparisons will identify miRNA interactions specific to Nigerian genotype 2 strains, which are genetically distinct from Asian and European counterparts. This study makes an original molecular virology contribution by defining the miRNA regulatory landscape of HCV genotypes prevalent in Nigeria, potentially identifying novel RNA-based antiviral targets applicable to the developing West African HCV therapeutic pipeline. Keywords: Hepatitis C virus, miRNA regulation, genotype 2, HCV replication, Nigeria