small RNA sequencing from plasma samples of HIV/HCV coinfected individuals
High-throughput sequencing of the miRNAs present in plasma of people with HIV (PWH) who have clearance HCV after completion of antiviral therapy. This study allowed us to identify and functionally characterize human miRNAs associated with the development of long-term hepatic and non-hepatic events. Samples were collected at three points: baseline (before HCV therapy), and one and five years after completion of HCV therapy. All individuals were on stable combination antiretroviral therapy (cART) for at least six months and had an undetectable plasma HIV viral load (<50 copies/mL). PWH with acute hepatitis C, hepatitis B virus (HBV), or a prior history of liver decompensation were excluded from the study. Plasma fraction was obtained using Ficoll-Paque density gradient centrifugation and, subsequently, stored at the Spanish HIV HGM Biobank at -80ºC. Total RNA, including small RNAs, was isolated from 600µl of plasma with the miRNeasy Serum Plasma Advanced kit (Qiagen). RNA quality and quantity were evaluated by the Agilent Bioanalyzer HS DNA Assay (ref. 5067-4626) and KAPA Library Quantification Kit KK4835 (ref. 07960204001, Roche).