miRNAs involved in HIV infection. Effects of cocaine use on course of infection

MORANDO N; GUZMAN L; RABINOVICH D; PANDO MA

Bernal

Congreso; II Reunión Argentina de ARNs no codificantes; 2018

Universidad Nacional de Quilmes

Objective: The main goal of the proposed study is to evaluate the effect of exposure to substances of abuse, particularly adulterated cocaine (caffeine being the most common adulterant used in Argentina), on the expression of miRNAs involved in HIV infection. Previous studies have shown that the miRNA profile is altered by the consumption of drugs of abuse. Furthermore, it has also been proven that drugs of abuse modulate the secretion of extracellular vesicles (EV). However, the role of drugs of abuse in the secretion of miRNAs associated with EV has not been explored. Consequently, we propose to assess whether there is a difference between the miRNA profiles found intracellularly, within VE or extracellularly in carrier proteins.Experimental design: we propose to evaluate the miRNA profile in peripheral blood samples from individuals who report using/not using cocaine, with/without HIV infection (20 individuals in each group). Drug exposure will be determined in peripheral blood by a diagnostic test. Samples will be processed in order to obtain serum, plasma, whole blood and PBMC and stored at -80°C. VE separation in plasma will be performed via size-exclusion chromatography. This technique has been successfully implemented at the INBIRS (Dr. M. Ostrowski). miRNA extraction from each sample fraction will be carried out using TRIzol LS, TriPure or a commercial kit (mirVana, Ambion, USA). Determination of miRNA profiles will be carried out using the miRNA-seq technique. Library construction and RNA-seq will be carried out by the company Macrogen. miRNAs with a significantly differential expression between study groups will be validated by qRT-PCR. Data analysis will be carried out in collaboration with Dr. M. Rosenzvit (IMPaM, UBA-CONICET).