Alcohol and drug abuse are key co-morbidities of HIV-1 infection. The advent of combination anti-retroviral therapy (cART) has greatly improved the life expectancy of HIV-1-infected individuals, and there has been shift in research from the study of the deleterious effects of addictive substances on the progression of HIV infection, to examination of whether the viral infection itself can lead to addiction. Using the HIV-1Tg rat model, we showed that the intensified addictive behavior in HIV-1Tg rats correlates with the presence of viral proteins and that chronic exposure to alcohol further enhances the novelty seeking behaviors associated with susceptibility to substance abuse. We recently employed CRISPR/Cas9 gene editing technology to target and eradicate the HIV-1 genome from the chromosomes of infected cells. By targeting LTR sequences flanking the integrated HIV-1 pro-virus, we were able to completely excise the full length viral genome from latently infected cell lines. Similar results were achieved on in vitro infected PBMCs and CD4+ T cells derived from healthy donors as well as ex vivo cultured cells from HIV-1 positive patients. We combined multiplex gRNAs directed to different regions of the viral genome with AAV9-based delivery to show, for the first time, successful excision of viral sequences in a variety of tissues from HIV-1Tg mice and rats. The removal of the HIV-1 genome caused a significant drop in viral RNA expression in many tissues, including the blood, brain, and liver. Based on those data, we propose to utilize our gene editing therapy to correct the HIV-1- and alcohol-induced molecular and behavioral malfunctions in the HIV-1Tg rat model. The findings from these molecular and behavioral pre- clinical studies will set the stage for the development of gene therapies designed specifically to treat alcohol- addicted HIV-1 patients on cART.
Alcohol abuse is one of the most common co-morbidities in HIV-1 infection. Using the HIV-1 transgenic rat model, the goal of this exploratory project is to demonstrate the use gene editing technology to reverse alcohol-induced metabolic and behavioral malfunctions in HIV-1 patients on cART.