648. Non-Coding RNAs as Clinically Relevant Host Factors and Target Molecules in HIV-1 Infection
Authors: Kathleen Boerner, Elisabeth Fischer, Paul Schnitzler, Puri Fortes, Hans-Georg Kräusslich, Dirk Grimm
Journal: Molecular Therapy (2015)

Over the past decade, non-coding RNAs comprising miRNAs and lncRNAs (long non-coding RNAs) have gained tremendous attention due to their fundamental biological role as gatekeepers of cellular protein expression. Concurrently, they have become attractive from a clinical standpoint based on their frequent dysregulation in human diseases including cancers and viral infections. Particularly interesting in this respect is accumulating evidence that miRNAs are secreted from somatic cells into various easily accessible body fluids where they likewise form signatures that may describe physiological or pathological states of the host. Ideally, a thorough characterization of non-coding RNAs that are dysregulated in cells and extra-cellular fluids will not only improve our knowledge of human pathologies, but also yield additional biomarkers for disease monitoring, novel targets for clinical intervention and new avenues for creation of therapeutic vectors that are sensitive to these perturbations. Towards these aims, we focused on a major pathogen, the human immunodeficiency virus HIV-1, and asked how HIV-1 infection would dysregulate intra- and extra-cellular non-coding RNA profiles in the blood of adult humans. We thus profiled a subset of human miRNAs in HIV-1-infected patients as well as healthy age- and gender-matched controls, using a combination of microarrays and qRT-PCR for miRNA detection and quantification. In more detail, we assembled a cohort of 20 HIV-1-infected subjects with a viral load of at least 103 HIV genomes per ml, and then extracted total RNA from whole blood and cellfree plasma for miRNA profiling. The resulting comprehensive signatures of HIV-associated intra- or extra-cellular miRNAs were next compared amongst each other, as well as to those from the control cohort of HIV-1-negative adults. Notably, we identified 11 miRNAs that were specifically and significantly upregulated in the blood cells of HIV-1-infected subjects versus controls, accompanied by wide-spread increases of further miRNAs in the plasma of this cohort. Interestingly, we moreover measured intra-cellular HIV-1-dependent elevations of ISR2, a lncRNA that we recently also found to be induced by interferons as well as other pathogens including hepatitis C and influenza virus (Carnero et al., Front Immunol 2014;5:548). As a whole, our data validate that HIV-1 infection triggers a massive dysregulation of miRNA and lncRNA expression in blood cells of affected patients and a concurrent perturbation of cellfree miRNA signatures. While some of these candidates may indicate a generalized response to infection, our hope is that these non-coding RNA fingerprints could ultimately provide new clinically relevant and specific biomarkers and targets for HIV-1 diagnosis, prognosis or therapy. In addition, the observed discrepancies between intra- and extra-cellular miRNA profiles yield numerous starting points for further investigation into fundamental miRNA biology in humans.