The MicroRNAs Profiling Of HIV-infected Patients And The Effect Of MicroRNAs On HIV Replication

HIV could not encode all the molecules required for its life cycle in the host cells because of the limitation of its genome. Inevitability, HIV interacts with many host factors to complete its replication. One kind of these factors are microRNAs, which are19-22nucleotide noncoding RNAs critical for regulation of mRNA translation at the post-transcriptional level. microRNAs bind by a protein complex to the3’untranslated region of mRNA, ultimately resulting in mRNA translational inhibition, degradation, or repression, and play important role in many physiological or pathological processes of human.In the last decade, many studies have shown that changes in expression of endogenous host microRNAs play an important role in HIV infection as well. On one hand, host cellular microRNA profiling will change following HIV infection. On the other hand, certain host cells, such as resting CD4+T cells or monocytes, overexpressed some anti-viral microRNAs to restrict HIV replication or maintain HIV latency. The overexpression of certain cellular microRNAs in vitro has already been shown to reduce HIV-1replication significantly, making the the therapeutic potential of microRNAs in vivo. microRNAs are present in high numbers in a wide range of extracellular media, and researches suggested that the expression levels of microRNAs in the host cells or extracellular media were related to HIV infection progress and prognosis, making the possibility of using microRNAs as novel biomarkers or prognostic factors, independent of more standard assays of immune function and/or markers of immune activation.Clinical application of antiretroviral therapy (ART) can make plasma HIV viremia be suppressed and maintain below the limits of detection for prolonged periods of time, and have changed the natural history of HIV infection in human. In host cells, ART could change the microenvironment in the cells, and the changes of interaction between HIV and host factors was followed. Studies have suggested that HIV infection and antiretroviral therapy have divergent effects on mitochondria in adipose tissue, and could effect the mRNA expression profiling in many other kinds of tissue. Researches on the interaction analysis between microRNA and mRNA and on the effects of ART on microRNA expression profiling are rare. In the present study, we firstly analysed the microRNA profiling between three groups, HIV-infected but ART naive group, HIV-infected with ART group and HIV negative control group, by microarray and qPCR. Then we tested the effects of differentially expressed microRNAs on HIV replication in vitro. Finally, microRNA and mRNA integrated analysis was conducted to explore the mechanism of these effects.In the first section of our research, microRNA expression profiling in the PBMC of the three groups mentioned above was analysed by microarray. Results suggested that overall microRNA expression was restricted in the HIV-infected groups (contain the HIV-infected but ART naive group and the HIV-infected with ART group), and11differentially expressed microRNAs were found. Only one of the11differentially expressed microRNAs, hsa-miR-191-5p, was tested by qPCR in accordance with the microarray data.In the second section of our research, we firstly constructed a vector that could overexpress hsa-miR-191precursor. We then using this vector and hsa-miR-191-5p mimic/inhibitor studied the effect of hsa-miR-191-5p on HIV replication in two in vitro models, the293T/pNL4-3-dE-GFP/pVSV-G pseudotype virus model and the TZM-bl/HIV IIIB HIV infection model. Results suggested that hsa-miR-191-5p could suppress HIV replication in the two models in vitro. And this suppression might be produced by effecting related target molecules of hsa-miR-191-5p.In the third section of our research, we conducted comprehensive search for experimentally confirmed or potential targets of hsa-miR-191-5p that were related to HIV replication, by searching microRNA databases and related articles, by microRNA and mRNA integrated analysis, and by using "HIV-1, Human Protein Interaction Database" to confirm the relationship between the targets and HIV replication. qPCR and reporter gene vector containing3’untranslated region of the potential targets were used to test the effects of hsa-miR-191-5p on the potential targets. Results suggested that CCR1and NUP50might be the targets of hsa-miR-191-5p, and hsa-miR-191-5p might effect HiV replication by suppressing these targets.In summary(1) Differential expression of microRNA in PBMC was observed between three groups, the HiV-infected but ART naive group, the HiV-infected with ART group and the HIV negative control group. Overall, microRNA expression in PBMC was restricted in the HIV-infected groups compared to the HIV negative control.(2) hsa-miR-191-5p in PBMC had differentially expressed between the HIV-infected but ART naive group and the HIV negative control group. And hsa-miR-191-5p has suppressing effects on HIV replication in vitro.(3) CCR1and NUP50might be the targets of hsa-miR-191-5p, and hsa-miR-191-5p might effect HIV replication by suppressing these targets.