利用永生化人间充质干细胞分泌包载抗HIV-1 siRNA的细胞微粒的研究

Acquired immunodeficiency syndrome (AIDS) is mainly caused by infection with human immunodeficiency virus type 1 (HIV-1), which is severely threaten the public health and safety. The hope of HIV-1 therapy was either a sterilising cure (by which in theory all latent HIV DNA is eliminated) or a functional cure (wherein latent HIV persists but viraemia is very low or absent without the use of antiretroviral therapy). Studies of HIV-1 new treatment to reduce AIDS burden or cure are ongoing. Small interfering RNA (siRNA) can inhibit the expression of target gene efficiently and specifically. And cell-derived microvesicles (MVs) are released by many types of cells, exist in almost all types of body fluids, and are important mediators of cell-to-cell communication. The aim of this project is to establish a novel nucleic acid carrier to treat HIV, which is bio-nanoparticles-MVs carried cargoes of specific siRNA——take the siRNA to anti-HIV-1 viral protein R (vpr) gene for example. In the previous study, the MVs could inhibit the expression of HIV-1 vpr, and human mesenchymal stem cells (hMSCs)-derived MVs were obtained. Based on it, MVs produced by immortalized hMSCs long and stably will be established, the appropriate condition for MVs production and purification will be identified, and the effects of MVs on HIV-1 will be tested. The completion of this project will provide a new idea for developing a novel HIV gene therapy and a new clue for siRNA therapy.

艾滋病是一种严重威胁人类健康的重大传染病，以降低疾病负担和治愈疾病为目标的HIV治疗新方法的研究一直在进行。siRNA可特异高效的抑制靶基因的表达；细胞微粒由细胞分泌、体液传输，可在细胞间传递信息。本项目以HIV-1的vpr基因为例，结合siRNA和细胞微粒各自的优势，将siRNA包载到细胞微粒内，构建新型siRNA载体，探索HIV-1治疗新方法。前期工作中发现了包载特异性siRNA的细胞微粒可抑制细胞内HIV-1 vpr基因的表达，并获得了表达siRNA的人间充质干细胞（hMSCs）系。在此基础上，为获得大量的细胞微粒及进一步检测其对HIV-1的抑制作用，本项目拟建立永生化hMSCs长期稳定分泌包载特异性siRNA的细胞微粒的体系，摸索并确认细胞微粒的制备和提纯条件，并在体外检测其对HIV-1的抑制效果。为HIV-1基因治疗提供新思路，同时为siRNA的体内应用提供新方法。

艾滋病是一种严重威胁人类健康的重大传染病，以降低疾病负担和治愈疾病为目标的HIV治疗新方法的研究一直在进行。siRNA可特异高效的抑制靶基因的表达；细胞微粒由细胞分泌、体液传输，可在细胞间传递信息。本项目以HIV-1的vpr基因为例，结合siRNA和细胞微粒各自的优势，将siRNA包载到细胞微粒内，摸索并确认细胞微粒的制备和提纯条件，并在体外检测其对HIV-1的抑制效果。本项目筛选到1株稳定表达siRNA的hMSCs阳性细胞克隆，制备的细胞微粒经过透射电镜、纳米颗粒分析、siRNA、总蛋白含量分析，该微粒对vpr基因的最大表达抑制率为21.06%，对感染后细胞释放HIV-1的最高抑制率为47.87%，同时MTT法检测该剂量的细胞微粒对细胞无明显毒性。这为HIV-1基因治疗提供新思路，同时为siRNA的体内应用提供新方法。

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