The role of extracellular vesicle-associated MicroRNAs in HIV-associated atherosclerosis

细胞外囊泡相关 MicroRNA 在 HIV 相关动脉粥样硬化中的作用

• 批准号: 10619831
• 负责人: CHUNMING DONG
• 金额: $ 38.38万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2027-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10619831
• 关键词: 3' Untranslated Regions; Acceleration; Acquired Immunodeficiency Syndrome; Adverse effects; Affect; Aging; Apolipoprotein E; Apoptosis; Arterial Fatty Streak; Atherosclerosis; Biological; Body Fluids; Bone Marrow; Bone Marrow Cells; CD4 Lymphocyte Count; Cardiovascular Diseases; Cardiovascular Pathology; Cardiovascular system; Cell Communication; Cell Separation; Cells; Chronic; Data; Development; Distant; Endothelial Cells; Endothelium; Engineering; Equilibrium; Exposure to; Foundations; Genes; Genetic; Goals; Grant; HIV; HIV Infections; HIV Seronegativity; HIV Seropositivity; HMGA2 gene; Human; Impairment; In Vitro; Infection; Inflammation; Inflammatory; Injury; Lipids; Liquid substance; Mediating; Mediator; Messenger RNA; MicroRNAs; Modification; Molecular; Morbidity - disease rate; Mus; Pathway interactions; Persons; Plasma; Prevention strategy; Process; Proliferating; Proteins; Publishing; Quality of life; Regulation; Resistance; Role; Sampling; Small RNA; Smooth Muscle Myocytes; Solid; Testing; Urine; Vascular Endothelial Cell; Vesicle; Viral; Viral Load result; Writing; antagonist; antiretroviral therapy; atherogenesis; candidate validation; cardiovascular disorder risk; cell type; clinical care; endothelial stem cell; exosome; extracellular vesicles; high risk; immune activation; improved; injury and repair; latent HIV reservoir; lonely individuals; mesenchymal stromal cell; migration; mortality; nanosized; novel; overexpression; peripheral blood; premature; reconstitution; senescence; transcriptome; transcriptome sequencing; treatment strategy; vascular injury; virtual

People living with HIV (PLWH) have a significantly higher risk of cardiovascular diseases (CVD) compared to their HIV negative counterparts. However, the underlying mechanisms that drive this increased CVD risk remain elusive. Extracellular vesicles (EVs)/exosomes are released by all cells and widely distributed in plasma, urine, and other biological fluids. EVs contain diverse bioactive molecules, including mRNAs, microRNAs (miRs), proteins, and lipids, which can be transferred to target cells altering their functionality. The cargo of EVs reflects the pathophysiological states of the host cells. It is conceivable that the latent HIV reservoir established during early infection, long-term exposure to antiretroviral therapy (ART), and chronic immune activation/inflammation (referred to as HIV positivity for grant writing purpose) may impact host cells, particularly those in the peripheral blood, altering the release and the cargo of plasma EVs. These EVs in turn may serve as information vehicles for cell-to-cell communication, affecting cells of the arterial wall, including endothelial cells (ECs) and smooth muscle cells (SMCs), as well as endothelial progenitor cells (EPCs) in the bone marrow, promoting atherogenesis. To test this hypothesis, we fractionated plasma from PLWH on ART with undetectable viral load into plasma EVs (HIVposEVs) and plasma-depleted of EVs (HIVplasmadepEVs) and injected these fractions into atherogenic apoE-/- mice. Remarkably, HIVposEVs, but not HIVplasmadepEVs, increased atherosclerosis plaque burden compared to EVs isolated from HIV negative subjects (HIVnegEVs) and PBS. These changes were accompanied by increased senescence and apoptosis of ECs and SMCs in the arterial wall and impaired viability and functionality of bone marrow EPCs. These exciting data support the hypothesis that circulating EVs from PLWH reconstitute the effects of HIV positivity (without active HIV infection) on atherosclerosis. RNA-seq in HIVposEVs and HIVnegEVs revealed differential miR profiles. The target genes of EV-miRs overrepresented in PLWH are enriched in cardiovascular pathology pathways. We have engineered tailored EVs (TEVs) that are loaded with the antagomirs for let-7b-5p and miR103a-2, two candidate EV-miRs overrepresented in HIVposEVs by transducing mesenchymal stromal cells. These TEVs counteracted the effects of HIVposEVs on the accelerated senescence of EPCs, whereas let-7b- 5p overexpressing EVs recapitulated, in part, the effects of HIVposEVs. Furthermore, EPCs overexpressing HMGA2 (a top let-7b-5p target gene) lacking the 3'UTR (resistant to miR regulation) showed protection similar to miRZip-let-7b TEVs against HIVposEVs-induced senescence. In this proposal, we will determine if TEVs engineered to modulate key HIVposEVs-associated miRs counteract the effects of the chronic inflammation and immune activation, ART, and latent HIV reservoir associated with HIV positivity on atherosclerosis development. In addition, we will further dissect the mechanisms underlying the adverse effect of HIV positivity on the cardiovascular system by examining the effects of HIVposEVs and/or TEVs on the functionality and transcriptome of ECs, SMCs and EPCs. We will identify putative miR-mRNA pathways. Importantly, we will validate our mouse findings in EPC samples from PLWH using TEVs, direct miR antagonism, and target gene overexpression approaches.

艾滋病毒（PLWH）患者患心血管疾病（CVD）的风险明显更高 与他们的艾滋病毒负面同行。但是，驱动这种增加CVD风险的基本机制 保持难以捉摸。细胞外囊泡（EV）/外泌体均由所有细胞释放，并广泛分布在 血浆，尿液和其他生物液。电动汽车包含多种生物活性分子，包括mRNA， microRNA（miR），蛋白质和脂质，可以转移到靶细胞改变其功能的目标。 电动汽车的货物反映了宿主细胞的病理生理状态。可以想象，潜伏的艾滋病毒 早期感染期间建立的储层，长期暴露于抗逆转录病毒疗法（ART）和慢性 免疫激活/炎症（称为授予写作目的的艾滋病毒阳性）可能会影响主机 细胞，特别是外周血中的细胞，改变了血浆EV的释放和货物。这些电动汽车 反过 包括内皮细胞（EC）和平滑肌细胞（SMC）以及内皮祖细胞细胞 （EPC）在骨髓中，促进动脉粥样硬化。为了检验该假设，我们从 在ART上具有无法检测到的病毒载荷到血浆EV（Hivposevs）的ART上的PLWH，并耗尽EV的等离子体 （Hivplasmadepevs）并将这些馏分注入动脉粥样硬化的APOE - / - 小鼠。值得注意的是，Hivposevs，但没有 Hivplasmadepevs，与HIV阴性分离的EV相比，动脉粥样硬化斑块负担增加 受试者（Hivn​​egevs）和PBS。这些变化伴随着衰老和凋亡的增加 动脉壁中的EC和SMC的骨髓骨髓EPC的活力和功能受损。这些 令人兴奋的数据支持以下假设，即PLWH循环电动汽车重建了HIV阳性的影响 （没有活性HIV感染）动脉粥样硬化。 HIVPOSEV和HIVNEGEVS中的RNA-Seq揭示了差异miR 概况。 PLWH中EV-MIRS的靶基因富含心血管病理学 途径。我们已经设计了量身定制的电动汽车（TEV），这些电动汽车装有let-7b-5p的Antagomirs和 miR103a-2，两个候选EV-MIRS通过转导间充质基质细胞在HIVPOSES中的代表性过高。 这些TEV抵消了Hivposev对EPC加速衰老的影响，而Let-7b- 5p过表达的EV概括了HIVPOSEV的作用。此外，EPC过表达 缺乏3'UTR（对MIR调节的抗MIR调节）的HMGA2（最高的Let-7b-5p靶基因）显示保护 类似于针对HIVPOSEVS诱导的衰老的mirzip-let-7b TEV。在此提案中，我们将确定是否 TEVS设计为调节密钥HIVPOSEVS相关的miRs抵消了慢性的影响 与HIV阳性相关的炎症和免疫激活，ART和潜在的HIV储藏 动脉粥样硬化的发展。此外，我们将进一步剖析不利的机制 HIV阳性对心血管系统的影响，通过检查HIVPOSEV和/或TEV对 EC，SMC和EPC的功能和转录组。我们将确定推定的miR-mRNA途径。 重要的是，我们将使用TEV验证PLWH中的EPC样品中的鼠标发现，直接mir 对抗和靶基因过表达方法。