Engineering a 3D human neurovascular unit for investigating and countering neuroinflammatory mechanisms of HIV

设计 3D 人类神经血管单元来研究和对抗 HIV 的神经炎症机制

• 批准号: 9502663
• 负责人: SURYARAM GUMMULURU
• 金额: $ 26.54万
• 依托单位: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9502663
• 关键词: AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Address; Anti-Retroviral Agents; Antiviral Agents; Apical; Astrocytes; Automobile Driving; Biological; Biological Models; Blood - brain barrier anatomy; Brain; CCL2 gene; CXCL10 gene; Cardiovascular Diseases; Cell Survival; Cells; Chronic; Clinical; Comorbidity; Development; Disease; Drug Targeting; Engineering; Exposure to; Functional disorder; Gene Expression; Genetic Transcription; HIV; HIV Envelope Protein gp120; HIV Infections; HIV-1; HIV-associated neurocognitive disorder; Health; Human; Immune Cell Activation; Immunologic Surveillance; Immunologics; In Vitro; Incidence; Individual; Infection; Inflammation; Inflammatory; Inflammatory Response; Innate Immune System; Interferons; Interleukin-1 beta; Investigation; Macrophage Activation; Malignant Neoplasms; Microfluidics; Microglia; Modeling; Molecular; Morbidity - disease rate; Myeloid Cells; Neurocognitive Deficit; Neurologic; Neurologic Dysfunctions; Neurons; Neuropathogenesis; Older Population; Osteoporosis; Pathogenesis; Patients; Performance; Pericytes; Permeability; Pharmaceutical Preparations; Phase; Process; Production; Proviruses; RNA; Research; Research Priority; Residual state; Risk; Role; SIV; Secondary to; Signal Transduction; TNF gene; Testing; Therapeutic Intervention; Tissues; United States National Institutes of Health; Viral; Viral Genes; Viral Load result; Viral Proteins; Viral reservoir; Virus; Virus Diseases; Virus Replication; Work; age related; antiretroviral therapy; base; brain endothelial cell; cytokine; fluid flow; high risk; immune activation; improved; in vitro Model; in vivo; in vivo Model; induced pluripotent stem cell; inflammatory marker; insight; macrophage; monocyte; mortality; neurocognitive disorder; neuroinflammation; neurotoxic; neurovascular unit; nonhuman primate; novel; novel strategies; prevent; response; small molecule inhibitor; success; therapeutic development; three-dimensional modeling; viral RNA

ABSTRACT Despite effective combination anti-retroviral therapy (cART), HIV-infected individuals remain at an unusually high risk of morbidity and mortality from HIV-associated non-AIDS clinical conditions, such as cardiovascular disease, osteoporosis and neurocognitive decline. HIV-1 reservoir in the CNS is established in the primary phase of virus infection and can result in the development of HIV-1 associated neurocognitive disorders (HAND). While cART can dramatically reduce virus levels to undetectable levels in the CSF and improves many of the neurocognitive disorders, HIV RNA has been observed in the CSF of patients on cART that might contribute to local inflammatory processes. The excess risk for HAND is attributed to the residual immune activation that persists despite cART. Most studies point to tissue-resident cells of the myeloid lineage, perivascular macrophages and microglia as the predominant infected cells in the CNS, though mechanisms that account for chronic immune activation are varied, and include expression of neurotoxic viral proteins, gp120 and Tat, or excess production of pro-inflammatory cytokines, such as TNFα, IL-1β, and MCP-1 by macrophages and microglia in response to residual viral replication. Interestingly, our preliminary findings suggest that de novo expression of HIV unspliced RNA alone from proviruses in productively infected macrophages (in the absence of viral protein production) can induce production of type I IFN responses and pro-inflammatory cytokines, IP-10 and MCP-1, suggesting that expression of viral RNA can perpetuate HAND-inducing inflammatory cycle. To better understand the mechanistic underpinnings of pathophysiology of HAND, we propose two specific aims. In aim 1, we will develop a model of neurovascular unit (NVU) in a microfluidic platform that will consist solely of primary human neuronal cells (pericytes, astrocytes, microglia and neurons) separated from the apical fluid flow by a human brain endothelial cell derived blood-brain-barrier (BBB). Using this novel in vitro primary human cell-derived model system, we will determine the impact of HIV-macrophage induced pro-inflammatory responses on BBB permeability and neuronal cell viability. In aim 2, we will determine the consequences of antiretrovirals that target viral gene expression alone as novel adjunct therapy for suppressing over-exuberant pro-inflammatory responses in the CNS. We predict that these studies will lead to the identification of the biological mechanisms that drive HAND despite effective cART. Furthermore, such insight will be critically important for development of effective strategies to decrease or reverse the persistent immune activation driving disease pathogenesis in the growing population of older individuals living with HIV.

摘要 尽管联合抗逆转录病毒疗法（cART）有效，艾滋病毒感染者仍然处于异常高的水平。 与艾滋病毒相关的非艾滋病临床病症，如心血管疾病， 骨质疏松症和神经认知能力下降。HIV-1在中枢神经系统的储库是在病毒的初级阶段建立的 感染，并可能导致HIV-1相关的神经认知障碍（HAND）的发展。虽然cART 可以显着降低病毒水平，在CSF中检测不到的水平，并改善许多神经认知功能， 在这些疾病中，在接受cART的患者的CSF中观察到HIV RNA， 炎症过程。HAND的额外风险归因于持续存在的残余免疫激活 尽管卡特。大多数研究指出，髓系的组织驻留细胞、血管周围巨噬细胞和 小胶质细胞是CNS中主要的感染细胞，尽管慢性免疫应答的机制是由小胶质细胞引起的。 激活是多种多样的，包括神经毒性病毒蛋白、gp 120和达特的表达，或过量产生 巨噬细胞和小胶质细胞对促炎细胞因子如TNFα、IL-1β和MCP-1的反应 残余病毒复制。有趣的是，我们的初步研究结果表明，未剪接的艾滋病毒的从头表达， 在生产性感染的巨噬细胞中（在没有病毒蛋白生产的情况下）， 诱导产生I型IFN应答和促炎细胞因子IP-10和MCP-1，表明 病毒RNA的表达可以使HAND诱导的炎症循环持续下去。更好地了解 HAND病理生理学的机械基础，我们提出了两个具体的目标。在目标1中，我们将开发 微流体平台中的神经血管单元（NVU）模型，其将仅由原代人类神经元组成， 细胞（周细胞、星形胶质细胞、小胶质细胞和神经元）从人脑顶端液流中分离出来 内皮细胞源性血脑屏障（BBB）。使用这种新的体外原代人类细胞衍生模型 系统，我们将确定HIV-巨噬细胞诱导的促炎反应对BBB的影响 渗透性和神经元细胞活力。在目标2中，我们将确定抗逆转录病毒药物针对 单独的病毒基因表达作为抑制过度旺盛的促炎性反应的新辅助疗法 CNS的反应。我们预测，这些研究将导致确定的生物机制， 尽管有有效的CART，但仍驱动HAND。此外，这种洞察力对发展 有效的策略，以减少或逆转持续的免疫激活驱动的疾病发病机制， 越来越多的老年人感染艾滋病毒。