The hCNS-HIV/ARV assay system, a platform utilizing human iPSC-neurons/-microglia/-astrocytes to test HIV/AIDS therapeutics for neurotoxicity related to HIV-associated cognitive disroder (HAND)

hCNS-HIV/ARV 检测系统是一个利用人类 iPSC-神经元/-小胶质细胞/-星形胶质细胞来测试 HIV/AIDS 疗法与 HIV 相关认知障碍 (HAND) 相关神经毒性的平台

• 批准号: 10259617
• 负责人: PATRICK M MCDONOUGH
• 金额: $ 97.52万
• 依托单位: VALA SCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-10 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10259617
• 关键词: AIDS dementia; AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Adult; Affect; Aging; Alzheimer' s Disease; Animals; Anti-Retroviral Agents; Astrocytes; Autophagocytosis; Biological Assay; Bipolar Disorder; Birth; Brain; Calcium; Cells; Child; Coculture Techniques; Cognition; Cognition Disorders; Cognitive; Collaborations; Congenital Abnormality; Data; Dementia; Development; Developmental Delay Disorders; Diagnosis; Embryo; Epigenetic Process; Exhibits; Goals; HIV; HIV Infections; HIV antiretroviral; Harvest; Human; Human immunodeficiency virus test; Image Analysis; Immune; Impaired cognition; In Vitro; Incidence; Individual; Infant; Infection; Institutes; Jordan; Lead; Life; Life Expectancy; Link; Longevity; Medical; Medical Research; Methods; Microglia; Microscopy; Monitor; Nerve Degeneration; Neuraxis; Neurites; Neurocognitive Deficit; Neurodevelopmental Disorder; Neuroglia; Neurologic Effect; Neurologic Symptoms; Neuronal Differentiation; Neurons; Oxidative Stress; Pathway interactions; Patients; Penetrance; Pennsylvania; Performance; Pharmacologic Substance; Phase; Preclinical Testing; Pregnant Women; Process; Protocols documentation; Rattus; Research; Research Contracts; Research Personnel; Rest; Rodent; Safety; Schizophrenia; Science; Standardization; Synapses; System; Tenofovir; Testing; Therapeutic; Thinking; Toxic effect; Universities; antiretroviral therapy; assay development; autism spectrum disorder; biological adaptation to stress; blood-brain barrier crossing; cell type; cerebral atrophy; digital; drug discovery; efavirenz; emtricitabine; epigenetic marker; high throughput screening; in vitro Model; induced pluripotent stem cell; microscopic imaging; nerve stem cell; neurocognitive disorder; neurodevelopment; neurogenesis; neuroinflammation; neuron loss; neurotoxicity; novel; novel therapeutics; pre-clinical; pre-clinical research; preservation; prevent; prophylactic; side effect; stem cells; targeted agent; voltage

Globally, about 40 million people live with HIV/AIDS, and HIV antiretroviral agents (ARVs) are a multi-billion dollar pharmaceutical sector with continued development of agents. Thanks to combination antiretroviral therapy (cART, in which 2 to 4 ARVs are given, simultaneously) HIV+ people have a near-normal life-span. HIV+ pregnant women also receive cART, and their children are given cART prophylactically at birth (in general, cART is given to all HIV+ infants and children). For all HIV+ patients, if cART is stopped, HIV replication restarts; thus, HIV+ people must remain on cART for the rest of their lives. For HIV+ adults, ARVs are present during aging, in which loss of central nervous system (CNS) neurons contributes to loss of cognition/dementia. For HIV+ infants and children, ARVs are present during neurodevelopment. HIV+ individuals frequently develop HIV-Associated Cognitive Disorders (HAND), which includes HIV-associated dementia (HAD,common in AIDS), and less severe forms (asymptomatic neurocognitive impairment and mild neurocognitive disorder (MND). While cART has reduced the incidence of HAD, ANI and MND remain at high levels, even when HIV load is very low. In fact, certain ARVs may contribute to HAND, and alarmingly, HIV+children on cART exhibit developmental delays in intellectual performance. Also, HIV infects/replicates in microglia (MG), an innate immune cell of the CNS, and MG are out of the reach of ARVs that are poor at crossing the blood:brain barrier. More brain-penetrant ARVs were developed, but, unexpectedly, these are linked to greater incidence of HAND. Preclinical research on ARV neurotoxicity has been conducted on primary rodent neurons and glial. Our goal is to develop an assay system (dubbed the hCNS-HIV/ARV platform) for testing ARVs for neurotoxicity and efficacy at inhibiting HIV, neurons, MG and astrocytes (another glial cells), derived from human induced pluripotent stem cells (hiPSC-neurons, hiPSC-MG, and hiPSC-ACs). In phase I, we developed methods for testing ARVs on hiPSC-neurons plated in 384-well dishes, utilizing high-throughput digital microscopy/analysis. We found neurotoxicity (reductions in neurites, synapses, and calcium transients) for elvitegravir (EVG, strongest effect, matching data with rat neurons), dolutegravir (DTG, linked to birth defects), and tenofovir disoproxil fumerate (TDF) and certain combinations of ARVs representing cART had stronger effects than the ARVs, alone. TDF also reduced the viability and elicited epigenetic changes in human neural precursor cells (hNPCs), suggesting that TDF may affect neurogenesis, a process critical for neurodevelopment and cognition. In phase II we will develop standard assay protocols/analysis methods for testing ARVs and HIV infection, itself, on hiPSC-neurons/-MG/-ACs, and on hiPSC-NPCs. The hCNS- HIV/ARV platform will enable development of ARVs that inhibit HIV with minimal neurotoxicity and will be marketed to pharmaceutical companies developing novel therapeutics for HIV/AIDs.

在全球范围内，大约有4000万人患有艾滋病毒/艾滋病，艾滋病毒抗逆转录病毒药物（ARV）是数十亿人 Dollar Pharmaceutical行业，代理商持续开发。多亏了抗逆转录病毒的组合 治疗（手推车，同时给予2至4个ARV）HIV+患者的生命近期跨度。 艾滋病毒+孕妇也接受购物车，他们的孩子在出生时就可以预防手推车（在 将一般的手推车送给所有艾滋病毒+婴儿和儿童）。对于所有艾滋病毒+患者，如果停止购物，艾滋病毒 复制重新启动；因此，艾滋病毒+人必须在余生中留在购物车上。对于艾滋病毒+成年人，ARVS 在衰老期间存在，其中中枢神经系统（CNS）神经元的丧失有助于丧失 认知/痴呆症。对于艾滋病毒+婴儿和儿童，神经发育过程中存在ARV。艾滋病毒+ 个人经常发展与HIV相关的认知障碍（手），其中包括与HIV相关的 痴呆症（在艾滋病中常见），形式不太严重（无症状的神经认知障碍和轻度 神经认知障碍（MND）。虽然购物车降低了HAD的发生率，但ANI和MND仍保持高 水平，即使艾滋病毒负荷非常低。实际上，某些ARV可能有助于手动，令人震惊地 艾滋病毒+儿童在购物车中表现出智力表现的发展延迟。另外，艾滋病毒感染/重复 小胶质细胞（MG），CNS的先天免疫细胞和Mg不超出ARV的范围 越过血液：脑屏障。开发了更多的脑渗透ARV，但出乎意料的是，这些是 与更大的手相关。关于原发性ARV神经毒性的临床前研究 啮齿动物神经元和神经胶质。我们的目标是开发一个分析系统（称为HCNS-HIV/ARV平台） 测试ARV在抑制HIV，神经元，MG和星形胶质细胞（另一个神经胶质细胞）的神经毒性和功效 源自人类诱导的多能干细胞（HIPSC-神经元，HIPSC-MG和HIPSC-ACS）。在第一阶段， 我们开发了使用高通量的384孔菜肴测试ARV的方法 数字显微镜/分析。我们发现神经毒性（神经突，突触和钙瞬态的减少） 对于ElviteGravir（EVG，最强的效果，与大鼠神经元的匹配数据），DoluteGravir（DTG，与出生有关 缺陷）和替诺福韦毒素烟雾（TDF）和代表购物车的某些ARV的组合已 单独的效果比ARV更强。 TDF还降低了人类的生存力和引起的表观遗传变化 神经前体细胞（HNPC），表明TDF可能影响神经发生，这是一个至关重要的过程 神经发育和认知。在第二阶段，我们将开发标准测定方案/分析方法 在HIPSC-神经元/-MG/-ACS以及HIPSC-NPC上测试ARV和HIV感染本身。 HCNS- 艾滋病毒/ARV平台将使以最小的神经毒性抑制艾滋病毒的ARV开发，并将是 销售给开发艾滋病毒/艾滋病新型治疗剂的制药公司。