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: 10397158
• 负责人: PATRICK M MCDONOUGH
• 金额: $ 94.1万
• 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 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10397158
• 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; Persons; 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，约有4000万人感染艾滋病毒/艾滋病，艾滋病毒抗逆转录病毒药物（ARV）是数十亿美元， 元医药行业随着代理商的不断发展。多亏了联合抗逆转录病毒疗法 艾滋病病毒阳性者的寿命接近正常。 艾滋病毒阳性孕妇也接受cART，其子女在出生时接受cART治疗（在 一般而言，cART适用于所有HIV阳性婴儿和儿童）。对于所有HIV+患者，如果停止cART， 复制重新开始;因此，HIV+患者必须在其余生中继续接受cART治疗。对于艾滋病毒阳性成人， 存在于衰老过程中，其中中枢神经系统（CNS）神经元的损失导致神经元的损失。 认知/痴呆。对于艾滋病毒阳性的婴儿和儿童，抗逆转录病毒药物存在于神经发育过程中。HIV+ 个人经常发展HIV相关的认知障碍（HAND），其中包括HIV相关的 痴呆症（HAD，常见于艾滋病）和不太严重的形式（无症状神经认知障碍和轻度 神经认知障碍（MND）。虽然cART降低了HAD的发病率，但ANI和MND仍然很高 即使艾滋病毒载量很低。事实上，某些抗逆转录病毒药物可能会导致HAND，令人担忧的是， 接受cART治疗的HIV阳性儿童表现出智力发育迟缓。此外，艾滋病毒感染/复制 小胶质细胞（MG）是中枢神经系统的一种先天性免疫细胞，而MG是ARV无法达到的， 穿过血脑屏障更多的大脑穿透抗逆转录病毒药物被开发出来，但是，出乎意料的是，这些药物 与手的发病率更高有关。ARV神经毒性的临床前研究已在原代小鼠上进行 啮齿动物神经元和神经胶质。我们的目标是开发一个检测系统（称为hCNS-HIV/ARV平台）， 测试ARV的神经毒性和抑制HIV、神经元、MG和星形胶质细胞（另一种神经胶质细胞）的功效， 来源于人诱导多能干细胞（hiPSC-神经元、hiPSC-MG和hiPSC-AC）。在第一阶段， 我们开发了在384孔培养皿中接种的hiPSC神经元上测试抗逆转录病毒药物的方法， 数字显微镜/分析。我们发现神经毒性（神经突、突触和钙瞬变减少） 对于elvitegravir（EVG，最强效应，与大鼠神经元匹配的数据），dolutegravir（DTG，与出生有关 缺陷）和富马酸替诺福韦二异山梨酯（TDF）以及代表cART的某些ARV组合， 比单独使用抗逆转录病毒药物的效果更强TDF还降低了人类的生存力，并引起了表观遗传学的变化。 神经前体细胞（hNPC），表明TDF可能影响神经发生，这是一个关键的过程， 神经发育和认知。在第二阶段，我们将制定标准的测定方案/分析方法， 在hiPSC-神经元/-MG/-AC和hiPSC-NPC上测试ARV和HIV感染本身。hCNS- 艾滋病毒/抗逆转录病毒药物平台将使抗逆转录病毒药物的开发能够以最小的神经毒性抑制艾滋病毒， 销售给开发艾滋病毒/艾滋病新疗法的制药公司。

项目成果

High-content analysis and Kinetic Image Cytometry identify toxicity and epigenetic effects of HIV antiretrovirals on human iPSC-neurons and primary neural precursor cells.

• DOI: 10.1016/j.vascn.2022.107157
• 发表时间: 2022-03
• 期刊: Journal of pharmacological and toxicological methods
• 影响因子: 1.9
• 作者: Smith AS;Ankam S;Farhy C;Fiengo L;Basa RCB;Gordon KL;Martin CT;Terskikh AV;Jordan-Sciutto KL;Price JH;McDonough PM
• 通讯作者: McDonough PM