Development of Brain Organoids to Study the Impact of HIV-1, Drugs of Abuse and Aging on Cognitive Impairment

开发大脑类器官来研究 HIV-1、滥用药物和衰老对认知障碍的影响

• 批准号: 10613440
• 负责人: DOUGLAS F NIXON
• 金额: $ 73.75万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-15 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10613440
• 关键词: 3-Dimensional; Acceleration; Address; Adult; Affect; Age; Aging; Alkaloids; Autologous; Bioinformatics; Brain; Brain region; CCL2 gene; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; Cell Line; Cell physiology; Cells; Cellular Immunology; Cerebral cortex; Cerebrum; Cicatrix; Cocaine; Collaborations; Complex; Cytotoxic T-Lymphocytes; Dementia; Development; Disease model; Drug Addiction; Drug Screening; Effectiveness; Effector Cell; Engineering; Exhibits; Exposure to; FOXO3A gene; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Goals; HIV; HIV-1; HIV-associated neurocognitive disorder; Hand; Healthcare Systems; Heterogeneity; Hippocampus; Human; Image; Immune; Impaired cognition; In Vitro; Individual; Infection; Interdisciplinary Study; Interleukin-1 beta; Internet; Lead; Link; Mediating; Methods; Microglia; Microscopy; Modeling; Molecular Virology; Nervous System Trauma; Neurocognitive Deficit; Neuroectoderm; Neuroglia; Neurons; Organoids; Pathogenesis; Pathway interactions; Peripheral Blood Mononuclear Cell; Persons; Pharmaceutical Preparations; Phenotype; Process; Prosencephalon; Reporter; Reproducibility; Residual state; Rewards; Role; Signaling Protein; Structure; Substance abuse problem; System; TNF gene; Technology; Testing; Toxic effect; Variant; Viral Proteins; Virus; aging brain; analog; antiretroviral therapy; cell behavior; cell killing; cognitive testing; combinatorial; cortistatin; drug of abuse; hindbrain; induced pluripotent stem cell; innovation; insight; longevity gene; mouse model; neural network; neuroimmunology; neuroinflammation; neurotoxicity; nonhuman primate; novel; novel strategies; novel therapeutics; single cell analysis; stem cell technology; stimulant abuse; tat Protein; transcriptome; transcriptome sequencing; uptake

PROJECT ABSTRACT HIV-1 associated neuroinflammation and neurotoxicity lead to cognitive impairments (HIV-1-associated neurocognitive disorders or HAND) even in those under suppressive antiretroviral therapy (ART). As people living with HIV-1 age, a compounding effect is occurring, with age associated dementia added to HAND, leading to a complex web of neurocognitive deficit. This will have tremendous implications for health care systems not only in the USA but also in the developing world. How HAND develops, and how it could be modified remain mysterious, largely because It has been very difficult to study HIV-1 infection and HAND in the human brain. Over the past few years new developments in stem cell technologies have permitted the differentiation of “cerebral organoids” from induced pluripotent stem cells (iPSCs), and these cultures can be grown in vitro in conditions that promote three-dimensional expansion of neuroectoderm, in cerebral organoid or “miniature brain” forms. Cerebral organoids are heterogeneous and form a variety of brain regions, including ventral forebrain, cerebral cortex, hippocampus, and mid- and hindbrain boundary. They exhibit neurons that are functional and capable of electrical excitation, and develop microglia. These brain organoids also resemble human cortical development at the gene expression level, and allow in depth analysis of neural networks, cell behavior, drug screening, disease modeling, and variations in brain development. While brain-region composition varies in organoids from different iPSC lines, regional gene-expression patterns remain largely reproducible across individuals. These create unparalleled new opportunities to study HIV-1 infection of the brain. We propose to develop our iPSC derived organoid model which incorporates microglia, into one that better represents an adult mature brain that can support robust HIV-1 infection. With this model, we can test whether different viruses lead to differential neurotoxicity and if cells other than microglia can be latently infected with virus. We will test how individual proteins from the virus, including HIV-1 Tat causes neurological damage, and how ART or drugs such as Didehydro-Cortistatin A (dCA), which cross-neutralizes Tat activity, affects the process. Interestingly, exposure to Tat also potentiates cocaine-mediated reward mechanisms, which further promotes HAND, revealing a complex web of interactions between HIV-1 infection and drugs of substance abuse. We will determine how Tat and cocaine collaborate in neurological damage and determine if dCA can reverse it. As cerebral organoids provide a model for HIV-1 latency in the brain we can test whether administration of dCA, can “block-and-lock” any residual virus, and finally whether HIV-1 specific cytotoxic T cells (CTL) can eliminate virus in the organoid. Together, these studies promise to provide novel insights into the pathogenesis of HAND, Tat mediated neurotoxicity, effects of cocaine, and the potential link between these processes. Finally, the model promises to add exciting findings on HIV-1 latency in the brain, and if it can be silenced or eliminated.

项目摘要 HIV-1相关的神经炎症和神经毒性导致认知障碍（HIV-1相关 即使在接受抑制性抗逆转录病毒治疗（ART）的患者中，也会出现神经认知障碍（HAND）。作为人 与 HIV-1 年龄一起生活，复合效应正在发生，与年龄相关的痴呆症被添加到 HAND 中， 导致复杂的神经认知缺陷网络。这将对医疗保健产生巨大影响 系统不仅在美国，而且在发展中国家。 HAND 是如何发展的，以及它会如何发展 修改仍然是个谜，很大程度上是因为在人类中研究 HIV-1 感染和 HAND 非常困难 人类的大脑。在过去的几年里，干细胞技术的新发展使得 从诱导多能干细胞（iPSC）分化出“大脑类器官”，这些培养物可以 在促进大脑类器官中神经外胚层三维扩张的条件下体外生长 或“微型大脑”形式。大脑类器官具有异质性，形成多种大脑区域，包括 腹侧前脑、大脑皮层、海马以及中脑和后脑边界。他们表现出的神经元 有功能并能够电激发，并发育小胶质细胞。这些大脑类器官也类似于 人类皮质发育在基因表达水平上，并允许深入分析神经网络、细胞 行为、药物筛选、疾病建模和大脑发育的变化。而大脑区域 来自不同 iPSC 系的类器官的组成有所不同，区域基因表达模式在很大程度上仍然存在 可在个体之间复制。这些为研究 HIV-1 感染创造了无与伦比的新机会 脑。我们建议开发我们的 iPSC 衍生的类器官模型，该模型将小胶质细胞纳入其中 更好地代表了可以支持强大的 HIV-1 感染的成人成熟大脑。通过这个模型，我们可以测试 不同的病毒是否会导致不同的神经毒性，以及小胶质细胞以外的细胞是否可以潜伏 感染病毒。我们将测试病毒中的单个蛋白（包括 HIV-1 Tat）如何引起神经系统疾病 损害，以及 ART 或诸如二脱氢皮质抑素 A (dCA) 之类的药物如何交叉中和 Tat 活性， 影响进程。有趣的是，接触 Tat 也会增强可卡因介导的奖励机制， 这进一步促进了 HAND，揭示了 HIV-1 感染与药物之间复杂的相互作用网络 药物滥用。我们将确定 Tat 和可卡因如何在神经损伤中协同作用，并确定是否 dCA 可以逆转它。由于大脑类器官提供了大脑中 HIV-1 潜伏期的模型，我们可以测试是否 给予 dCA，可以“阻断并锁定”任何残留病毒，最后确定 HIV-1 特异性细胞毒性 T 细胞（CTL）可以消除类器官中的病毒。总之，这些研究有望提供新的见解 HAND 的发病机制、Tat 介导的神经毒性、可卡因的作用以及这些因素之间的潜在联系 流程。最后，该模型有望增加有关大脑中 HIV-1 潜伏期的令人兴奋的发现，以及是否可以 被压制或消除。

项目成果

The chaperone protein p32 stabilizes HIV-1 Tat and strengthens the p-TEFb/RNAPII/TAR complex promoting HIV transcription elongation.

• DOI: 10.1073/pnas.2217476120
• 发表时间: 2023-01-03
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Li, Chuan;Mori, Luisa P.;Lyu, Shuang;Bronson, Ronald;Getzler, Adam J.;Pipkin, Matthew E.;Valente, Susana T.
• 通讯作者: Valente, Susana T.

Next-Generation Human Cerebral Organoids as Powerful Tools To Advance NeuroHIV Research.

• DOI: 10.1128/mbio.00680-21
• 发表时间: 2021-08-31
• 期刊: mBio
• 影响因子: 6.4
• 作者: Premeaux TA;Mediouni S;Leda A;Furler RL;Valente ST;Fine HA;Nixon DF;Ndhlovu LC
• 通讯作者: Ndhlovu LC

HIV-1 Remission: Accelerating the Path to Permanent HIV-1 Silencing.

• DOI: 10.3390/v15112171
• 发表时间: 2023-10-28
• 期刊: Viruses
• 作者: Lyons DE;Kumar P;Roan NR;Defechereux PA;Feschotte C;Lange UC;Murthy N;Sameshima P;Verdin E;Ake JA;Parsons MS;Nath A;Gianella S;Smith DM;Kallas EG;Villa TJ;Strange R;Mwesigwa B;Furler O'Brien RL;Nixon DF;Ndhlovu LC;Valente ST;Ott M
• 通讯作者: Ott M

Mapping disease regulatory circuits at cell-type resolution from single-cell multiomics data.

• DOI: 10.1038/s43588-023-00476-5
• 发表时间: 2023-07
• 期刊: NATURE COMPUTATIONAL SCIENCE
• 作者: Chen, Xi;Wang, Yuan;Cappuccio, Antonio;Cheng, Wan-Sze;Zamojski, Frederique Ruf;Nair, Venugopalan D.;Miller, Clare M.;Rubenstein, Aliza B.;Nudelman, German;Tadych, Alicja;Theesfeld, Chandra L.;Vornholt, Alexandria;George, Mary-Catherine;Ruffin, Felicia;Dagher, Michael;Chawla, Daniel G.;Soares-Schanoski, Alessandra;Spurbeck, Rachel R.;Ndhlovu, Lishomwa C.;Sebra, Robert;Kleinstein, Steven H.;Letizia, Andrew G.;Ramos, Irene;Fowler Jr, Vance G.;Woods, Christopher W.;Zaslavsky, Elena;Troyanskaya, Olga G.;Sealfon, Stuart C.
• 通讯作者: Sealfon, Stuart C.

Transcriptional regulation of the HIV-1 inhibitory factor human mannose receptor 1 by the myeloid-specific transcription factor PU.1.

骨髓特异性转录因子 PU.1 对 HIV-1 抑制因子人甘露糖受体 1 的转录调节。

• DOI: 10.1128/jvi.01702-23
• 发表时间: 2024
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Mallorson,Rosa;Miyagi,Eri;Kao,Sandra;Sukegawa,Sayaka;Saito,Hideki;Fabryova,Helena;MorellattoRuggieri,Luciana;Mediouni,Sonia;Valente,SusanaT;Strebel,Klaus
• 通讯作者: Strebel,Klaus