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（与HIV-1相关） 即使在抑制性抗逆转录病毒疗法（ART）的患者中，神经认知障碍或手）也是如此。作为人们 患有HIV-1年龄的生活，正在发生复合效果，并增加了与年龄相关的痴呆症 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。手的发展方式以及如何发展 经过修改仍然是神秘的，主要是因为研究HIV-1感染并在 人脑。在过去的几年中，干细胞技术的新发展允许 “大脑器官”与诱导多能干细胞（IPSC）的分化，这些培养物可以是 在促进神经外胚层的三维扩张的条件下，体外生长 或“微型大脑”形式。大脑器官是异质的，形成了各种大脑区域，包括 腹前脑，大脑皮层，海马以及中间和后脑边界。他们表现出神经元 具有功能性并且能够进行电兴奋，并发展小胶质细胞。这些脑器官也类似于 基因表达水平的人类皮质发育，并允许对神经网络，细胞进行深入分析 行为，药物筛查，疾病建模和大脑发育的变化。而大脑区域 来自不同IPSC系的类器官中的组成品种，区域基因表达模式在很大程度上保持不变 在个人之间可再现。这些创造了无与伦比的新机会来研究HIV-1的感染 脑。我们建议开发我们的IPSC衍生的类器官模型，该模型纳入了小胶质细胞，该模型是 更好地代表一个可以支持强大的HIV-1感染的成年成熟大脑。使用此模型，我们可以测试 不同的病毒是否导致差异性神经毒性以及小胶质细胞以外的其他细胞是否可以潜在 感染病毒。我们将测试病毒中的单个蛋白质（包括HIV-1 TAT）如何引起神经系统 损害以及诸如Didehydro-Cortistin A（DCA）等艺术或药物，跨性别的TAT活性， 影响过程。有趣的是，暴露于TAT还潜在可卡因介导的奖励机制， 这进一步促进了手，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害方面进行合作，并确定是否是否 DCA可以扭转它。由于脑器官提供了大脑中HIV-1潜伏期的模型，我们可以测试是否是否 DCA的给药可以“阻塞”任何残留病毒，最后是否HIV-1特异性细胞毒性T 细胞（CTL）可以消除器官中的病毒。这些研究一起有望提供新颖的见解 手的发病机理，TAT介导的神经毒性，可卡因的作用以及它们之间的潜在联系 过程。最后，该模型有望在大脑中的HIV-1潜伏期上添加令人兴奋的发现，如果可以的话 沉默或淘汰。

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

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.

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

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