Modeling CNS dynamics in HIV infection and cannabinoids with forebrain organoids

用前脑类器官模拟 HIV 感染和大麻素中的中枢神经系统动力学

• 批准号: 10432007
• 负责人: Kimberly Christian
• 金额: $ 69.52万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10432007
• 关键词: 3-Dimensional; Acute; Adherence; Adult; Animals; Antiinflammatory Effect; Astrocytes; Autopsy; Brain region; Cannabinoids; Cell Communication; Cells; Central Nervous System Diseases; Cerebral cortex; Cerebrum; Chronic; Coculture Techniques; Communicable Diseases; Complement; Data Set; Development; Disease; Disease model; Drug Addiction; Drug abuse; Early Diagnosis; Early Intervention; Electrophysiology (science); Exhibits; Exogenous Factors; Exposure to; Genetic; Genetic Transcription; Glutamates; Goals; HIV; HIV Infections; HIV Seropositivity; HIV-associated neurocognitive disorder; Heterogeneity; Human; Immune system; Immunologics; Impaired cognition; Impairment; Individual; Investigation; Kinetics; Measures; Mediating; Microglia; Modeling; Molecular Profiling; Morphology; Neuraxis; Neuroimmune; Neurons; Organoids; Outcome; Pathogenesis; Pathology; Patients; Pharmaceutical Preparations; Phenotype; Physiological; Play; Population; Predisposition; Property; Prosencephalon; Quality of life; Regimen; Repression; Resources; Role; Severities; Somatic Cell; Specificity; Structure; Symptoms; Synapses; System; Technology; Testing; Therapeutic; Therapeutic Intervention; Time; Tissues; Tropism; Viral; Viral Load result; Viral Proteins; Viral reservoir; Virulence; Virus Replication; Work; antiretroviral therapy; base; behavioral impairment; biological systems; cell type; combat; comorbidity; differentiation protocol; directed differentiation; efficacy evaluation; endocannabinoid signaling; experimental study; human disease; improved; induced pluripotent stem cell; macrophage; marijuana use; motor impairment; neuroinflammation; neuron development; neuropathology; neurotoxic; novel; phytocannabinoid; protein expression; recruit; relating to nervous system; response; self organization; single-cell RNA sequencing; stem cell model; three dimensional cell culture; three dimensional structure; three-dimensional modeling; transcriptome; transcriptome sequencing; translational approach; virus host interaction

PROJECT SUMMARY HIV-Associated Neurocognitive Disorder (HAND) persists in up to 50% of the HIV-positive population, often in spite of low or undetectable viral loads due to effective antiretroviral therapy (ART). Microglia are the resident macrophages in the central nervous system (CNS) and are thought to be a persistent reservoir of HIV in ART treated patients. Microglia secrete a panoply of factors that impact neuronal function and these secreted factors are altered during HIV infection of microglia. In addition to HIV virus-host interactions mediated by microglia, many exogenous factors can modify these interactions or directly impact the CNS itself, making it difficult to dissect the underlying causes of neural pathology leading to HAND. To establish a physiologically- relevant platform to investigate the dynamics of HIV infection in the context of ART and other co-factors, we will use human induced pluripotent stem cells to generate human microglia that we can co-culture with our well-characterized 3D model of the developing cerebral cortex. This integrated 3D cerebral organoid model will recapitulate many of the critical cell populations thought to contribute to HAND-related pathology including glutamatergic neurons, astrocytes, and microglia. We will evaluate the efficacy of the cerebral organoids to model HIV infections by measuring HIV replication kinetics and suppression, as well as HIV protein expression in different cell types (Aim 1). We will then measure the impact of HIV on neuronal development and function at several time points following acute exposure (Aim 2). We will perform morphological and electrophysiological analyses and RNA sequencing at both the single cell and population level to determine the cell type-specific transcriptional responses to HIV exposure. Finally, we will evaluate the impact of exogenous drugs on these measures of viral replication and neuronal function (Aim 3). As a proof-of-concept, we will test a preferred regimen of ART, as well as cannabinoids, both with and without HIV exposure to begin to dissect the impact of these common co-factors on cellular properties that may contribute to pathology underlying HAND. Successful completion of these experiments will result in a validated platform to model the dynamics of neuroinflammatory disorders using brain region-specific cerebral organoids integrated with human microglia.

项目概要 HIV 相关神经认知障碍 (HAND) 持续存在于高达 50% 的 HIV 阳性人群中，通常发生在 尽管由于有效的抗逆转录病毒治疗（ART），病毒载量较低或无法检测到。小胶质细胞是居民 中枢神经系统 (CNS) 中的巨噬细胞被认为是 ART 中 HIV 的持久储存库 接受治疗的患者。小胶质细胞分泌一系列影响神经元功能的因子，这些因子 HIV感染小胶质细胞期间因素发生改变。除了由 HIV 介导的病毒与宿主相互作用外， 小胶质细胞中，许多外源因素可以改变这些相互作用或直接影响中枢神经系统本身，使其 很难剖析导致 HAND 的神经病理学根本原因。建立生理学—— 相关平台来研究 ART 和其他辅助因素背景下的 HIV 感染动态，我们 将使用人类诱导多能干细胞来生成人类小胶质细胞，我们可以将其与我们的共培养 发育中的大脑皮层的特征良好的 3D 模型。这种集成的 3D 大脑类器官模型将 概括了许多被认为有助于 HAND 相关病理学的关键细胞群，包括 谷氨酸能神经元、星形胶质细胞和小胶质细胞。我们将评估大脑类器官的功效 通过测量 HIV 复制动力学和抑制以及 HIV 蛋白表达来建立 HIV 感染模型 在不同的细胞类型中（目标 1）。然后我们将测量艾滋病毒对神经元发育和功能的影响 急性暴露后的几个时间点（目标 2）。我们将进行形态学和电生理学检查 在单细胞和群体水平上进行分析和 RNA 测序，以确定细胞类型特异性 HIV暴露的转录反应。最后，我们将评估外源性药物对这些的影响 病毒复制和神经元功能的测量（目标 3）。作为概念验证，我们将测试首选 ART 疗法以及大麻素，无论是否接触过 HIV，都开始剖析其影响 这些共同的细胞特性辅助因素可能导致 HAND 的病理学。成功的 这些实验的完成将产生一个经过验证的平台来模拟神经炎症的动力学 使用与人类小胶质细胞整合的大脑区域特异性脑类器官来治疗疾病。