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.

项目摘要 与HIV相关的神经认知障碍（Hand）持续多达50％的HIV阳性人群，通常在 由于有效的抗逆转录病毒疗法（ART）而导致低或无法检测的病毒负荷。小胶质细胞是居民 中枢神经系统（CNS）中的巨噬细胞，被认为是艺术中艾滋病毒的持续水库 治疗的患者。小胶质细胞分泌一个影响神经元功能的因素，这些因素分泌 小胶质细胞感染期间因素改变了因素。除了由HIV病毒宿主相互作用介导 小胶质细胞，许多外源性因素可以改变这些相互作用或直接影响CNS本身，使其成为 难以剖析神经病理的根本原因导致手。在生理上建立 在艺术和其他共同因素的背景下研究艾滋病毒感染动态的相关平台，我们 将使用人类诱导的多能干细胞来产生人类的小胶质细胞，我们可以与我们的培养 发育中的大脑皮层的特征良好的3D模型。该集成的3D大脑器官模型将 概括了许多关键细胞群体被认为有助于与手相关的病理 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，模型HIV感染模型 在不同的细胞类型中（AIM 1）。然后，我们将衡量HIV对神经元发育和功能的影响 急性暴露后几个时间点（AIM 2）。我们将执行形态学和电生理学 分析和RNA测序在单细胞和种群水平上都确定细胞类型特异性 对HIV暴露的转录反应。最后，我们将评估外源性药物对这些药物的影响 病毒复制和神经元功能的度量（AIM 3）。作为概念验证，我们将测试一个首选 艺术方案以及大麻素，无论是在有和没有艾滋病毒的情况下，都开始剖析 这些在细胞特性上的共同系数可能有助于病理学的基础手。成功的 这些实验的完成将导致一个经过验证的平台来建模神经炎症的动力学 使用与人类小胶质细胞整合的大脑区域特异性大脑器官的疾病。