A new brain organoid model for NeuroHIV and the impact of opioids

NeuroHIV 的新脑类器官模型以及阿片类药物的影响

• 批准号: 10693976
• 负责人: Alysson R. Muotri
• 金额: $ 78.83万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10693976
• 关键词: Adherent Culture; Animal Model; Astrocytes; Biopsy; Blood; Brain; Brain Diseases; Buprenorphine; Cardiac Myocytes; Cell Line; Cells; Central Nervous System; Complex; Deacetylation; Disease; Enzymes; Epidemic; Event; Exposure to; Functional disorder; Gene Expression; Generations; Genomics; Goals; HIV; HIV Infections; HIV-1; Histone Acetylation; Histone Deacetylation; Human; Immune; Immune response; Immunologic Markers; In Vitro; Individual; Infection; Integration Host Factors; Learning; Link; Mediating; Memory; Methods; Microglia; Modeling; Modification; Molecular; Morphine; Neurologic; Neuronal Dysfunction; Neuronal Plasticity; Neurons; Neuropathogenesis; Opioid; Opioid Rotation; Organoids; Outcome; Pathogenesis; Pathway interactions; Patients; Persons; Pharmaceutical Preparations; Phenotype; Predisposition; Protocols documentation; Risk; Site; Stem Cell Development; System; Technology; Time; Tissue-Specific Gene Expression; Tissues; Viral; Viremia; Virus Replication; advanced disease; antiretroviral therapy; brain dysfunction; brain tissue; buprenorphine treatment; cell type; experimental study; extracellular; human model; human pluripotent stem cell; induced pluripotent stem cell; innovation; innovative technologies; methadone treatment; morphometry; neural network; neuroAIDS; neuroinflammation; neuropathology; opioid exposure; opioid use; self assembly; single-cell RNA sequencing; success; synaptogenesis; three dimensional structure; transcriptome

Abstract HIV-1 infects resident cells of the central nervous system (CNS) leading to neuropathogenesis. HIV- neuropathogenesis is likely caused by direct and indirect viral and host factors. However, the exact underlying mechanisms remain unclear. Despite the success of antiretroviral therapy (ART) in suppressing HIV replication, near half of people living with HIV (PLWH) still have varying degrees of HIV-associated neurological disorders (HAND). There is also evidence that the CNS serves as an HIV reservoir and sanctuary site that may allow low level viremia, contributing to persistent neuroinflammation. The evolving molecular events underlying HIV neuropathogenesis are difficult to delineate, partially due to the lack of realistic HIV animal models and because human brain tissues rarely become available for studies until patients die, often due to advanced diseases. Human brain cortical organoids (BCO) are an emerging, cutting-edge technology for studying neuropathological disorders; because of their human origin, they better match the genomic and structural features of the developing human brain compared to animal models. This model consists of a self-assembled dynamic 3-D structure that provides an interplay of different cell types, which is limited in traditional monolayer cultures. We optimized protocols to generate long-term viable and functional BCO. Our BCO model has an unprecedented cell type diversity, via a dynamic development from progenitor cells to neuronal cells, that become interspersed with quiescent astrocytes over time; a difficult phenotype to obtain ex vivo. With the cellular components for generation of a functional neural network in place, our BCO model shows a robust extracellular electrical activity at early stages and progressively develops into an organized oscillatory network. Additionally, we have previously established methods for integration of iPSC-derived microglia into the BCO forming an assembloid, which is crucial tor this study. A BCO assembloid model containing relevant immune cell types will enable susceptibility to HIV and the study of the contributions of different cell types to the neurological consequences of infection. Using this robust and functional BCO assembloid, we propose to develop a new human model to study the cellular and molecular mechanisms underlying HIV neuropathogenesis, and the potential interactive, additive, or synergistic effects of antiretroviral treatment (ART) and opioid exposure. This microglia-infused BCO with endogenous astrocytes will allow HIV infection and its related pathophysiological events and help to disentangle the contribution and interplay of relevant immune cells to neuropathogenesis.

抽象的 HIV-1感染中枢神经系统（CNS）的居民细胞，导致神经病发生。艾滋病病毒- 神经病发生可能是由直接和间接病毒和宿主因素引起的。但是，确切的基础 机制仍然不清楚。尽管抗逆转录病毒疗法（ART）在抑制HIV复制方面取得了成功，但 近一半的艾滋病毒（PLWH）仍然患有不同程度的艾滋病毒相关神经系统疾病 （手）。也有证据表明，中枢神经系统用作艾滋病毒水库和庇护所，可能允许较低 水平的病毒血症，导致持续性神经炎症。 HIV的基础分子事件不断发展 神经病发生很难描述，部分原因是缺乏现实的HIV动物模型，并且因为 人类脑组织很少用于研究，直到患者死亡，通常是由于晚期疾病。 人脑皮质器官（BCO）是一种研究神经病理学的新兴，尖端的技术 疾病；由于其人类的起源，它们更好地匹配了发展中的基因组和结构特征 与动物模型相比，人脑。该模型由一个自组装的动态3-D结构组成 提供了不同细胞类型的相互作用，该类型在传统的单层文化中受到限制。我们优化了 生成长期可行和功能性BCO的协议。我们的BCO模型具有前所未有的细胞类型 通过从祖细胞到神经元细胞的动态发展，多样性变得散布 随着时间的流逝，星形胶质细胞静止；一个难以获得离体的表型。带有细胞成分 我们的BCO模型生成功能性神经网络，显示出强大的细胞外电活动 在早期阶段，并逐步发展为有组织的振荡网络。另外，我们还有 以前已建立的方法将IPSC衍生的小胶质细胞整合到BCO中，形成一个汇编， 这是这项研究至关重要的。包含相关免疫细胞类型的BCO组装模型将启用 对艾滋病毒的敏感性以及对不同细胞类型对神经系统后果的贡献的研究 感染。使用这种强大且功能性的BCO组装，我们建议开发一种新的人类模型来研究 HIV神经病发生的基础的细胞和分子机制，以及潜在的相互作用， 抗逆转录病毒治疗（ART）和阿片类药物暴露的添加剂或协同作用。这个注入小胶质细胞的BCO 使用内源性星形胶质细胞将允许HIV感染及其相关的病理生理事件，并有助于 解散相关免疫细胞对神经病的作用的贡献和相互作用。