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: 10529106
• 负责人: Alysson R. Muotri
• 金额: $ 79万
• 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/10529106
• 关键词: Adherent Culture; Animal Model; Astrocytes; Biopsy; Blood; Brain; Brain Diseases; Buprenorphine; Cardiac Myocytes; Cell Line; Cells; Complex; Deacetylation; Disease; Enzymes; Epidemic; Epigenetic Process; 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; Neuraxis; 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 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; 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) 的常驻细胞，导致神经病变。艾滋病病毒- 神经发病机制可能是由直接和间接的病毒和宿主因素引起的。然而，确切的底层 机制仍不清楚。尽管抗逆转录病毒疗法（ART）在抑制艾滋病毒复制方面取得了成功， 近一半的艾滋病毒感染者 (PLWH) 仍患有不同程度的艾滋病毒相关神经系统疾病 （手）。还有证据表明，中枢神经系统是艾滋病毒储存库和避难所，可能会导致低度感染。 病毒血症，导致持续性神经炎症。 HIV 背后不断演变的分子事件 神经发病机制很难描述，部分原因是缺乏现实的艾滋病毒动物模型，而且因为 人类脑组织很少可用于研究，直到患者死亡，通常是由于晚期疾病。 人脑皮质类器官（BCO）是一种用于研究神经病理学的新兴尖端技术 疾病；由于其人类起源，它们更好地匹配发育中人类的基因组和结构特征 人类大脑与动物模型的比较。该模型由一个自组装的动态 3D 结构组成， 提供不同细胞类型的相互作用，这在传统的单层培养中是有限的。我们优化了 生成长期可行且功能性 BCO 的协议。我们的 BCO 模型具有前所未有的细胞类型 多样性，通过从祖细胞到神经元细胞的动态发展，散布着 随着时间的推移，星形胶质细胞处于静止状态；很难在体外获得的表型。与细胞成分 生成功能性神经网络后，我们的 BCO 模型显示出强大的细胞外电活动 在早期阶段逐渐发展成为一个有组织的振荡网络。此外，我们还有 先前建立的将 iPSC 衍生的小胶质细胞整合到 BCO 中形成组合体的方​​法， 这对于这项研究至关重要。包含相关免疫细胞类型的 BCO 组装体模型将能够 对艾滋病毒的易感性以及不同细胞类型对艾滋病毒神经系统后果的贡献的研究 感染。使用这种强大且功能齐全的 BCO 组装体，我们建议开发一种新的人体模型来研究 HIV神经发病机制的细胞和分子机制，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累加或协同作用。这种注入小胶质细胞的 BCO 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 阐明相关免疫细胞对神经发病机制的贡献和相互作用。