Examining HIV-mediated disruption of CNS immune homeostasis using a triple humanized mouse

摘要 目前，缺乏足够的动物模型来复制人类HIV-1感染的事件， 是研究中枢神经系统（CNS）HIV病理学的关键障碍。根除艾滋病毒1型感染 已成为当务之急。当代的高效抗逆转录病毒疗法（ART）显著延长了 艾滋病毒感染者的生命，但它没有解决艾滋病毒相关的神经认知障碍（HAND）的发病率。 HIV持续存在于人类血液淋巴隔室和中枢神经系统（CNS）中。持久性 外周和中枢神经系统中的艾滋病毒宿主会导致慢性炎症和终末器官疾病。临床 观察结果显示病毒存在于CNS驻留免疫细胞和CNS的非免疫细胞中， 如星形胶质细胞。CNS病毒储库是终身的。尽管如此，关于持续性CNS病毒感染的性质的知识 水库和他们的影响，神经免疫稳态是有限的，因为缺乏现有的小 动物模型我们的目的是建立一种新的TRIPLE人源化小鼠模型， 人血液/血淋巴系统，小胶质细胞和星形胶质细胞，研究外周HIV- 人脑胶质细胞在发育中CNS免疫和神经元的感染和免疫激活 功能障碍同时具有外周血/免疫细胞和人脑细胞的小鼠（HuBB-小鼠）是 有望为研究神经艾滋病提供一个全面的模型。该模型最适合于研究 属于当前RFA范围内的研究领域。使用新的HuBB小鼠，我们的目标是研究：1）。艾滋病毒 抑制性ART期间的胶质细胞储库和使用单细胞的神经免疫稳态失调 转录组学分析; 2）.神经元的完整性和功能的改变，作为失调的结果， 神经免疫稳态用ART治疗的动物在所有组织和细胞中抑制外周病毒复制 类型，包括星形胶质细胞和小胶质细胞，促进病毒学，免疫学和神经病理学评估。 这项拟议的工作预计将为新的HuBB小鼠奠定基础，用于未来评估多种 慢性感染/免疫激活背景下的HAND病理学方面， CNS和神经炎症。小鼠模型将成为开发治疗方法以减轻 HAND和设计从CNS储库中根除HIV的策略。

ABSTRACT Currently, the lack of adequate animal models to replicate the events of HIV-1 infection in humans presents a critical barrier to study HIV pathology of the central nervous system (CNS). Eradicating HIV-1 infection has become a priority. Contemporary, highly active antiretroviral therapy (ART) has significantly prolonged the lives of those infected, but it has not resolved the incidence of HIV-associated neurocognitive disorders (HAND). HIV persists in the human hemato-lymphoid compartments and in the central nervous system (CNS). Persistent HIV reservoirs both in the periphery and CNS result in chronic inflammation and end-organ diseases. Clinical observations reveal that the virus is present in CNS-resident immune cells and in non-immune cells of the CNS, such as astrocytes. The CNS viral reservoir is life-long. Still, knowledge on the nature of persistent CNS viral reservoirs and their influence on neuroimmune homeostasis is limited because of deficiencies in existing small animal models. We aim to establish a new TRIPLE humanized mouse model reconstituted with autologous human blood/ hematolymphoid system, microglia, and astrocytes to study the interaction of peripheral HIV- infection and immune activation with human brain glial cells in developing CNS immune and neuronal dysfunction. Mice that possess both peripheral blood/immune cells and human brain cells (HuBB-mice) are expected to provide a comprehensive model to study NeuroAIDS. The model is best suitable to study the research areas that fall within the scope of the current RFA. Using the new HuBB-mice, we aim to study: 1). HIV glial reservoirs during suppressive ART and the dysregulation of neuroimmune homeostasis using single-cell transcriptomic analysis, and 2). the altered neuronal integrity and function as a consequence of dysregulated neuroimmune homeostasis. Animals treated with ART suppress peripheral viral replication in all tissues and cell types, including astrocytes and microglia, facilitating virologic, immunologic, and neuropathologic assessments. The proposed work is expected to lay the groundwork on the new HuBB-mice for future evaluations of multiple aspects of HAND pathology in the context of chronic infection/ immune activation, persistent HIV reservoirs in CNS, and neuroinflammation. The mouse model will be an important tool to develop therapeutics to alleviate HAND and for designing HIV eradication strategies from CNS reservoirs.