Utilizing Single Cell Biological Approaches to Understand CNS TB

利用单细胞生物学方法了解中枢神经系统结核

基本信息

批准号：
9817044
负责人：
Chris G Dulla
金额：
$ 15.35万
依托单位：
TUFTS UNIVERSITY BOSTON
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-25 至 2021-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9817044
关键词：
Address Africa African Astrocytes Autoimmune Diseases Bioinformatics Biological Brain Brain Pathology Cause of Death Cells Central Nervous System Infections Clinical Communicable Diseases Competence Complex Cranial nerve palsies Disease Educational Curriculum Ensure Faculty Functional disorder Future Genetic Transcription Genomic approach Genomics Goals Grant HIV HIV Infections HIV/TB Heterogeneity Human Immune Immune response Immune system Immunocompetence Immunocompetent Immunocompromised Host Immunological Models Immunosuppression Impaired cognition Individual Industrialization Infection Inflammation Inflammatory Inflammatory Response Infrastructure Intervention Investigation Investments Lead Lung diseases Mediating Meningeal Tuberculosis Microglia Modeling Molecular Mus Mycobacterium tuberculosis Neuroglia Neuroimmune Neurologic Dysfunctions Neuronal Dysfunction Neurons Outcome Pathway interactions Patients Process Program Development Regulation Reporting Research Research Infrastructure Resolution Risk Role Scientist Seizures Signal Pathway Signal Transduction South Africa South African Stroke Structure Students TNF gene Therapeutic Intervention Tissues Training Transferable Skills Treatment Factor Tuberculosis Universities Virulent Work body system cell type co-infection design experimental study immunosuppressed improved insight mind control mortality mouse model neuroinflammation neuropathology novel pathogen prevent response single-cell RNA sequencing skill acquisition

项目摘要

PROJECT SUMMARY Mycobacterium tuberculosis (Mtb) is the causative pathogen in tuberculosis (TB). TB is the leading cause of death from infectious disease globally and is especially prevalent in individuals infected with HIV. While normally thought of as a respiratory disease, TB also infects other organ systems. Infection of the central nervous system (CNS-TB) is the most severe form of the disease, and has a mortality rate of nearly 50%, despite aggressive clinical intervention. CNS-TB is associated with severe neurological dysfunction including cranial nerve palsies, cognitive impairment, stroke, and seizures. The brain’s molecular, cellular, and network level response to CNS-TB is almost completely unknown. We hypothesize CNS-TB leads to significant activation of neuroinflammatory signaling, as well as glial and neuronal dysfunction. The risk of developing CNS-TB is markedly increased under conditions of immune suppression, evident in HIV infected individuals, who have a higher occurrence of TB meningitis (TBM). A regulated tumor necrosis factor (TNF) response is a critical feature of immune competence necessary for protection against TB, and is lost in progressive HIV infection. The importance of a proper TNF response is clinically validated by the reactivation of TB in patients on anti-TNF treatment for autoimmune diseases. We recently reported that TNF deficiency (a model of immune suppression) in mice (TNF-/-), promotes CNS-TB infection, a hyper-inflammatory response, gross brain pathology, and mortality. We will utilize a realistic mouse model of CNS-TB in which active Mycobacterium tuberculosis (Mtb) will be injected into the brains of control and immune compromised (TNF-/-) mice to study how resident CNS cell respond. Although Mtb primarily infects microglia and astrocytes, human and mouse neurons also act as host cells for Mtb. Therefore, multiple CNS cells react both directly and indirectly to brain infection, creating a complex cellular- and tissue-level response. To deal with this complexity, we will utilize single cell RNA-seq, a cutting edge genomic approach, which allows transcriptional analysis of the response to CNS-TB on a cell-by-cell basis. This approach enables the identification of individual types of cells, for example astrocytes, and analysis of their unique transcriptional response. In addition, single cell RNA-seq allows investigation of the heterogeneity of the cellular response to CNS-TB by analyzing individual cells. We will utilize single cell RNA-seq to determine how the lack of a proper immune response regulates neuroinflammatory signaling and leads to broad-scale disruption of CNS resident cells in a mouse model of CNS-TB. In doing so, we will establish a partnership between The University of Cape Town and Tufts University. Emphasis will be placed on training South African scientists in neuro- immune interactions, single cell RNA-seq, and advanced genomic analysis approaches, thereby helping develop robust research infrastructure in South Africa.

项目概要结核分枝杆菌 (Mtb) 是结核病 (TB) 的致病病原体。全球范围内死于传染病的人数在艾滋病毒感染者中尤其普遍。结核病通常被认为是一种呼吸道疾病，但它也会感染其他中枢器官系统。神经系统结核（CNS-TB）是该病最严重的形式，死亡率接近 50%，积极的临床干预尽管与严重的神经功能障碍有关，包括脑神经麻痹、认知障碍、中风和癫痫发作。大脑的分子、细胞和网络。对 CNS-TB 的反应水平几乎完全未知，我们追求 CNS-TB 导致显着的结果。神经炎症信号的激活以及神经胶质和神经元功能障碍。在免疫抑制的情况下，中枢神经系统结核的发展风险显着增加，这在艾滋病毒中很明显结核性脑膜炎 (TBM) 受调节的肿瘤坏死因子的发生率较高的感染者。 (TNF) 反应是预防结核病所必需的免疫能力的一个关键特征，并且在进行性 HIV 感染的适当 TNF 反应的重要性通过重新激活得到了临床验证。我们最近报道了 TNF 缺乏症（a）。免疫抑制模型）小鼠（TNF-/-），促进中枢神经系统结核感染，一种高炎症反应，我们将利用一个真实的中枢神经系统结核小鼠模型，其中活跃。结核分枝杆菌 (Mtb) 将被注射到对照和免疫受损 (TNF-/-) 的大脑中尽管结核分枝杆菌主要感染小胶质细胞和星形胶质细胞，但人类却研究了驻留的中枢神经系统细胞如何反应。小鼠神经元也充当结核分枝杆菌的宿主细胞，因此，多个中枢神经系统细胞既直接又发生反应。间接导致大脑感染，产生复杂的细胞和组织水平反应来应对这一问题。复杂性，我们将利用单细胞 RNA-seq，这是一种尖端的基因组方法，它允许转录这种方法可以逐个细胞地分析对 CNS-TB 的反应。个别类型的细胞，例如星形胶质细胞，并分析其独特的转录反应。此外，单细胞 RNA-seq 可以通过以下方式研究细胞对 CNS-TB 反应的异质性：我们将利用单细胞 RNA-seq 来分析单个细胞，以确定如何缺乏适当的免疫。反应调节神经炎症信号传导并导致中枢神经系统驻留细胞的大规模破坏在此过程中，我们将与开普大学建立合作伙伴关系。镇和塔夫茨大学将重点培训南非科学家。免疫相互作用、单细胞 RNA-seq 和先进的基因组分析方法，从而帮助南非发展强大的研究基础设施。