Targeting non-coding RNAs as host-directed drug therapy for tuberculosis

项目总结 结核分枝杆菌是结核病(TB)的病原体，每年导致160万人死亡。 交配感染结核分枝杆菌。有一个迫切的需要 开发新的治疗方法。在这个项目中，我们旨在研究宿主非编码的作用 核糖核酸(NcRNA)在非洲两个研究地点(开普敦)对结核病易感性的驱动作用 南非小镇和乌干达坎帕拉)。聚焦于人类原代巨噬细胞，我们计划整合， 利用系统生物学的方法，研究具有编码和非编码基因表达的遗传结构 对结核分枝杆菌感染的反应情况。我们将确定反应表达数量性状基因座(ReQTL) 用于编码基因、长非编码RNA(LncRNAs)和将与之整合的microRNAs(miRNAs 编码/非编码基因表达、共表达和放松管制网络，以确定关键变体和 调节巨噬细胞对结核分枝杆菌感染的反应。最有希望的miRNAs和lncRNAs 将通过正交技术进行验证，并考虑在人类巨噬细胞中实现功能化。 在这里，我们将使用反义寡核苷酸在体外感染结核分枝杆菌的功能丧失方法。 评估人单核细胞来源的巨噬细胞对细胞内分枝杆菌生长和 机械巨噬细胞特异性抗分枝杆菌效应功能的研究。瞄准这些人 已识别的宿主miRNAs和lncRNAs有望成为辅助宿主导向药物的候选分子 结核病的治疗。 我们的中心假设是：a)Mtb劫持宿主miRNAs和lncRNAs以建立成功的 巨噬细胞的持久性和存活率，b)人类变异可以影响关键角色及其作用，以及 C)可以利用对这一点的理解来识别、设计、实施和测试基于RNA的新型 抗菌治疗策略。这一假设建立在我们之前未发布和已发布的数据基础上 [1]显示Mtb靶向巨噬细胞中的miR-143、miR-365和lincRNA-MIR99AHG促进 分枝杆菌通过微调宿主基因的表达水平来生长，并得到越来越多的支持 其他研究的结果[2-16]。我们的长期目标是开发一种针对宿主的反义治疗药物系统， 特别是抑制在结核病发病机制中至关重要的miRNAs和lncRNAs。其基本原理是 这一方法将创造一种新的、有针对性的、以宿主为导向的、基于RNA的结核病药物疗法。要实现 为了实现这一目标，我们组建了一支由ncRNA生物学、传染病、系统生物学和 来自南部非洲、乌干达和美国多个地点的生物信息学。

PROJECT SUMMARY Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), causes 1.6 million deaths per mated to be infected with Mtb. There is an urgent need to develop new treatment approaches. In this project, we aim to investigate the role of the host non-coding RNA (ncRNA) landscape in driving susceptibility to tuberculosis (TB) across two study sites in Africa (Cape Town, South Africa and Kampala, Uganda). Focusing on human primary macrophages, we plan to integrate, using systems biology approaches, genetic architecture with the coding and non-coding gene expression landscape of response to Mtb infection. We will identify response expression quantitative trait loci (ReQTLs) for coding genes, long non-coding RNAs (lncRNAs), and microRNAs (miRNAs) which will be integrated with coding/non-coding gene expression, co-expression, and deregulation networks to prioritize key variants and regulators orchestrating macrophage response to Mtb infection. The most promising miRNAs and lncRNAs will be validated with orthogonal techniques and considered for functionalization in human macrophages. Here, we will employ loss-of function approaches using antisense oligonucleotides in ex vivo Mtb-infected human monocyte-derived macrophages to evaluate the outcome on intracellular mycobacterial growth and investigation of mechanistic macrophage-specific anti-mycobacterial effector functions. Targeting these identified host miRNAs and lncRNAs could become promising candidates for adjunctive host-directed drug therapy for TB. Our central hypothesis is that a) Mtb hijacks host miRNAs and lncRNAs to establish successful persistence and survival in macrophages, b) that human variation can affect key players and their roles, and c) that an understanding of this can be leveraged to identify, design, implement, and test a novel RNA-based anti-microbial treatment strategy. This hypothesis builds upon our previous unpublished and published data [1] showing that Mtb targets miR-143, miR-365 and lincRNA-MIR99AHG in macrophages to promote mycobacterial growth by fine-tuning expression levels of host genes and is supported by a growing number of other studies [2-16]. Our long-term goal is to develop a host-directed drug system for antisense therapy, specifically inhibiting miRNAs and lncRNAs that are crucially involved in TB pathogenesis. The rationale for this approach would be to create a novel, targeted host-directed RNA-based drug therapy for TB. To achieve this goal, we have assembled a team of experts in ncRNA biology, infectious disease, systems biology, and bioinformatics from multiple sites in Southern Africa, Uganda and the USA.