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

项目摘要 结核分枝杆菌（Mtb）是结核病（TB）的病原体，每年造成160万人死亡。 交配后感染结核分枝杆菌。迫切需要 开发新的治疗方法。在这个项目中，我们的目标是调查的作用，主机非编码 在非洲（开普敦）的两个研究地点，RNA（ncRNA）在驱动结核病（TB）易感性方面的作用 镇，南非和坎帕拉，乌干达）。专注于人类初级巨噬细胞，我们计划整合， 利用系统生物学方法，遗传结构与编码和非编码基因表达 对结核病感染的反应情况。我们将鉴定反应表达数量性状基因座（ReQTL） 对于编码基因，长非编码RNA（lncRNA）和微小RNA（miRNAs），它们将与 编码/非编码基因表达，共表达和去调控网络，以优先考虑关键变体， 调控巨噬细胞对Mtb感染的应答。最有前途的miRNAs和lncRNAs 将采用正交技术进行验证，并考虑在人巨噬细胞中进行功能化。 在这里，我们将采用功能丧失的方法，使用反义寡核苷酸在离体结核分枝杆菌感染的 人单核细胞衍生的巨噬细胞，以评价细胞内分枝杆菌生长的结果， 机制巨噬细胞特异性抗分枝杆菌效应子功能的研究。靶向这些 鉴定的宿主miRNAs和lncRNAs可能成为有希望的宿主导向药物的候选者， 治疗结核病。 我们的中心假设是a）Mtb劫持宿主miRNAs和lncRNAs以建立成功的 B）人类变异可以影响关键参与者及其作用，以及 c）对这一点的理解可以用来识别，设计，实施和测试一种新的基于RNA的 抗菌处理策略。这一假设建立在我们以前未发表和已发表的数据基础上 [1]显示Mtb靶向巨噬细胞中的miR-143、miR-365和lincRNA-MIR 99 AHG，以促进巨噬细胞的增殖。 通过微调宿主基因的表达水平来控制分枝杆菌的生长， 其他研究[2-16]。我们的长期目标是开发一种用于反义治疗的宿主导向药物系统， 特异性抑制在TB发病机制中至关重要的miRNA和lncRNA。的理由 这种方法将创造一种新的、靶向宿主的、基于RNA的结核病药物疗法。实现 为了实现这一目标，我们组建了一个由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.