Genome-wide screening for host lncRNAs regulating host defense against Mycobacterial infection in macrophages

项目摘要 结核(TB)，由细胞内细菌病原体分支杆菌引起 结核病(M.TB)仍然是世界上最致命的传染病之一。 根据世卫组织的年度报告，三分之一的人口感染了 结核分枝杆菌在世界各地传播，导致2019年1000万活动性结核病病例和140万人死亡。 大约5%-10%的感染者在他们的一生中患上活动性结核病。这个 结核分枝杆菌与宿主相互作用的机制尚不明确。宿主长的非编码RNA (LncRNAs)是大于200个核苷酸的非编码RNA。我们的初步数据 以前的出版物证明了宿主lncRNAs参与宿主防御 抗巨噬细胞中的分枝杆菌感染。然而，这方面的研究有限。 在结核分枝杆菌感染的背景下阐述宿主LncRNAs的功能。长期的 这项拟议的研究的目标是识别有利于宿主防御的宿主lncRNAs 抗巨噬细胞感染结核分枝杆菌，并进一步了解宿主 LncRNAs作为顺式或跨式调节元件，在介导宿主免疫中发挥作用 巨噬细胞对结核分枝杆菌感染的反应。在目前的提案中，我们假设 LncRNAs是结核分枝杆菌感染体内抗分枝杆菌反应的重要组成部分 巨噬细胞。为了验证这一假设，我们建议对宿主进行全基因组筛查。 调节宿主抵抗分枝杆菌感染的IncRNAs 巨噬细胞使用CRISPR-Cas9转录激活系统靶向 10,000个主机lncRNA。目标1将筛选宿主基因间lncRNAs以提高存活率 在结核分枝杆菌感染过程中THP-1细胞的数量。AIM 2将验证候选宿主LncRNA 结核分枝杆菌感染过程中的基因工程THP-1细胞。核酸- 基础疗法正在成为治疗未成年患者的一种新兴疗法。 医疗条件。我们的研究将有助于揭示核酸作为一种新型材料的发展。 宿主导向疗法治疗结核病，特别是耐药结核病。

Project Summary Tuberculosis (TB), caused by the intracellular bacterial pathogen Mycobacterium tuberculosis (M.tb), remains one of the most deadly infectious diseases in the world. According to the WHO annual report, one third of the population has been infected by M.tb across the world, leading to 10 million active TB cases and 1.4 million deaths in 2019. About 5-10% of infected people develop active TB disease during their lifetime. The mechanism of M.tb-host interactions remains to be defined. Host long noncoding RNAs (lncRNAs) are noncoding RNAs with greater than 200 nucleotides. Our preliminary data and previous publications demonstrated the engagement of host lncRNAs in host defense against mycobacterial infections in macrophages. However, there are limited studies addressing the functions of host lncRNAs in the context of M.tb infection. The long-term goal of this proposed study is to identify host lncRNAs that are beneficial to host defense against M.tb infection in macrophages, and further understand the mechanism how host lncRNAs serve as cis- or trans-regulatory elements in mediating the host immunity in macrophages in response to M.tb infection. In the current proposal, we hypothesize that lncRNAs are essential components of antimycobacterial response within M.tb-infected macrophages. To test the hypothesis, we propose a genome-wide screening for host lncRNAs that regulate host defense against mycobacterial infection in human macrophages using a CRISPR-Cas9 transcription activation system that targets over 10,000 host lncRNAs. Aim 1 will screen host intergenic lncRNAs that improve the survival of THP-1 cells during M.tb infection. Aim 2 will validate candidate host lncRNAs in genetically engineered THP-1 cells during the course of M.tb infection. Nucleic acid- based therapy is becoming an emerging new class of therapeutics for treating unmet medical conditions. Our study will shed light on the development of nucleic acid as a novel host-directed therapy for the treatment of TB, especially drug-resistant TB.