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

项目摘要 结核病（TB），由细胞内细菌病原体分枝杆菌引起 结核病（M.tb）仍然是世界上最致命的传染病之一。 根据世界卫生组织的年度报告，三分之一的人口受到感染， 结核分枝杆菌在世界各地蔓延，导致2019年有1000万例活动性结核病例和140万例死亡。 大约5-10%的感染者在其一生中发展为活动性结核病。的 结核分枝杆菌-宿主相互作用的机制仍有待确定。宿主长非编码RNA lncRNA是具有大于200个核苷酸的非编码RNA。我们的初步数据 和以前的出版物证明了宿主lncRNA参与宿主防御 对抗巨噬细胞中的分枝杆菌感染。然而，研究有限， 阐明宿主lncRNA在结核分枝杆菌感染中的功能。长期 本研究的目的是鉴定对宿主防御有益的宿主lncRNA 抗结核杆菌感染的巨噬细胞，并进一步了解机制， lncRNA作为顺式或反式调节元件介导宿主免疫， 巨噬细胞响应结核分枝杆菌感染。在目前的提案中，我们假设， lncRNA是结核分枝杆菌感染者体内抗分枝杆菌应答的重要组成部分 巨噬细胞为了验证这一假设，我们提出了一个全基因组筛选宿主 人类中调节宿主防御分枝杆菌感染的lncRNA 使用CRISPR-Cas9转录激活系统靶向巨噬细胞， 一万个宿主lncRNA目的1筛选提高宿主存活率的基因间lncRNAs THP-1细胞在结核分枝杆菌感染过程中。目标2将验证候选宿主lncRNA， 在结核分枝杆菌感染的过程中基因工程化的THP-1细胞。核酸- 基础治疗正在成为治疗未满足的 医疗条件。我们的研究将为核酸作为一种新的发展提供线索 用于治疗TB，特别是耐药TB的宿主导向疗法。

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.