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

全基因组筛选调节宿主防御巨噬细胞分枝杆菌感染的宿主lncRNA

• 批准号: 10526531
• 负责人: Yong Cheng
• 金额: $ 22.44万
• 依托单位: OKLAHOMA STATE UNIVERSITY STILLWATER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-02 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10526531
• 关键词: Address; Annual Reports; Antimycobacterial Agents; Biology; CRISPR-mediated transcriptional activation; CRISPR/Cas technology; Cell Survival; Cell physiology; Cells; Cessation of life; Communicable Diseases; Data; Development; Disease; Drug resistance in tuberculosis; Engineering; Genetic Engineering; Genetic Transcription; Goals; Grant; Host Defense; Human; Human Engineering; Immune response; Immunity; Infection; Knowledge; Libraries; Light; Link; Mammalian Cell; Mediating; Medical; Mycobacterium Infections; Mycobacterium tuberculosis; Nucleic Acids; Nucleotides; Organism; Outcome; Pathway interactions; Persons; Play; Population; Public Health; Publications; Regulatory Element; Reporting; Role; System; Techniques; Testing; Therapeutic; Transcript; Transcriptional Activation; Tuberculosis; Untranslated RNA; base; gain of function; genome wide screen; genome-wide; improved; loss of function; macrophage; microbial; mycobacterial; novel; pathogen; pathogenic bacteria; response; screening; tuberculosis treatment

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.

项目概要 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。 根据世界卫生组织年度报告，三分之一的人口已被感染 结核病在全球范围内蔓延，导致 2019 年有 1000 万活动性结核病例和 140 万人死亡。 大约 5-10% 的感染者在其一生中会患上活动性结核病。这 M.tb-宿主相互作用的机制仍有待定义。宿主长非编码RNA (lncRNA) 是长度超过 200 个核苷酸的非编码 RNA。我们的初步数据 和之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。但研究有限 解决 M.tb 感染背景下宿主 lncRNA 的功能。长期来看 这项研究的目标是鉴定有利于宿主防御的宿主lncRNA 对抗巨噬细胞M.tb感染，进一步了解宿主机制 lncRNA作为顺式或反式调节元件介导宿主免疫 巨噬细胞对结核分枝杆菌感染的反应。在当前的提案中，我们假设 lncRNA 是结核分枝杆菌感染者抗分枝杆菌反应的重要组成部分 巨噬细胞。为了检验这一假设，我们提出了对宿主进行全基因组筛选 调节宿主防御人类分枝杆菌感染的lncRNA 巨噬细胞使用 CRISPR-Cas9 转录激活系统，靶向超过 10,000 个宿主 lncRNA。目标 1 将筛选提高存活率的宿主基因间 lncRNA M.tb 感染期间 THP-1 细胞的变化。目标 2 将验证候选宿主 lncRNA 在 M.tb 感染过程中对 THP-1 细胞进行基因改造。核酸- 基于治疗正在成为治疗未满足治疗的新兴一类新疗法 医疗条件。我们的研究将揭示核酸作为一种新型药物的发展 用于治疗结核病，特别是耐药结核病的宿主导向疗法。