Mechanisms of IncRNA/RBP regulation of macrophage function for control of T. cruzi infection

IncRNA/RBP调节巨噬细胞功能控制克氏锥虫感染的机制

• 批准号: 10392964
• 负责人: Imran Hussain Chowdhury
• 金额: $ 19.75万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-15 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10392964
• 关键词: 3' Untranslated Regions; Acute; Adaptive Immune System; Agreement; Anti-Inflammatory Agents; Apoptosis; Apoptotic; Area; Attention; Attenuated; Binding; Binding Proteins; Biological; Biological Assay; Biology; Cell physiology; Cells; Cessation of life; Chagas Disease; Chronic Disease; Chronic Phase of Disease; Clinical; Code; Complex; Data Set; Development; Disease; Disease Outcome; Elements; Enhancers; Ensure; Equilibrium; Eukaryota; Exhibits; Frequencies; Gene Expression; Genes; Genetic Transcription; Genome; Genomic DNA; Immune; Immune response; Immune system; Immunity; Immunologic Surveillance; Immunology; Immunoprecipitation; Infection; Inflammation; Inflammatory; Inflammatory Response; Invaded; Knowledge; Latin America; Longevity; Macrophage Activation; Mediating; Messenger RNA; Molecular; Molecular Biology; Molecular Immunology; Mus; Muscle Cells; Myeloid Cells; Natural Immunity; Nucleotides; Organ; Outcome; Oxidative Stress; Parasites; Parasitic infection; Pathogenicity; Peripheral; Persons; Play; Post-Transcriptional Regulation; Preventive treatment; Protein Biosynthesis; Proteins; RNA; RNA Binding; RNA Interference; RNA Splicing; RNA-Binding Proteins; Regulation; Reporting; Research; Ribonucleoproteins; Risk; Role; Shapes; Signal Pathway; Signal Transduction; Testing; Therapeutic; Tissues; Transcript; Trypanosoma cruzi; Untranslated RNA; Variant; Zinc Fingers; adaptive immunity; chagasic cardiomyopathy; chemokine; crosslink; cytokine; dark matter; genomic RNA; helicase; immune clearance; immunoregulation; improved; innovation; insight; knock-down; mRNA Decay; mRNA Precursor; mRNA Stability; macrophage; molecular sequence database; monocyte; mouse model; mutant; novel; novel therapeutics; overexpression; pathogen; prevent; progenitor; programs; protein complex; response; screening; small molecule inhibitor

ABSTRACT Trypanosoma cruzi (T. cruzi or Tc), the causative agent of Chagas cardiomyopathy, is widely distributed in Latin America and the southern half of the USA. It is estimated that >8 million recorded cases and ~ 120 million people at risk in Latin America. The host immune response is critical for outcome of the disease. Macrophages (Mϕs) function as control switches of the immune system and maintain balance between pro- and anti- inflammatory response. In context to Chagas disease, peripheral and tissue Mϕs play an essential role against acute Tc infection. However, Mϕs fail to eliminate the parasite, and progress to chronic disease phase. The mechanisms by which pathogenic isolates of Tc hijack the host innate and adaptive immune system to ensure its survival in the host are largely unknown. An in-depth understanding of the biology of host-pathogen interactions is important for the development of preventative and treatment countermeasures against Tc infection. Long non-coding RNAs (lncRNAs, ˃ 200 nucleotides) include a diverse class of RNAs that do not encode proteins. Among them, a class of lncRNA that binds to RNA binding protein (RBP) is regarded to play a critical role in regulation of post-transcriptional / translational machinery and protein synthesis. Whether lncRNA-RBP interactions shape host immunity remains an underexplored opportunity for control of parasitic infection. Through systematic analysis of lncRNA arrays and a confirmatory approach, we have identified transcript variants 1-3 of lncRNA Morrbid as highly upregulated in Mφs infected with Tc. Unbiased screening of cross-linking immunoprecipitation-sequencing databases and subsequent RNA immunoprecipitation (RIP) assays demonstrated that lncRNA Morrbid-3 binds to Zinc finger protein 36 (ZFP36) RBP. ZFP36 expression was increased in Mφs infected by Tc, and RNAi mediated manipulation of Morrbid-3 expression strengthened proinflammatory cytokine/chemokine signaling pathways and resulted in clearance of intracellular parasites. The central hypothesis to be tested in this proposal is that Tc utilizes the regulation of lncRNA Morrbid- 3/ZFP36 RBP complex to attenuate innate immunity and ensure its intracellular survival in host. We will employ molecular biology and immunology approaches to test this hypothesis in independent, yet mechanistically related aims. In Aim 1, our objectives are to determine if lncRNA Morrbid-3 exerts anti-apoptotic and immunomodulatory effects, and depletion of Morrbid-3 promotes parasite clearance by Mφ. In Aim 2, we will examine the molecular mechanism of Morrbid-3/ZFP36 complex mediated immune regulation of Mφ in response to Tc infection. The impact of the proposed studies will be discovery of novel mechanisms by which lncRNA Morrbid- 3/ZFP36 RBP complex determines the outcomes of Tc infection. Importantly, we will determine if small molecule inhibitors of Morrbid-3/ZFP36 interaction will offer novel therapeutic avenues to control Tc infection.

抽象的 Crypanosoma Cruzi（T. Cruzi或TC），Chagas心肌病的致病药物，广泛分布在 拉丁美洲和美国南部。据估计，> 800万案件和约1.2亿 拉丁美洲有风险的人。宿主免疫反应对于疾病的结局至关重要。巨噬细胞 （M ϕ）作为免疫系统的控制开关的功能，并保持亲和抗 炎症反应。在chagas病的背景下，外围和组织m ϕs扮演着重要的角色 急性TC感染。但是，M ϕ无法消除寄生虫，并发展为慢性病阶段。这 TC的致病性分离株劫持宿主先天和适应性免疫系统的机制，以确保 它在宿主中的生存在很大程度上是未知的。对宿主病原体生物学的深入了解 相互作用对于开发针对TC的预防和治疗对策很重要 感染。 长的非编码RNA（LNCRNA，˃200个核苷酸）包括不编码的RNA的潜水员类别 蛋白质。其中，与RNA结合蛋白（RBP）结合的一类LNCRNA被认为起关键 在调节转录后 /翻译机制和蛋白质合成中的作用。 LNCRNA-RBP是否 相互作用形状的宿主免疫仍然是控制寄生感染的毫无创伤的机会。通过 LNCRNA阵列的系统分析和确认方法，我们已经确定了成绩单变体1-3 lncRNA morrbid在感染TC的Mφ中高度更新。交联的无偏筛选 免疫沉淀 - 测序数据库和随后的RNA免疫沉淀（RIP）测定法 证明lncRNA morrbid-3与锌指蛋白36（ZFP36）RBP结合。 ZFP36表达是 在被TC感染的Mφ中增加，RNAi介导的Morrbid-3表达的操纵增强了 促炎性细胞因子/趋化因子信号通路，并导致细胞内寄生虫的清除。 该提议中要检验的中心假设是TC利用了lncrna morrbid-的调节 3/ZFP36 RBP复合物可减弱先天免疫并确保其在宿主中的细胞内存活。我们将雇用 分子生物学和免疫学方法在独立但机械相关的独立但机械相关的方法中检验这一假设 目标。在AIM 1中，我们的目标是确定LNCRNA Morrbid-3是否导出抗凋亡和免疫调节 Morrbid-3的效果和耗尽可促进Mφ的寄生虫清除。在AIM 2中，我们将检查分子 Morrbid-3/ZFP36复合物介导的Mφ的免疫调节的机理，响应TC感染。 拟议的研究的影响将是发现lncrna morrbid-的新机制 3/ZFP36 RBP复合物确定TC感染的结果。重要的是，我们将确定是否小分子 Morrbid-3/ZFP36相互作用的抑制剂将为控制TC感染提供新的治疗途径。

项目成果

Oxidative stress implications for therapeutic vaccine development against Chagas disease.

• DOI: 10.1080/14760584.2021.1969230
• 发表时间: 2021-11
• 期刊: Expert review of vaccines
• 影响因子: 6.2
• 作者: Choudhuri S;Rios L;Vázquez-Chagoyán JC;Garg NJ
• 通讯作者: Garg NJ

Acute muscle mass loss was alleviated with HMGB1 neutralizing antibody treatment in severe burned rats.

• DOI: 10.1038/s41598-023-37476-4
• 发表时间: 2023-06-24
• 期刊: SCIENTIFIC REPORTS
• 影响因子: 4.6
• 作者: Song, Juquan;Chowdhury, Imran H.;Choudhuri, Subhadip;Ayadi, Amina E. I.;Rios, Lizette E.;Wolf, Steven E.;Wenke, Joseph C.;Garg, Nisha J.
• 通讯作者: Garg, Nisha J.