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 克氏锥虫（T. cruzi 或 Tc）是恰加斯心肌病的病原体，广泛分布于 拉丁美洲和美国南半部估计有超过 800 万例病例，约 1.2 亿例。 拉丁美洲的高危人群。宿主免疫反应对该疾病的结果至关重要。 (Mphis) 作为免疫系统的控制开关，维持亲和抗之间的平衡 在恰加斯病的情况下，外周和组织发挥着重要作用。 然而，Mphis 未能消除寄生虫，并进展至慢性疾病阶段。 Tc 致病分离株劫持宿主先天性和适应性免疫系统的机制，以确保 它在宿主中的存活很大程度上是未知的。对宿主病原体生物学的深入了解。 相互作用对于制定针对 Tc 的预防和治疗对策非常重要 感染。 长非编码 RNA（lncRNA，˃ 200 个核苷酸）包括多种不编码的 RNA 其中，一类与RNA结合蛋白（RBP）结合的lncRNA被认为发挥着关键作用。 lncRNA-RBP 是否在转录后/翻译机制和蛋白质合成的调节中发挥作用。 相互作用塑造宿主免疫仍然是控制寄生虫感染的一个尚未探索的机会。 通过对 lncRNA 阵列的系统分析和验证方法，我们鉴定了以下转录本变体 1-3 lncRNA Morrbid 在感染 Tc 的 Mφ 中高度上调。 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 lncRNA Morrbid-3 与锌指蛋白 36 (ZFP36) RBP 结合。 Tc 感染的 Mφ 增加，RNAi 介导的 Morrbid-3 表达操纵增强 促炎细胞因子/趋化因子信号通路并导致细胞内寄生虫的清除。 本提案要测试的中心假设是 Tc 利用 lncRNA Morrbid- 的调节 我们将使用 3/ZFP36 RBP 复合物来减弱先天免疫并确保其在宿主细胞内存活。 分子生物学和免疫学方法以独立但机械相关的方式检验这一假设 在目标 1 中，我们的目标是确定 lncRNA Morrbid-3 是否具有抗凋亡和免疫调节作用。 在目标 2 中，Morrbid-3 的消耗促进了寄生虫的清除。 Morrbid-3/ZFP36 复合物介导 Mφ 响应 Tc 感染的免疫调节机制。 拟议研究的影响将是发现lncRNA Morrbid-的新机制 3/ZFP36 RBP 复合物决定 Tc 感染的结果，重要的是，我们将确定是否是小分子。 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.