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.

摘要 克氏锥虫是查加斯心肌病的病原体，广泛分布于 拉丁美洲和美国的南半部。据估计，记录在案的案例有800万例，约1.2亿例 拉丁美洲的高危人群。宿主的免疫反应对疾病的转归至关重要。巨噬细胞 (MϕS)作为免疫系统的控制开关，维持正反平衡 炎症反应。在恰加斯病的背景下，外周和组织MϕS在防治该病中起着至关重要的作用 急性TC感染。然而，MϕS未能消灭寄生虫，并进展到慢性病阶段。这个 致病菌株劫持宿主固有免疫系统和获得性免疫系统以确保 它在宿主体内的存活在很大程度上是未知的。对寄主病原菌生物学的深入了解 相互作用对制定TC的预防和治疗对策很重要 感染。 长的非编码RNA(LncRNAs，˃200核苷酸)包括不同类别的不编码的RNA 蛋白质。其中，一类与rna结合蛋白(Rbp)结合的lncrna被认为起关键作用。 在转录后/翻译机制和蛋白质合成的调节中的作用。无论lncRNA-RBP 相互作用塑造宿主免疫仍然是控制寄生虫感染的一个未被开发的机会。穿过 对lncRNA阵列的系统分析和确证方法，我们已经确定了转录本变体1-3 在感染了TC的M-φS体内，lncRNA-Morrid高度上调。交联剂的无偏筛选 免疫沉淀-测序数据库和随后的RNA免疫沉淀(RIP)分析 证实了lncRNA Morrid-3与锌指蛋白36(ZFP36)RBP结合。ZFP36的表达是 TC感染M-φS细胞表达增加，RNA干扰对MorrBid-3表达的调控作用增强 促炎症细胞因子/趋化因子信号通路，并导致清除细胞内寄生虫。 在这项提议中要检验的中心假设是，TC利用lncRNA Morrbid- 3/ZFP36 RBP复合体，以减弱天然免疫，确保其在宿主体内的细胞内存活。我们将聘用 分子生物学和免疫学方法在独立但机械相关的情况下检验这一假说 目标。在目标1中，我们的目标是确定lncRNA Morrid-3是否发挥抗细胞凋亡和免疫调节作用。 作用，耗尽MorrBid-3可促进Mφ清除寄生虫。在目标2中，我们将研究分子 MorrBid-3/ZFP-36复合体介导的M-φ对TC感染的免疫调节机制 拟议研究的影响将是发现新的机制，通过这种机制，lncRNA Morrid- 3/ZFP36RBP复合体决定着TC感染的转归。重要的是，我们将确定小分子 Morrid-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.