Structural and molecular dissection of NF-kappaB regulation by the ubiquitin E3 ligase PDLIM2 in lung innate immunity and diseases

肺先天免疫和疾病中泛素 E3 连接酶 PDLIM2 调节 NF-kappaB 的结构和分子解析

• 批准号: 10586099
• 负责人: Zhaoxia Qu
• 金额: $ 47.85万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-10 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10586099
• 关键词: Acetylation; Acute Lung Injury; Age; Automobile Driving; Bacteria; Binding; Biochemical; Biological Assay; Cell Line; Cells; Cessation of life; Chronic Obstructive Pulmonary Disease; Clinical; Clinical Treatment; Combined Modality Therapy; Complex; Crystallography; Data; Deacetylation; Disease; Dissection; Endotoxins; Epithelial Cells; Epithelium; Exhibits; Functional disorder; Genetic Transcription; Host Defense; Human; Immunology procedure; In Vitro; Incidence; Infection; Inflammation; Inflammatory; Interstitial Lung Diseases; Investigation; Knock-in Mouse; Knock-out; Knockout Mice; LIM Domain; Lipopolysaccharides; Lung; Lung Neoplasms; Lung diseases; Lung infections; Malignant - descriptor; Malignant neoplasm of lung; Mediating; Modification; Molecular; Morbidity - disease rate; Mus; NF-kappa B; Natural Immunity; Nuclear; Outcome; Pathogenesis; Pathogenicity; Pathologic; Patients; Pattern; Physiological; Physiology; Play; Predisposition; Proteins; RELA gene; Regulation; Repression; Ring Finger Domain; Role; Safety; Severities; Severity of illness; Signal Transduction; Stimulus; Testing; Time; Tumor Suppressor Proteins; Ubiquitination; United States; Woman; chromatin immunoprecipitation; conditional knockout; fighting; human model; improved; in vivo; innovation; lung injury; lung tumorigenesis; member; men; mimetics; mortality; mouse model; neoplastic cell; new therapeutic target; novel; novel therapeutics; prevent; recruit; single-cell RNA sequencing; ubiquitin-protein ligase

Abstract NF-B plays a causative role in the inflammation and pathogenesis of various diseases such as lung disease, the third leading killer in the United States responsible for one in seven deaths. However, we have been unable to successfully target it for clinical treatment due to its equally important roles in physiology, and in particular, innate immunity and host defense. Teasing apart these functions of NF-B will overcome this barrier resulting in a powerful means to fight lung and other diseases. Although the core mechanism driving NF-B activation has been well defined and is the same under most physiological and pathogenic conditions, the mechanistic difference in physiologic versus pathogenic NF-B remains largely unknown. Recently, we have demonstrated, for the first time, that NF-B exhibits different activation patterns in normal and malignant lung epithelial cells, the first line of defense and a key component of the innate immunity in the lung. Furthermore, we have revealed, also for the first time, the PDZ-LIM domain-containing protein PDLIM2 as a tumor suppressor and ubiquitin E3 ligase that selectively degrades the ‘pathogenic’ form but not the ‘physiologic’ form of NF-B (thereby preventing pathogenic activation while allowing physiologic activation of NF-B by inflammatory stimuli) and can be targeted as mono- or combination therapy in authentic mouse models of human lung cancer. Like in human lung cancer, of note, PDLIM2 is repressed in the lungs of patients with chronic obstructive pulmonary disease (COPD) or interstitial lung diseases (ILDs), and PDLIM2 repression is associated with disease severity and poor patient survival. Lung epithelial-specific or global deletion of PDLIM2 renders mice highly susceptible to spontaneous and induced lung cancers as well as acute lung injury and death by the bacteria endotoxin lipopolysaccharide (LPS). Based on these trailblazing discoveries, in this proposal we will identify the functional partners of PDLIM2 and determine the structural and biochemical mechanisms by which they act as a ubiquitin E3 ligase complex to dichotomize the differential activation of NF-B in lung epithelial cells. We will also determine in vivo and in vitro the roles and molecular mechanisms of this regulation in lung disease and host defense against pulmonary infection using conditional and inducible knockout (KO) or knock-in (KI) mice and cells of PDLIM2 and/or NF-B. These studies will improve our understanding of normal lung physiology and pulmonary diseases, and open new avenues to study NF-B regulation and action. They may also lead to new clinically feasible approaches to selectively target pathogenic NF-B and reveal new therapeutic targets for better lung disease treatment.

抽象的 NF-B在各种疾病（例如肺部疾病）的炎症和发病机理中起致病作用 美国的第三名领位杀手负责七分之一的死亡。但是，我们一直无法 由于其在生理学中同样重要的作用，特别是 先天免疫和宿主防御。嘲笑NF-B的这些功能将克服这一障碍 以一种与肺部和其他疾病作斗争的强大手段。虽然驱动NF-B激活的核心机制 在大多数物理和致病条件下，机械条件已定义得很好，并且在大多数物理和致病条件下相同 生理与致病性NF-B的差异在很大程度上未知。最近，我们已经证明了 NF-B首次在正常和恶性肺上皮细胞中表现出不同的激活模式， 第一道防线和肺部先天免疫的关键组成部分。此外，我们已经透露， 同样，含PDZ-LIM结构域PDLIM2作为肿瘤抑制剂和泛素E3，这也是第一次 连接酶有选择地降解“致病性”形式，而不是nf-b的“生理”形式（从而防止） 致病激活，同时允许通过炎症刺激对NF-B进行生理激活），并且可以针对目标 作为人类肺癌的真实小鼠模型中的单或联合疗法。像在人类肺癌中一样 值得注意的是，PDLIM2反映在慢性阻塞性肺疾病（COPD）或 间质性肺部疾病（ILD）和PDLIM2表达与疾病严重程度和患者差有关 生存。 PDLIM2的肺上皮特异性或全球删除使小鼠非常容易受到赞助 并诱发肺癌以及急性肺部损伤和死亡。 （LPS）。基于这些开拓性的发现，在此提案中，我们将确定PDLIM2的功能合作伙伴 并确定它们充当泛素E3连接酶复合物的结构和生化机制 二分法在肺上皮细胞中NF-的差异激活。我们还将确定体内和体外 该调节在肺部疾病中的作用和分子机制和宿主防御肺部的防御 使用条件和诱导敲除（KO）或敲除（KI）小鼠以及PDLIM2和/或NF-B的细胞感染。 这些研究将提高我们对正常肺部生理学和肺部疾病的理解，并开放新的 研究NF-B调节和行动的途径。它们也可能导致新的临床可行方法 有选择地靶向致病性NF-B，并揭示了新的治疗靶标，以改善肺部疾病治疗。