Mitochondrial DNA and TLR9 in Pulmonary Fibrosis

肺纤维化中的线粒体 DNA 和 TLR9

• 批准号: 10457967
• 负责人: Changwan Ryu
• 金额: $ 16.79万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10457967
• 关键词: Activities of Daily Living; Adult; Area; Biological Assay; Biomedical Engineering; Biometry; Bleomycin; Bronchoalveolar Lavage; Cells; Cellular Assay; Chronic lung disease; Clinical; Collagen; Computational Biology; Department chair; Deposition; Development; Disease; Dose; Environment; Experimental Models; Exposure to; Extracellular Matrix; Fellowship; Fibroblasts; Fibrosis; Funding; Genes; Genomics; Goals; Grant; Harvest; Human; Immune system; Immunologic Receptors; In Vitro; Inflammation; Inflammatory; Inhalation; Interstitial Lung Diseases; Laboratories; Lead; Longitudinal cohort; Lung; Lung Transplantation; Measurement; Mediating; Mentors; Mentorship; Methods; Mitochondria; Mitochondrial DNA; Modeling; Molecular; Mus; Myofibroblast; Natural Immunity; Nature; Outcome; Pathogenesis; Patients; Pattern; Pharmaceutical Preparations; Phenocopy; Phenotype; Physicians; Plasma; Play; Process; Pulmonary Fibrosis; Radiation; Reporter; Research; Research Personnel; Research Project Grants; Role; Scientist; Secondary to; Smooth Muscle Actin Staining Method; Structure of parenchyma of lung; TGFB1 gene; TLR9 gene; Testing; Time; Toxic Environmental Substances; Training; Transforming Growth Factor-Beta Overexpression; Work; alveolar epithelium; antifibrotic treatment; base; career; career development; cell injury; cohort; combat; cost; curative treatments; drug development; epithelial injury; extracellular; fibrotic lung; fibrotic lung disease; idiopathic pulmonary fibrosis; immune activation; in vitro Model; in vivo; inhibitor; insight; mechanotransduction; metabolic profile; mortality; novel; novel therapeutics; overexpression; polyacrylamide hydrogels; response; response to injury; side effect; single-cell RNA sequencing; translational medicine; translational study

PROJECT SUMMARY Candidate: My long-term career goal is to develop a successful academic career and become an independently funded physician-scientist focused on the study of innate immunity in idiopathic pulmonary fibrosis (IPF). Under the guidance of my mentor and research advisors, I have received training in laboratory-based assays, in vitro models of IPF, translational medicine, and biostatistics. I have proposed career development activities that will allow me to continue this training, but also develop unique expertise that is distinct from my mentor in the areas of (1) innate immunity and (2) computational biology. Mentors and Environment: I will be mentored by Dr. Erica Herzog, a globally renowned IPF investigator who has studied the immunopathogensis of fibrosis with an extremely impressive track record of successful mentees. My advisors include Dr. Naftali Kaminski, a well-known IPF investigator whose visionary methods in the genomic profiling of fibrotic lung disease have revolutionized the field; Dr. Wajahat Mehal, who has been lauded for his work with mitochondrial DNA (mtDNA) and Toll-Like Receptor 9 (TLR9) in the development of fibrosis; Dr. Min-Jong Kang, an expert in mitochondrial innate immunity in chronic lung disease; and Dr. Anjelica Gonzalez, who has developed novel methods in bioengineering for ex vivo modeling of the fibrotic microenvironment. My department Chair (Dr. Gary Desir) has assured me that at least 75% of my time will be dedicated to my career development, and he and my Section Chief (Dr. Kaminski) have detailed their commitment. Mentored Research Project: The pathogenesis of IPF involves the uncontrolled accumulation of activated myofibroblasts, which arise in response to TGFβ1 mediated interactions with the stiff fibrotic lung microenvironment. The innate immune receptor TLR9, which recognizes and responds to mtDNA derived from injured cells, has been shown to have a significant role in mediating this process. We showed that mtDNA is released by TGFβ1-stimulated and stiffness-induced normal human lung fibroblasts, where it induces myofibroblast transformation in a manner that phenocopies fibroblasts harvested from the IPF lung. In the clinical setting, mtDNA concentrations are elevated in the plasma of IPF subjects, where it displays a robust association with all-cause mortality in two independent cohorts. Our subsequent studies reveal that mice deficient in TLR9 are protected from fibrosis caused by lung specific overexpression of the bioactive form of the human TGFβ1 gene and by repetitive administration of low-dose inhaled bleomycin. While exciting, however, these studies are limited by not having determined whether mtDNA-induced fibroblast activation requires TLR9, whether TLR9’s fibrosis promoting effects are mediated through fibroblasts in vivo, and the nature of the mtDNA-TLR9 relationship in IPF. Because elucidation of these questions might substantially impact our understanding of pulmonary fibrosis, this K08 application proposes a state-of-the-art set of translational studies to test the hypothesis that mtDNA-TLR9 interaction drives fibroblast activation and fibrosis in the adult lung.

项目摘要 候选人：我的长期职业目标是发展一个成功的学术生涯， 基金资助的医生-科学家专注于特发性肺纤维化（IPF）的先天免疫研究。下 在我的导师和研究顾问的指导下，我接受了基于实验室的体外试验培训， IPF模型、转化医学和生物统计学。我建议开展职业发展活动， 允许我继续这种培训，但也发展独特的专业知识，这是不同于我的导师在这些领域， （1）先天免疫和（2）计算生物学。导师和环境：我将由埃里卡博士指导 Herzog是一位全球知名的IPF研究者，他研究了纤维化的免疫发病机制， 成功学员的卓越记录。我的顾问包括纳夫塔利·卡明斯基博士 IPF研究者，他在纤维化肺病基因组分析方面的富有远见的方法已经彻底改变了 Wajahat Mehal博士因其在线粒体DNA（mtDNA）和Toll样蛋白方面的工作而受到称赞。 受体9（TLR 9）在纤维化发展中的作用; Min-Jong Kang博士，线粒体先天免疫专家 Anjelica Gonzalez博士，他开发了生物工程的新方法， 纤维化微环境的体内建模。我的系主任（加里德西尔博士）向我保证， 我至少75%的时间将用于我的职业发展，他和我的科长（卡明斯基博士） 详细说明了他们的承诺。指导研究项目：IPF的发病机制涉及 活化的肌成纤维细胞不受控制的积聚，这是对TGFβ1介导的相互作用的反应 僵硬的纤维化肺微环境。先天免疫受体TLR 9，它识别并响应 从受损细胞中提取的线粒体DNA，已被证明在介导这一过程中发挥重要作用。我们 结果表明，mtDNA是由TGFβ1刺激和僵硬诱导的正常人肺成纤维细胞释放的， 其中它以表型模仿从IPF收获的成纤维细胞的方式诱导肌成纤维细胞转化 肺。在临床环境中，IPF受试者血浆中mtDNA浓度升高， 在两个独立的队列中，与全因死亡率存在强相关性。我们随后的研究表明， 保护TLR 9缺陷的小鼠免于由生物活性形式的肺特异性过表达引起的纤维化 人TGFβ1基因和重复给予低剂量吸入博莱霉素。虽然令人兴奋， 然而，这些研究受到限制，因为没有确定mtDNA诱导的成纤维细胞活化是否 需要TLR 9，TLR 9的纤维化促进作用是否通过体内成纤维细胞介导，以及 mtDNA-TLR 9与IPF的关系。因为澄清这些问题可能会对我们的 了解肺纤维化，这个K 08申请提出了一个国家的最先进的一套转化研究 以检验mtDNA-TLR 9相互作用驱动成人肺中成纤维细胞活化和纤维化的假设。