Mitochondrial DNA and TLR9 in Pulmonary Fibrosis

项目总结 候选人：我的长期职业目标是发展成功的学术事业，成为一名独立的 受资助的内科医生-科学家专注于特发性肺纤维化(IPF)的先天免疫研究。在……下面 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分析培训 IPF、转化医学和生物统计学的模型。我提出了职业发展活动，这些活动将 请允许我继续进行培训，但也要发展与我的导师在这些领域截然不同的独特专业知识 (1)先天免疫和(2)计算生物学。导师和环境：我将得到埃里卡博士的指导 Herzog是一位全球知名的IPF研究员，他曾与 非常令人印象深刻的成功学员的记录。我的顾问包括Naftali Kaminski博士，著名的 IPF研究人员，他在纤维化肺疾病基因组图谱方面的富有远见的方法带来了革命性的变化 该领域；Wajahat Mehal博士，他因在线粒体DNA(MtDNA)和Toll-Like方面的工作而受到赞扬 受体9(TLR9)在纤维化发展中的作用；线粒体天然免疫专家康敏中博士 以及Anjelica Gonzalez博士，他在生物工程方面为EX开发了新的方法 纤维化微环境的活体模拟。我的系主任(加里·德西尔博士)向我保证 我至少75%的时间将致力于我的职业发展，他和我的科长(卡明斯基博士) 已经详细说明了他们的承诺。指导研究项目：IPF的发病机制涉及 转化生长因子β1介导的相互作用引起的激活的肌成纤维细胞的不受控制的聚集 与僵硬的纤维化肺微环境有关。先天免疫受体TLR9，识别并响应 对来自受损细胞的线粒体DNA，已被证明在这一过程中起着重要的调节作用。我们 研究表明，转化生长因子β1刺激和僵直诱导的正常人肺成纤维细胞释放线粒体DNA， 它诱导肌成纤维细胞转化的方式与从IPF中获取的成纤维细胞的表型相同 阿龙。在临床环境中，ipf受试者血浆中mtdna浓度升高，表现为 在两个独立的队列中，与全因死亡率有很强的相关性。我们随后的研究表明， TLR9基因缺陷的小鼠可免受肺组织中生物活性形式特异性过表达引起的纤维化的影响 人转化生长因子β1基因的表达和重复使用低剂量的博莱霉素。虽然令人兴奋， 然而，这些研究由于没有确定线粒体dna是否诱导成纤维细胞激活而受到限制。 需要TLR9，TLR9的促纤维化作用是否在体内是通过成纤维细胞介导的，以及性质 线粒体DNA-TLR9在IPF中的关系。因为这些问题的澄清可能会对我们的 了解肺纤维化，这个K08应用程序提出了一套最先进的翻译研究 验证线粒体DNA-TLR9相互作用导致成人肺成纤维细胞活化和纤维化的假说。

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.