The role of the epithelial-derived chemokine, CXCL12, in Idiopathic Pulmonary Fibrosis

上皮源性趋化因子 CXCL12 在特发性肺纤维化中的作用

• 批准号: 10383135
• 负责人: Daniel Sullivan
• 金额: $ 6.92万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-16 至 2022-05-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10383135
• 关键词: Acute; Adopted; Adult; Affect; Age; Aging; Animal Model; Animals; Autocrine Communication; Biological Models; Bleomycin; Blood; CXCL12 gene; CXCR4 Receptors; Cell Aging; Cell Line; Cell model; Cells; Cellular biology; Characteristics; Chromosomes; Cicatrix; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; DNA; Data; Development; Diagnosis; Disease; Dominant-Negative Mutation; Elements; Environment; Epithelial; Epithelial Cell Proliferation; Epithelial Cells; Exposure to; Fibrosis; Flow Cytometry; Functional disorder; Generations; Genes; Genetic Transcription; Goals; Human; Immunology; In Vitro; Inflammation; Inflammatory Response; Injury; Knockout Mice; Knowledge; Laboratories; Lead; Length; Lung; Lung Transplantation; Lung diseases; Malignant Neoplasms; Mediator of activation protein; Mentors; Mesenchymal; Mesenchyme; Modeling; Morphogenesis; Mus; Mutation; Paracrine Communication; Pathogenesis; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Physicians; Play; Population; Prevalence; Prognosis; Protein Secretion; Proteins; Proteomics; Pulmonary Fibrosis; Regenerative capacity; Research; Resource Allocation; Risk Factors; Role; Scientist; Signal Pathway; Signal Transduction; Source; Stromal Cell-Derived Factor 1; Telomere Maintenance; Telomere Maintenance Gene; Testing; Therapeutic; Time; Tissues; Training; Transplantation; United States; Universities; alveolar epithelium; autocrine; base; career; cell type; chemokine; epithelial stem cell; genome editing; idiopathic pulmonary fibrosis; improved; in vivo; migration; mouse model; novel; novel therapeutics; paracrine; prevent; pulmonary function; pulmonary function decline; response; senescence; skills; telomere; theories

Project Summary/Abstract: Idiopathic pulmonary fibrosis (IPF) is a progressive, fibrosing lung disease for which there is no cure. IPF is associated with aging and will likely become more common as our population ages. The pathophysiology of IPF remains incompletely understood, but the alveolar epithelium, specifically the senescent alveolar epithelium, has recently been implicated in this disease. Telomeres are DNA and protein caps on the ends of chromosomes. Short telomeres are a known risk factor for IPF, and mutations in telomere maintenance genes cause IPF. In the setting of telomere dysfunction, alveolar epithelial cells (AECs) rather than dying, preferentially become senescent. Senescent AECs have been shown to secrete pro-survival and pro-fibrotic proteins. This is known as the senescence-associated secretory phenotype (SASP). Multiple SASP proteins have been shown to be capable of inducing their own secretion through autocrine (feed-forward) pathways and to also signal in the traditional paracrine fashion to bystander, non-senescent cells. CXCL12, a SASP protein, has previously been shown to be capable of both autocrine and paracrine signaling in an AEC-like cell line, but the downstream transcriptional and proteomic consequences have not been fully explored. CXCL12 has also been shown to play an important role in lung morphogenesis, but the major cell-type of origin of CXCL12 is not known. The main objective of this proposal is to determine the role of CXCL12 as an autocrine and paracrine mediator originating from a senescent alveolar epithelium and whether loss of CXCL12 signaling from the pulmonary epithelium prevents the fibrotic response. The downstream signals CXCL12 generates on the alveolar epithelium itself will be explored in depth. Aim 1 will utilize a novel conditionally senescent alveolar epithelial-like cell line to explore the consequences of autocrine and paracrine CXCL12 signaling onto the epithelium itself. Aim 2 will seek to determine the function of pulmonary epithelium-derived CXCL12 in a mouse model of pulmonary fibrosis. The knowledge gained from the completion of these studies may promote further research into the role of the alveolar epithelium in IPF and influence the allocation of resources toward the development of CXCL12 pathway-based therapeutics for this disease. Furthermore, this project will provide the applicant the opportunity to develop expertise in lung epithelial cell biology and immunology. The training plan will promote acquisition of advanced laboratory skills including flow cytometry, CRISPR/Cas genome editing, proteomics, and multiple elements of animal modeling. Additionally, didactic courses have been selected to supplement the hands-on training received and promote the advancement of the applicant’s career. Combined with close mentoring and the robust research environment at the University of Pittsburgh, this proposal will support the candidate’s development as an independent physician-scientist.

项目摘要/摘要： 特发性肺纤维化（IPF）是一种进行性纤维化肺部疾病，无法治愈。特发性肺纤维化是 与老龄化有关，并且随着人口老龄化可能会变得更加普遍。病理生理学 IPF 仍不完全清楚，但肺泡上皮，特别是衰老的肺泡 最近发现与这种疾病有关。端粒是 DNA 和蛋白质末端的帽子 染色体。端粒短是 IPF 的已知危险因素，端粒维持基因突变 引起IPF。在端粒功能障碍的情况下，肺泡上皮细胞（AEC）不会死亡， 优先衰老。衰老的 AEC 已被证明能够分泌促生存和促纤维化的物质 蛋白质。这被称为衰老相关分泌表型（SASP）。多种SASP蛋白 已被证明能够通过自分泌（前馈）途径诱导自身分泌， 还以传统的旁分泌方式向旁观者、非衰老细胞发出信号。 CXCL12，一种 SASP 蛋白， 先前已被证明能够在 AEC 样细胞系中同时进行自分泌和旁分泌信号传导，但是 下游转录和​​蛋白质组后果尚未得到充分探索。 CXCL12还具有 已被证明在肺形态发生中发挥重要作用，但 CXCL12 的主要细胞来源类型不是 已知。该提案的主要目标是确定 CXCL12 作为自分泌和旁分泌的作用 源自衰老肺泡上皮的介质以及 CXCL12 信号传导是否丢失 肺上皮可防止纤维化反应。 CXCL12 上生成的下行信号 肺泡上皮本身将被深入探索。目标 1 将利用一种新型条件性衰老肺泡 上皮样细胞系探索自分泌和旁分泌 CXCL12 信号传导对细胞的影响 上皮本身。目标 2 将寻求确定小鼠肺上皮来源的 CXCL12 的功能 肺纤维化模型。从完成这些研究中获得的知识可以进一步促进 研究肺泡上皮在 IPF 中的作用并影响资源分配 开发针对该疾病的基于 CXCL12 通路的疗法。此外，该项目将提供 申请人有机会发展肺上皮细胞生物学和免疫学方面的专业知识。培训计划 将促进获得先进的实验室技能，包括流式细胞术、CRISPR/Cas基因组编辑、 蛋白质组学和动物建模的多个要素。此外，还选择了教学课程 补充所接受的实践培训并促进申请人的职业发展。组合 在匹兹堡大学的密切指导和强大的研究环境下，该提案将 支持候选人作为独立医师科学家的发展。