Targeting HSC-derived Circulating Fibroblast Precursors in Pulmonary Fibrosis

靶向 HSC 衍生的循环成纤维细胞前体治疗肺纤维化

• 批准号: 8582197
• 负责人: AMANDA C. LARUE
• 金额: --
• 依托单位: RALPH H JOHNSON VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-10-01 至 2017-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8582197
• 关键词: Age; Alveolar; Architecture; Autologous; Automobile Driving; Biological Markers; Blood Circulation; Bone Marrow; Breathing; Cell Transplantation; Cells; Chronic; Chronic lung disease; Clonal Hematopoietic Stem Cell; DDR2 gene; Data; Development; Diagnosis; Disease; Disease Progression; Dust; Effector Cell; Environmental Hazards; Epithelial Cell Proliferation; Evaluation; Exposure to; Extracellular Matrix; FDA approved; Fibroblasts; Fibrosis; Figs - dietary; Flow Cytometry; Gases; Goals; Gulf War; Health; Hematopoietic stem cells; Heterogeneity; Human; In Vitro; Incidence; Lead; Lung; Lung diseases; Methods; Mission; Modeling; Molecular; Mus; Myofibroblast; PTPRC gene; Particulate; Patients; Phenotype; Population; Population Heterogeneity; Pulmonary Fibrosis; Pulse Oximetry; Respiration Disorders; Risk; Role; Silicon Dioxide; Soldier; Staining method; Stains; Symptoms; Testing; Therapeutic; Tissues; Transplantation; Treatment Effectiveness; Veterans; Xenograft Model; alveolar type II cell; base; effective therapy; enhanced green fluorescent protein; environmental chemical; hazard; human disease; improved; in vitro testing; in vivo; insight; monocyte; new therapeutic target; novel; paracrine; particle; peripheral blood; progenitor; public health relevance; reconstitution; response; therapeutic target; trafficking

DESCRIPTION (provided by applicant): Exposure to environmental and chemical hazards is thought to be a major contributing factor to Gulf War illness (GWI) (US Dept VA). Of particular concern for Gulf War Veterans is chronic inhalation of sand, dust and airborne particulates while in theater which pose significant risk for development of respiratory diseases including pulmonary fibrosis (PF). PF is a chronic lung disease characterized by accumulation of extracellular matrix (ECM), destruction of normal lung architecture, and decreased capacity for gas exchange. The activated fibroblast is the primary effector cell in PF. However, targeting activated fibroblasts is challenging due, in part, to the heterogeneity of the population. This heterogeneity is thought to reflect the multiple proposed origins of fibroblasts, making it essential to elucidate the role of fibroblasts from all origins i PF. Using mice whose bone marrow was reconstituted by a clonal population of cells derived from a single enhanced green fluorescent protein positive (EGFP+) hematopoietic stem cell (HSC), we have demonstrated an HSC origin for fibroblasts and activated fibroblasts in multiple tissues. We also identified a CD45+DDR2+ HSC-derived circulating fibroblast precursor (CFP) in peripheral blood of mice and humans that is related to the monocyte, gives rise to activated fibroblasts, produces pro-fibrotic factors, increases with disease, and can be therapeutically targeted. Using a silica model of PF that mimics particulate exposure in Veterans of the Gulf War, we have shown that CFPs increase in circulation with PF and traffic to the fibrotic lung. Together, our findings support the hypothesis that HSC-derived CFPs are critical to progression of PF and can be targeted to inhibit fibrotic progression. This will be tested using our novel clonal HSC cell transplantation method in conjunction with a silica instillation PF model in three Specific Aims. Aim 1 will temporally examine CFP contribution to silica-induced PF and potential of CFPs to serve as an early biomarker for disease and/or response to therapy (Aim 1). Studies in Aim 2 will elucidate mechanisms regulating participation, activation and pro-fibrotic effects of CFPs. Finally, we will examine the effect of therapeutically targeting this unique fibrotic progenitor using both mouse-to-mouse transplant and human-to-mouse xenograft models to demonstrate that inhibition of CFPs can reduce progression of silica-based fibrosis and is applicable to human disease (Aim 3). The proposed studies are paradigm shifting in that most studies focus on targeting the activated fibroblast or relatively mature fibrocyte in fibrosis, whie our data suggests that the more primitive CFP may provide a more effective therapeutic target. These studies are significant in that they are the first to conduct flow cytometric, immunohistochemical, molecular, and functional evaluation of HSC-derived CFPs and their contribution to PF. It is suggested that the number of Veterans diagnosed with Gulf War Illness, including respiratory diseases, is grossly underestimated. As GWI is better defined and the Veteran population serving in the Gulf ages, we expect to see an increased incidence of PF in the Veteran population. Studying early markers of PF is of great relevance to the VA mission as it will allow us to better detect early signs of PF in patients, potentially using CFPs as a biomarker, to dramatically improve effectiveness of treatment prior to loss of normal lung architecture and function. Therefore, these studies have the potential to directly impact Gulf War Veterans' health as well as have far-reaching application to Veterans with multiple types of fibrotic disease.

描述（由申请人提供）： 暴露于环境和化学危害被认为是造成海湾战争疾病（GWI）（美国部VA）的主要因素。海湾战争退伍军人特别关注的是对沙子，灰尘和空中颗粒物的长期吸入，而在剧院中，这对 包括肺纤维化（PF）在内的呼吸道疾病的发展。 PF是一种慢性肺部疾病，其特征是细胞外基质（ECM）的积累，正常肺结构的破坏以及气体交换能力降低。活化的成纤维细胞是PF中的主要效应细胞。但是，靶向活化的成纤维细胞构成挑战，部分原因是人口的异质性。这种异质性被认为反映了成纤维细胞的多重起源，因此阐明所有起源I PF的成纤维细胞的作用至关重要。 使用骨髓的小鼠是由源自单个增强的绿色荧光蛋白阳性（EGFP+）造血干细胞（HSC）的细胞克隆种群重构的，我们已经证明了成纤维细胞的HSC起源和多个组织中活化的成纤维细胞。我们还鉴定了与单核细胞相关的小鼠和人类外周血中的CD45+ DDR2+ HSC衍生的循环成纤维细胞前体（CFP），从而引起活化的成纤维细胞，从而产生促纤维化因子，从而与疾病增加，并可以在治疗方面靶向。使用PF的二氧化硅模型，该模型在海湾战争的退伍军人中模仿颗粒物暴露，我们已经表明，CFP随PF的循环增加而增加，并流动到纤维化肺。总之，我们的发现支持了以下假设：HSC衍生的CFP对PF的进展至关重要，并且可以靶向抑制纤维化进展。这将通过我们的新型克隆HSC细胞移植方法与二氧化硅灌输PF模型结合使用三个特定目的进行测试。 AIM 1将暂时检查CFP对二氧化硅诱导的PF的贡献，以及CFP的潜力作为疾病和/或对治疗反应的早期生物标志物的潜力（AIM 1）。 AIM 2中的研究将阐明调节参与，激活和促纤维化作用的机制 CFP。最后，我们将使用小鼠对小鼠移植和人向小鼠的异种移植模型来检查治疗靶向这种独特的纤维化祖细胞的效果，以证明抑制CFP可以减少基于二氧化硅的纤维化进展，并且适用于人类疾病（AIM 3）。 拟议的研究是范式转移，因为大多数研究都集中在纤维化中靶向活化的成纤维细胞或相对成熟的纤维细胞时，我们的数据表明，更原始的CFP可能提供更有效的治疗靶标。这些研究很重要，因为它们是第一个对HSC衍生的CFP及其对PF的贡献进行流式细胞术，免疫组织化学，分子和功能评估的人。有人认为，诊断出患有海湾战争疾病的退伍军人人数（包括呼吸道疾病）被严重低估了。由于GWI的定义更好，而且在海湾年龄中服务的退伍军人人口，我们预计在退伍军人人口中，PF的发生率会增加。研究PF的早期标记与VA任务非常相关，因为它将使我们能够更好地检测患者的早期PF迹象，并可能将CFP用作生物标志物，以显着提高治疗的有效性，然后再失去正常的肺建筑和功能。因此，这些研究有可能直接影响海湾退伍军人的健康，并对具有多种类型的纤维化疾病的退伍军人进行深远的应用。