Neuroimmune Molecules in Scleroderma Lung Fibrosis

硬皮病肺纤维化中的神经免疫分子

• 批准号: 9233186
• 负责人: Erica L Herzog
• 金额: $ 41.82万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9233186
• 关键词: Affect; Appearance; Area; Autoimmune Diseases; Binding; Biochemical; Biological Assay; Biomedical Engineering; Bleomycin; Blood; Bone Marrow; Bone Marrow Transplantation; Breathing; Cells; Characteristics; Chimera organism; Clinical; Collagen; Cutaneous; Cytometry; DCC gene; Dependence; Development; Disease; Doxycycline; Experimental Models; Extracellular Matrix; Fibrosis; Genes; Goals; Histologic; Human; Immune; Immunologics; Inflammatory; Interstitial Lung Diseases; Knockout Mice; Laboratories; Laminin; Leukocytes; Light; Longitudinal cohort; Lung; Mechanics; Mediating; Mesenchymal; Modeling; Mus; Mutation; NTN1 gene; Neuroimmune; Neurons; PTPRC gene; Pathogenesis; Pathway interactions; Patient-Focused Outcomes; Patients; Phenotype; Polymers; Population; Production; Property; Proteins; Publishing; Pulmonary Fibrosis; Reporting; Role; Scleroderma; Site; Structure of parenchyma of lung; Systemic Scleroderma; Techniques; Testing; Tissues; Viral; Visceral; Work; base; combinatorial; immunoregulation; improved; indium-bleomycin; injury and repair; insight; monocyte; mouse model; neuronal guidance; neutralizing antibody; novel; novel therapeutics; null mutation; overexpression; public health relevance; receptor; repaired; response; therapeutic target

 DESCRIPTION (provided by applicant): Scleroderma (Systemic Sclerosis, SSc) is an idiopathic autoimmune disease characterized by cutaneous and visceral fibrosis in which many patients are affected by interstitial lung disease (SSc-ILD) which lacks specific, highly efficacious therapy. The response of SSc-ILD to immunomodulatory agents and in some cases bone marrow transplantation suggests involvement of leukocytes in disease pathogenesis. Detailed examination of lung tissue obtained from patients with SSc-ILD frequently reveals inflammatory cells juxtaposed with normal and fibrotic extracellular matrix (ECM). It is therefore notable that fibrocytes, a population of leukocytes displaying mesenchymal characteristics, have in several studies demonstrated enhanced accumulation in the SSc-ILD blood and/or lung. The significance of these cells and the factors regulating their appearance in this clinical context remains unknown. Published and preliminary work from our laboratory indicates the laminin-like neuroimmune molecule Netrin-1 (NTN-1) is important in this setting. NTN-1 stimulates cellular attraction via binding to its attractive receptor, Deleted in Colorectal Cancer-1 (DCC-1) while cellular repulsion and invasion is driven by interactions with its repulsive receptor, Uncoordinated-5a (UNC5a). We explored the relevance of these pathways to SSc-ILD using a novel translational platform based on decellularized human lungs to demonstrate that the accumulation of collagen-producing leukocytes is regulated by biochemical and mechanical aspects of the human lung ECM, and that SSc-ILD PBMCs produce more fibrocytes when exposed to the digestible components of the fibrotic Scleroderma lung. NTN-1 expression is increased on CD14lo monocytes in SSc-ILD where it regulates leukocyte-matrix interactions and TGF-β1 secretion by PBMCs. Last, studies performed in the bleomycin model of pulmonary fibrosis demonstrate that partial deficiency of NTN-1 ameliorates histologic and biochemical readouts of fibrosis, TGF-β1 production, and the accumulation of CD45+ColIα1+ cells, indicating that NTN-1 neutralization might be a therapeutic target in this context. Exploration of these areas has the potential to advance the understanding of lung involvement in SSc. Therefore, in Aim 1, we will use standard immunolocalization techniques and multiparametric mass cytometry to define the expression of NTN-1, DCC-1, and UNC5a on innate and adaptive immune cells in the lungs and blood of two well characterized longitudinal cohort of subjects with SSc-ILD. In Aim 2, we use neutralizing antibodies and virally mediated overexpression strategies combined with mechanically tunable polymer networks rendered bioactive through conjugation with normal and SSc-ILD lung matrix to determine whether Netrin-1's modulation of fibrocyte accumulation results from receptor mediated interactions with ECM components or from the combinatorial influence of the protein composition and stiffness of the underlying substrate. In Aim 3, we use mice with null mutations of UNC5a and DCC-1, as well as mice with cell specific deletion of NTN-1 and TGF-β1 to determine their contribution to fibrosis and intrapulmonary fibrocyte accumulation seen in two separate models of experimentally-induced lung fibrosis.

 描述（由申请人提供）：硬皮病（系统性硬化症，SSc）是一种特发性自身免疫性疾病，其特征是皮肤和内脏纤维化，其中许多患者患有间质性肺病（SSc-ILD），缺乏特异性、高效的治疗方法。 SSc-ILD 对免疫调节剂的影响以及在某些情况下的骨髓移植表明白细胞参与疾病发病机制。对 SSc-ILD 患者肺组织的详细检查经常会发现炎症细胞与正常和纤维化细胞外基质 (ECM) 并存。因此，值得注意的是，纤维细胞（表现出间充质特征的白细胞群）在多项研究中表明，纤维细胞在细胞外基质中的积累增强。 SSc-ILD 血液和/或肺中这些细胞的重要性以及在这种临床背景下调节其外观的因素仍然未知。我们实验室已发表的初步工作表明层粘连蛋白样。神经免疫分子 Netrin-1 (NTN-1) 在这种情况下很重要，NTN-1 通过与其吸引受体结合来刺激细胞吸引力，在结直肠癌-1 (DCC-1) 中被删除，而细胞排斥和侵袭则由与它的相互作用驱动。我们使用基于去细胞人肺的新型翻译平台探索了这些途径与 SSc-ILD 的相关性，以证明胶原蛋白生成的积累。白细胞受人肺 ECM 的生化和机械方面的调节，并且当 SSc-ILD PBMC 暴露于纤维化硬皮病肺的可消化成分时，SSc-ILD 中 CD14lo 单核细胞上的 NTN-1 表达增加。最后，在博莱霉素肺纤维化模型中进行的研究表明，调节白细胞-基质相互作用和 PBMC 分泌 TGF-β1。 NTN-1 的部分缺乏改善了纤维化、TGF-β1 产生和 CD45+ColIα1+ 细胞积累的组织学和生化读数，表明 NTN-1 中和可能是在这种情况下的治疗靶点。因此，在目标 1 中，我们将使用标准免疫定位技术和多参数质谱流式技术来定义 NTN-1 的表达， DCC-1 和 UNC5a 对两个特征明确的 SSc-ILD 受试者的肺部和血液中的先天性和适应性免疫细胞的影响在目标 2 中，我们使用中和抗体和病毒介导的过表达策略，并结合机械可调的聚合物网络。通过与正常和 SSc-ILD 肺基质结合而具有生物活性，以确定 Netrin-1 对纤维细胞积累的调节是否是由受体介导的与 ECM 成分的相互作用引起的，还是来自以下因素的组合影响：在目标 3 中，我们使用 UNC5a 和 DCC-1 无效突变的小鼠以及 NTN-1 和 TGF-β1 细胞特异性缺失的小鼠来确定它们对纤维化和纤维化的贡献。在两个不同的实验诱导的肺纤维化模型中观察到肺内纤维细胞积聚。