Novel Mechanisms of Pulmonary Fibrosis

肺纤维化的新机制

基本信息

批准号：
10660225
负责人：
Francis Xavier McCormack
金额：
$ 64.31万
依托单位：
UNIVERSITY OF CINCINNATI
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-05-20 至 2027-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10660225
关键词：
Acceleration Acids Actins Address Alveolar Alveolar Macrophages Animals Asbestos Attenuated Automobile Driving Biochemical Bone Marrow Bone Matrix Bone Resorption Calcitonin Receptor Calcium Cell Differentiation process Cell Lineage Cell physiology Cells Clinical Clinical Trials Coculture Techniques Complex Defect Development Disease Enzymes Epithelium Exposure to Extracellular Matrix FDA approved Fibroblasts Fibrosis Gene Expression Gene Expression Profile Genes Genetic Transcription Giant Cells Goals Histologic Homeostasis Host Defense Human Hydrochloric Acid Hydroxyapatites In Vitro Inflammasome Inhalation Injury Learning Lesion Liquid substance Lung Lung diseases Macrophage Macrophage Colony-Stimulating Factor Measures Mediating Minerals Molecular Target Mus Mutation Mycobacterium tuberculosis Myelogenous Myeloid Cells Occupational Osteoclasts PPAR gamma Particulate Pathogenesis Pathway interactions Patients Pattern Peptide Hydrolases Phospholipid Metabolism Physiological Planet Earth Pneumoconiosis Predisposition Prevalence Process Production Proliferating Proteins Proton Pump Pulmonary Alveolar Proteinosis Pulmonary Challenge Pulmonary Fibrosis Pulmonary alveolar microlithiasis Reporting Role Silicon Dioxide Silicosis Slice Specific qualifier value Stereotyping Structure of parenchyma of lung Syndrome TNFSF11 gene Testing Tissues Transforming Growth Factor beta Up-Regulation Zoledronic Acid autosome bisphosphonate black lung bone calcium phosphate cathepsin K cell injury cell transformation cytokine experimental study fibrotic lung fibrotic lung disease gene repression human RNA sequencing in vivo inorganic phosphate insight lipid metabolism migration monocyte mouse model novel osteoclastogenesis particle pharmacologic pre-clinical prevent programs protein expression recruit response single-cell RNA sequencing sodium-phosphate cotransporter proteins surfactant tartrate-resistant acid phosphatase theories

项目摘要

This proposal explores the role of a pulmonary osteoclast-like cell (POLC) in silicosis. We first discovered POLCs while studying pulmonary alveolar microlithiasis (PAM), a rare, autosomal recessive disorder caused by mutations in the epithelial sodium phosphate co-transporter, Npt2b. Phosphate accumulates in the alveolar lining fluid and complexes with calcium to form spherical hydroxyapatite microliths containing bone matrix proteins and surfactant components. Contact of microliths with tissue resident alveolar macrophages (TR-AM) and recruited monocytes (Mo-AM) induces osteoclastic transformation in the Npt2b-/- mouse, with expression of the full repertoire of osteoclast signature gene and protein expression in multinucleated giant cells (MNGC) including tartrate resistant acid phosphatase (TRAP), cathepsin K (CTSK), calcitonin receptor (CALCR) and the proton pump, ATP6V0D2. Single cell RNA sequencing (scRNAseq) of human PAM lung also confirmed a robust osteoclast signature in AM, and IHC confirmed the presence of TRAP and CTSK positive MNGC. Like humans, Npt2b-/- animals develop modest pulmonary fibrosis and a marked restrictive physiologic defect. We found that microliths induce alveolar expression of the requisite osteoclastogenic cytokines for POLC differentiation (i.e. M- CSF, RANKL) and expression of hydrochloric acid and CTSK that both dissolve stones and damage tissues. We were surprised to find that the Npt2b-/- mice also develop pulmonary alveolar proteinosis, which has not been reported in PAM, but is known to be associated with another particulate exposure syndrome, silicoproteinosis. This led us to consider that osteoclastic transformation of TR-AM and Mo-AM is a stereotypic response to inhaled particles, resulting in the concomitant loss of AM surfactant catabolic and host defense functions (in the form of susceptibility to Tb). Indeed, snRNAseq, rtPCR measures of gene expression and IHC assessments revealed that silica challenge is also associated with RANKL-dependent osteoclastic transformation of BAL and parenchymal cells, and TRAP, CTSK and hydrochloric acid production, culminating in destructive remodeling and pulmonary fibrosis, and associated with a decrease in proteins required for lipid metabolism. Anti- RANKL treatment of mice attenuates the silicoproteinosis, POLC formation, fibrosis and the restrictive physiologic defect that occurs in silica-challenged mice. The differential tissue responses to dissolvable vs. persistent particulates forms the basis for our hypothesis that persistent acid and matrix degrading enzymes produced by POLC may be primary drivers of particulate-induced fibrosis. To test this hypothesis in three aims, we will determine; 1) Cell lineages and alveolar factors that give rise to POLC; 2) The role of osteoclastic functions and products in silica- induced fibrosis; and 3) The role of POLC in the pathogenesis of pulmonary fibrosis, in vivo. This proposal directly addresses mechanisms relevant to the alarming rise in the prevalence of progressive massive fibrosis in black lung claimants, and successful completion of the aims will provide preclinical evidence supporting anti- osteoclastic strategies such as bisphosphonates and anti-RANKL in exposed subjects.

该提案探讨了肺破骨细胞样细胞（POLC）在硅化作用中的作用。我们首先发现了Polcs 同时研究肺肺泡微石症（PAM），这是一种罕见的常染色体隐性疾病上皮磷酸钠共转运蛋白的突变，NPT2B。磷酸盐积聚在肺泡内膜中用钙的液体和络合物形成含有骨基质蛋白和的球形羟基磷灰石微石表面活性剂成分。微石与组织驻留的肺泡巨噬细胞（TR-AM）接触并招募单核细胞（MO-AM）在NPT2B - / - 小鼠中诱导破骨碎屑转化，并表达完整多核巨细胞（MNGC）中破骨细胞特征基因和蛋白表达的曲目包括防strate抗性酸性磷酸酶（陷阱），组织蛋白酶K（CTSK），降钙素受体（CALCR）和质子泵，ATP6V0D2。人PAM肺的单细胞RNA测序（SCRNASEQ）也证实了坚固的 AM中的破骨细胞签名，IHC证实了陷阱和CTSK阳性MNGC的存在。像人类一样 NPT2B - / - 动物会出现适度的肺纤维化和明显的限制性生理缺陷。我们发现微石诱导必要的破骨细胞因子的肺泡表达进行POLC分化（即M- CSF，RANKL）和盐酸和CTSK的表达既可以溶解石头和损坏组织。我们惊讶地发现NPT2B - / - 小鼠也会出现肺肺泡蛋白质病，这还不是在PAM中报道，但已知与另一种颗粒状暴露综合征，有硅蛋白酶病有关。这导致我们认为Tr-AM和Mo-AM的破骨碎屑转化是对吸入的刻板印象的反应颗粒，导致AM表面活性剂分解代谢和宿主防御功能的损失（以对结核病的敏感性）。实际上，SNRNASEQ，基因表达和IHC评估的RTPCR测量二氧化硅挑战还与BAL和实质细胞，陷阱，CTSK和盐酸产生，最终导致破坏性重塑和肺纤维化，并与脂质代谢所需的蛋白质减少有关。反兰克小鼠的治疗减弱了硅蛋白质病，POLC形成，纤维化和限制性生理缺陷这发生在二氧化硅挑战的小鼠中。对溶解的差异组织反应与持续的颗粒构成了我们假设的基础，即POLC生产的持续酸和基质降解酶是颗粒引起的纤维化的主要驱动因素。为了在三个目标中检验这一假设，我们将确定； 1）细胞产生POLC的谱系和牙槽因子； 2）整骨功能和产物在二氧化硅中的作用诱导纤维化； 3）POLC在体内的肺纤维化发病机理中的作用。该建议直接解决了与黑色进行性大规模纤维化患病率的令人震惊的上升相关的机制肺索赔人以及成功完成目标将提供支持反对的临床前证据裸露受试者中的破骨策略，例如双膦酸盐和抗裂痕。