Inhibition of Fgr Prevents Pulmonary Fibrosis

抑制 Fgr 可预防肺纤维化

基本信息

批准号：
10901018
负责人：
Amitava Mukherjee
金额：
$ 32.6万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
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：
2023-09-15 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10901018
关键词：
Alveolar Cell Biological Markers Biological Models Bone Marrow COVID-19 patient Cell Aging Cell Cycle Cell Senescence Induction Cell Separation Cells Chest Clinical Trials Cyclin-Dependent Kinase Inhibitor 2A Data Dendritic Cells Development Disease Enzymes Exposure to Fibrosis Genes Goals Human In Vitro Inflammatory Knockout Mice Longevity Lung Macrophage Mediating Molecular Mus Oxidative Stress Patients Pharmaceutical Preparations Phenotype Physiological Processes Play Poison Prevention Prevention approach Property Protein Tyrosine Kinase Proteins Pulmonary Fibrosis Radiation Reactive Oxygen Species Reporting Role SARS-CoV-2 infection Silicon Dioxide Silicosis Therapeutic Up-Regulation Wild Type Mouse beta-Galactosidase chemokine cytokine fibrotic lung idiopathic pulmonary fibrosis improved in vivo inhibitor innovation ionization irradiation knock-down migration monocyte mouse model neutrophil novel strategies pharmacologic prevent profibrotic cytokine recruit senescence single-cell RNA sequencing small hairpin RNA small molecule therapeutically effective transcriptome sequencing

项目摘要

Pulmonary fibrosis can arise spontaneously as in idiopathic pulmonary fibrosis (IPF), or following ionizing irradiation, silica exposure, or COVID-19 infection. Senescence has been implicated in both radiation-induced pulmonary fibrosis (RIPF) and IPF. Senescence precedes fibrosis in mouse RIPF and reported in COVID-19 patients. Clearance of senescent cells (SCs) by senolytic drugs present an attractive anti-fibrosis strategy. However, senolytics may be non-specific and remove potentially beneficial as well as harmful SCs. Little is known about how senescent cells induce pulmonary fibrosis. SCs exit cell cycle, induce p16, increase senescence associated beta-galactosidase (SA-𝝱-gal), and secrete senescence associated secretory phenotype (SASP) proteins. SASP include proinflammatory cytokines that may initiate and exacerbate fibrosis. Therefore, using radiation as a model system to induce senescence, we propose to establish and validate a specific mitigator of lung fibrosis by targeting one senescent protein while not removing the SCs. By RNA-seq of pure SCs and by single cell RNA-seq (scRNAseq) of irradiated lungs, we discovered that tyrosine kinase Fgr is highly induced in senescent cells. Fgr positive mouse lung cells induce biomarkers of fibrosis in vitro, and Fgr inhibitor, TL02-59 treatment, abrogates the induction of profibrotic genes. Our preliminary data show: 1) in mouse lung senescent cells, Fgr inhibitor TL02-59 abrogates the induction of profibrotic genes in target cells, 2) in lung cells from Fgr knockout mouse, senescence occurs with radiation, but fail to induce profibrotic genes in target cells, 3) human lungs from RIPF, silicosis, and IPF show induction of Fgr and senescent marker p16, 4) TL02-59 reduces secretion of profibrotic SASP chemokines for attracting immunocytes, 5) inhibition of Fgr in vivo, either by TL02-59, or by senescence inhibitor MMS350, reduces expression of fibrotic genes. We hypothesize, Fgr in senescent cells initiates pulmonary fibrosis by secretion of proinflammatory SASP proteins and by recruiting inflammatory immunocytes to the lungs. We further hypothesize that inhibition of Fgr by small molecule TL02-59 or indirectly by the senescence inhibitor, MMS350, will prevent lung fibrosis. We will use mouse primary lung cells from C57BL/6 control and Fgr knockout (Fgr-/-) mice to confirm the role of Fgr in vivo in C57BL/6 and C57BL/6 Fgr-/- mice. Finally, we will establish that specific inhibition of Fgr prevents lung fibrosis. Specific Aim 1 will establish that lung cell senescence induces Fgr and initiates pulmonary fibrosis by the actions of secreted proinflammatory SASP and recruitment to the lungs of bone marrow derived inflammatory cells. Specific Aim 2 will establish that Fgr is required for pulmonary fibrosis using Fgr -/- mice. Specific Aim 3 will establish that specific and targeted pharmacologic inhibition of Fgr blocks lung fibrosis. Our innovative approach will establish that Fgr induction in senescent lung cells causes pulmonary fibrosis through secretion of proinflammatory SASP proteins and chemotactic recruitment of inflammatory cells. Our proposed studies will lead to the development of a new mitigator of pulmonary fibrosis.

肺纤维化可自发发生，如特发性肺纤维化 (IPF)，或电离后发生辐射、二氧化硅暴露或 COVID-19 感染都与辐射引起的衰老有关。肺纤维化 (RIPF) 和 IPF 小鼠 RIPF 中的衰老先于纤维化，并在 COVID-19 中有所报道。通过衰老药物清除衰老细胞（SC）是一种有吸引力的抗纤维化策略。然而，senolytics 可能是非特异性的，并且几乎无法消除潜在有益和有害的 SC。已知衰老细胞如何诱导肺纤维化 SC 退出细胞周期、诱导 p16 增加。衰老相关的β-半乳糖苷酶（SA--gal），并分泌衰老相关的分泌物表型 (SASP) 蛋白 SASP 包括可能引发和加剧纤维化的促炎细胞因子。因此，利用辐射作为诱导衰老的模型系统，我们建议建立并验证一个通过靶向一种衰老蛋白而不去除 SC 来特异性缓解肺纤维化。通过纯 SC 和受辐射肺部的单细胞 RNA 测序 (scRNAseq)，我们发现酪氨酸激酶 Fgr 是 Fgr 阳性小鼠肺细胞在体外诱导纤维化生物标志物。抑制剂，TL02-59 治疗，消除了促纤维化基因的诱导：1) 在。小鼠肺衰老细胞，Fgr 抑制剂 TL02-59 消除靶细胞中促纤维化基因的诱导，2) 在 Fgr 敲除小鼠的肺细胞中，辐射会导致细胞衰老，但不能诱导促纤维化基因靶细胞，3) 来自 RIPF、硅肺病和 IPF 的人肺显示 Fgr 和衰老标记物的诱导 p16, 4) TL02-59 减少促纤维化 SASP 趋化因子的分泌，以吸引免疫细胞，5) 抑制 Fgr 在体内通过 TL02-59 或衰老抑制剂 MMS350 降低纤维化基因的表达。衰老细胞中的 Fgr 通过分泌促炎性 SASP 蛋白引发肺纤维化并通过将炎症免疫细胞募集到肺部，我们进一步研究了小分子对 Fgr 的抑制作用。我们将使用TL02-59分子或衰老抑制剂MMS350间接预防肺纤维化。来自 C57BL/6 对照和 Fgr 敲除 (Fgr-/-) 小鼠的小鼠原代肺细胞，以确认 Fgr 在体内的作用最后，我们将确定 Fgr 的特异性抑制可预防肺损伤。具体目标 1 将确定肺细胞衰老诱导 Fgr 并引发肺纤维化。通过分泌促炎性 SASP 的作用并将骨髓募集至肺部具体目标 2 将使用 Fgr -/- 确定 Fgr 是肺纤维化所必需的。具体目标 3 将确定 Fgr 的特异性和靶向药理抑制可阻止肺纤维化。我们的创新方法将确定衰老肺细胞中的 Fgr 诱导通过促炎性 SASP 蛋白的分泌和趋化性募集引起肺纤维化我们提出的研究将导致开发一种新的肺纤维化缓解剂。