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.

肺纤维化可以像特发性肺纤维化（IPF）一样发起，或者在电离之后辐照，二氧化硅暴露或共vid-19感染。在两个辐射诱导的肺纤维化（RIPF）和IPF。衰老先于小鼠RIPF的纤维化，并在Covid-19中报告患者。通过鼻溶剂清除感觉细胞（SC）提出了一种有吸引力的抗纤维化策略。但是，塞溶剂剂可能是非特异性的，并且可以消除潜在的有益和有害的SC。几乎没有了解感觉细胞如何诱导肺纤维化。 SCS出口细胞周期，诱导P16，增加感应相关的β-半乳糖苷酶（SA-𝝱-GAL）和秘密相关的秘密表型（SASP）蛋白质。 SASP包括可能启动和加剧纤维化的促炎细胞因子。因此，使用辐射作为模型系统诱导感应，我们建议建立和验证A 通过靶向一种感觉蛋白，同时不去除SC，对肺纤维化的特异性缓解剂。由RNA-seq的纯SC和受辐照肺的单细胞RNA-Seq（Scrnaseq），我们发现酪氨酸激酶FGR为在感觉细胞中高度诱导。 FGR阳性小鼠肺细胞在体外诱导纤维化生物标志物，FGR诱导抑制剂TL02-59处理废除了纤维化基因的诱导。我们的初步数据显示：1）小鼠肺感觉细胞FGR抑制剂TL02-59消除了靶细胞中纤维化基因的诱导，2）在FGR基因敲除小鼠的肺细胞中，辐射时会发生感应，但无法诱导纤维化基因靶细胞，3）RIPF的人肺，硅和IPF显示FGR和感觉标记的诱导 P16，4）TL02-59减少了吸引免疫细胞的纤维化SASP趋化因子的分泌，5）抑制由TL02-59或通过感应抑制剂MMS350在体内的FGR可以降低纤维化基因的表达。我们假设，感觉细胞中的FGR通过促炎SASP蛋白的分泌引发肺纤维化并通过将炎症性免疫细胞募集到肺部。我们进一步假设小型抑制FGR 分子TL02-59或通过感应抑制剂MMS350间接地防止肺纤维化。我们将使用来自C57BL/6对照和FGR敲除（FGR - / - ）小鼠的小鼠原代肺细胞以确认FGR在体内的作用在C57BL/6和C57BL/6 FGR - / - 小鼠中。最后，我们将确定对FGR的特定抑制可防止肺纤维化。特定的目标1将确定肺部细胞感应会诱导FGR并启动肺纤维化通过分泌的促炎SASP和向骨髓肺招募的作用衍生的炎性细胞。特定的目标2将确定使用FGR的肺纤维化需要FGR - / - 老鼠。具体目标3将确定FGR阻滞肺纤维化的特定和靶向药物术抑制。我们的创新方法将确定感觉肺细胞中的FGR诱导通过分泌促炎SASP蛋白和趋化性募集，引起肺纤维化炎性细胞。我们提出的研究将导致开发新的肺纤维化缓解剂。