Targeting durotaxis in lung injury and fibrosis

靶向肺损伤和纤维化中的杜罗轴

基本信息

批准号：
10532249
负责人：
Benjamin David Medoff
金额：
$ 58.08万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-12-01 至 2025-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10532249
关键词：
Acetylation Acetyltransferase Actins Active Sites Adhesions Affect Area Atomic Force Microscopy Automobile Driving Binding Biochemical Biophysics Breathing Cells Chemotactic Factors Chemotaxis Cicatrix Clathrin-Coated Vesicles Clinical Treatment Clinical Trials Complex Cues Data Development Disease Engineering Enzymes Feedback Fibroblasts Fibrosis Human Hydrogels Image Immune In Vitro Integrin alphaVbeta3 Integrins Knockout Mice Knowledge Laboratories Leukocyte Trafficking Lung Lung diseases Maps Measures Mechanics Mediating Mediator Methods Microtubules Modeling Molecular Mus Myofibroblast PTK2 gene Pathogenesis Pathologic Pathway interactions Patients Pattern Process Pulmonary Fibrosis Recycling Role Signal Transduction Site Slice Spatial Distribution Structure of parenchyma of lung Testing Therapeutic Tissues Treatment Efficacy Tubulin Variant Visualization antifibrotic treatment cell motility cell type chemokine chemokine receptor design experimental study falls fibrotic lung human disease idiopathic pulmonary fibrosis improved in vivo indium-bleomycin inhibitor lung development lung injury mouse model multiphoton microscopy new therapeutic target novel novel therapeutic intervention paxillin recruit response small molecule inhibitor tissue injury

项目摘要

Project Summary/Abstract Idiopathic Pulmonary Fibrosis (IPF) is a fatal lung disease characterized by progressive scarring of the lungs, ultimately impeding the ability to breathe. Pathological recruitment of fibroblasts to sites of tissue injury and subsequent activation into scar-forming myofibroblasts are critical steps in the development and progression of pulmonary fibrosis. Accordingly, the identification of the molecular mediators directing fibroblast recruitment and myofibroblast activation, will not only further enhance our understanding of the pathogenesis of lung fibrosis, but also provide rational therapeutic targets for novel anti-fibrotic therapies. We and others have recently shown that increased matrix stiffness in fibrotic lungs promotes mechano-activation of fibroblasts. Further, matrix stiffness amplifies tissue fibrosis by locking stiffness-activated myofibroblasts on a mechanical positive feedback loop, by not fully understood mechanisms. We have recent evidence that matrix stiffness gradients produced in fibrotic lung tissues promote fibroblast recruitment to sites of tissue injury via durotaxis – the directed migration of cells from regions of lower to higher stiffness, which occurs independently of diffusible chemoattractants or substrate-bound haptotactic cues. As fibroblast “durotax” to regions of increased stiffness, the stiffness of these regions would drive the arriving fibroblasts to differentiate into myofibroblasts. Consequently, the central hypotheses of this proposal are that: (1) fibroblasts are recruited to sites of focal tissue injury via durotaxis, a mechanism in which cells migrate up stiffness gradients independently of chemotactic signals; and (2) that inhibition of fibroblast durotaxis has the potential to be a new therapeutic strategy for IPF. The studies proposed in this application are designed to visualize fibroblast durotaxis in vivo, to define molecular mechanisms of fibroblast durotaxis and to develop novel therapeutic strategies to inhibit durotaxis. Specifically, we propose: (1) To image fibroblast durotaxis ex vivo using multiphoton microscopy in precision cut lung slices and to investigate the therapeutic efficacy of targeting fibroblast durotaxis in vivo in a mouse model of lung fibrosis by inhibiting the mechanosensitive FAK/Paxillin pathway; (2) To define mechanisms by which the αvβ3/FAK/Paxillin pathway regulates matrix rigidity sensing and durotaxis in IPF fibroblasts. We will investigate both biochemical and biophysical regulators of αvβ3 integrin and their role in fibroblast durotaxis; and (3) To define mechanisms by which actin-microtubule crosstalk mediates fibroblast durotaxis. Specifically, we will investigate mechanisms by which α-TAT1-mediated microtubule acetylation controls dynamic recycling of αvβ3/FAK/Paxillin complexes in durotactic cells. We will also test the role of αTAT-1 in fibroblast durotaxis and pulmonary fibrosis in vivo in the bleomycin model of lung fibrosis, using fibroblasts-specific αTAT-1 KO mice generated by our laboratory. The experiments proposed in this application will provide new knowledge regarding the role of fibroblast durotaxis in the development and progression of lung fibrosis, and the potential efficacy of anti-durotactic therapy for the treatment of lung fibrosis.

项目概要/摘要特发性肺纤维化（IPF）是一种致命的肺部疾病，其特征是肺部进行性疤痕形成，最终阻碍成纤维细胞向组织损伤部位的病理性募集。随后激活形成疤痕的肌成纤维细胞是肌成纤维细胞发育和进展的关键步骤因此，确定了指导成纤维细胞募集的分子介质。和肌成纤维细胞的激活，不仅将进一步加深我们对肺病发病机制的认识我们和其他人已经为新型抗纤维化疗法提供了合理的治疗靶点。最近表明，纤维化肺中基质硬度的增加可促进成纤维细胞的机械激活。此外，基质硬度通过将硬度激活的肌成纤维细胞锁定在机械上来放大组织纤维化。正反馈循环，通过尚未完全理解的机制我们最近有证据表明基质刚度。纤维化肺组织中产生的梯度通过杜罗轴促进成纤维细胞募集到组织损伤部位 – 细胞从较低硬度区域向较高硬度区域的定向迁移，其发生与扩散无关作为成纤维细胞“durotax”到硬度增加的区域，这些区域的硬度将驱动到达的成纤维细胞分化为肌成纤维细胞。 (1) 成纤维细胞被募集到病灶部位通过杜罗轴（durotaxis）造成的组织损伤，这是一种细胞独立于硬度梯度向上迁移的机制趋化信号；(2) 抑制成纤维细胞杜罗趋化有可能成为一种新的治疗方法本申请中提出的研究旨在可视化体内成纤维细胞的 durotaxis，定义成纤维细胞硬度的分子机制并开发新的治疗策略来抑制具体来说，我们建议：（1）使用多光子显微镜对成纤维细胞的 durotaxis 进行离体成像。精密切割肺切片并研究体内靶向成纤维细胞杜罗塔斯的治疗效果通过抑制机械敏感性 FAK/Paxillin 通路建立小鼠肺纤维化模型 (2) 定义 αvβ3/FAK/Paxillin 通路调节 IPF 中基质刚性感应和 durotaxis 的机制我们将研究 αvβ3 整合素的生化和生物物理调节因子及其在成纤维细胞中的作用。 (3) 定义肌动蛋白-微管串扰介导成纤维细胞的机制具体来说，我们将研究 α-TAT1 介导的微管乙酰化机制。控制 αvβ3/FAK/Paxillin 复合物在杜罗定向细胞中的动态回收。 αTAT-1 在博来霉素肺纤维化模型中体内成纤维细胞杜罗轴性和肺纤维化中的作用，使用本申请中提出的实验是由我们实验室生成的成纤维细胞特异性αTAT-1 KO小鼠。将提供有关成纤维细胞 durotaxis 在发育和进展中的作用的新知识肺纤维化，以及抗杜罗定向疗法治疗肺纤维化的潜在疗效。