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.

项目摘要/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，最终阻碍了呼吸的能力。将成纤维细胞的病理募集到组织损伤部位和随后激活疤痕形成肌纤维细胞是开发和发展的关键步骤肺纤维化。彼此之间，指导成纤维细胞募集的分子介质的鉴定肌纤维细胞激活不仅会进一步增强我们对肺发病机理的理解纤维化，但也为新型抗纤维化疗法提供了合理的治疗靶标。我们和其他人有最近表明，纤维化肺的基质刚度增加会促进成纤维细胞的机械激活。此外，矩阵刚度放大器组织纤维化通过将刚度激活的肌纤维细胞锁定在机械上积极的反馈循环，不完全了解机制。我们最近有证据表明矩阵刚度在纤维化肺正时产生的梯度可通过Durotaxis促进组织损伤部位的成纤维细胞募集 - 细胞从较低刚度到更高刚度的区域的定向迁移，这独立于扩散化学吸引剂或底物结合的触觉提示。作为成纤维细胞“ Durotax”到刚度增加的区域这些区域的刚度将驱动到达的成纤维细胞分化为肌纤维细胞。因此，该提案的中心假设是：（1）招募成纤维细胞到焦点部位通过Durotaxis进行组织损伤，这种机制在其中细胞迁移刚度梯度独立于趋化信号；（2）抑制成纤维细胞durotaxis有可能成为一种新疗法 IPF的策略。本应用中提出的研究旨在可视化体内成纤维细胞durotaxis，定义成纤维细胞Durotaxis的分子机制，并开发新的治疗策略以抑制 durotaxis。特别是，我们提出：（1）使用多光子显微镜在体内图像成纤维细胞durotaxis ex extime 精确切割肺切片并研究靶向成纤维细胞Durotaxis体内的治疗效率肺纤维化小鼠模型通过抑制机械敏感的FAK/Paxillin途径；（2）定义 αVβ3/fak/paxillin途径调节矩阵刚度敏感和durotaxis的机制成纤维细胞。我们将研究αVβ3整合素的生化和生物物理调节剂及其在成纤维细胞durotaxis; （3）定义肌动蛋白 - 微动物串扰的机制 durotaxis。特别是，我们将研究α-TAT1介导的微管乙酰化的机制控制Durotactic细胞中αVβ3/FAK/Paxillin复合物的动态回收。我们还将测试肺纤维素模型中的成纤维细胞Durotaxis和肺纤维化中的αTAT-1，使用肺纤维化模型，使用使用我们实验室产生的成纤维细胞特异性αTAT-1 KO小鼠。本应用程序中提出的实验将提供有关成纤维细胞Durotaxis在发展和发展中的作用的新知识肺纤维化，以及抗尿路疗法对肺纤维化治疗的潜在有效性。