Screening Fibroblast-Matrix Stiffness Interactions to ID New Fibrosis Therapies

筛选成纤维细胞-基质硬度相互作用以识别新的纤维化疗法

• 批准号: 8445051
• 负责人: Daniel J. Tschumperlin
• 金额: $ 22.71万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-02 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8445051
• 关键词: Address; Behavior; Biological Assay; Biological Models; Biology; Cell Culture Techniques; Cell Line; Cells; Cellular Morphology; Clinical; Collagen; Deposition; Disease; Dissection; Environment; Event; Extracellular Matrix; F-Actin; Feedback; Fibroblasts; Fibronectins; Fibrosis; Hamman-Rich syndrome; Harvest; Heart; Human; Image; Individual; Kidney; Lesion; Libraries; Liver; Lung; Matrix Metalloproteinases; Measures; Mechanics; Molecular; Molecular Bank; Organ; Outcome; Patients; Phenotype; Preclinical Drug Evaluation; Process; Production; Pulmonary Fibrosis; Serum; Skin; Structure of parenchyma of lung; Testing; Therapeutic; Therapeutic Intervention; Tissues; Transforming Growth Factor beta; Work; base; efficacy testing; end stage disease; human TGFB1 protein; in vivo; insight; loss of function; novel strategies; outcome forecast; public health relevance; research study; response; screening; soft tissue; success; therapeutic development

DESCRIPTION (provided by applicant): Fibrosis is pathobiological process common to many tissues and diseases which results in tissue remodeling and loss of function, often necessitating organ replacement or leading to end-stage disease. No therapies are currently available that successfully arrest or reverse fibrosis, and this represents a significant unmet clinical need. Fibrosis occurs predominantly in soft tissues (liver, lung, kidney, heart, skin) through fibroblast proliferation and deposition of extracellular matrix. Our recent work in the lung, and that of others in the liver, demonstrates that extracellular matrix stiffening is an early and prominent event in fibrosis. Critically, we and others have found that matrix stiffening from normal to fibroic levels supports fibroblast activation to a proliferative/matrix synthetic state, and the effects of matrix stiffness are independent of (and/or add to) the effects of TGF-beta, the dominant pro-fibrotic soluble factor. Increasing matrix stiffness thus creates a mechanobiological positive feedback loop that drives progressive fibrosis. We therefore believe fibroblast behaviors should be studied in physiologically relevant matrix stiffness conditions to identify new targets for potential therapeutic intervention relevant to fibrosis. To address this need, we have developed a cell culture platform to study fibroblast biology on matrices of stiffness matched to emerging fibrotic lesions in the lung. Importantly, our approach offers the first opportunity to study fibroblast phenotypic responses to molecular screening within a physiologically relevant mechanical environment compatible with a high throughput, discovery oriented approach. We propose here to screen a library of bioactive molecules and measure effects on key disease-relevant cellular phenotypes in a reference lung fibroblast cell line, and then test candidate molecules for their ability to alter fibrogenic activation of disease relevant primary fibroblasts from IPF and control lungs, all on matrices with stiffness matched to emerging fibrotic lesions. Success will be defined by identification of validated hits with broadly functional effects in down regulating fibrogenic activation of disease-related primary human lung fibroblasts. The identification of stiffness-specific therapies could provide new opportunities for targeted deactivation of fibroblasts and move the field toward new approaches for arresting or reversing progressive fibrosis.

描述（由申请人提供）：纤维化是许多组织和疾病常见的病理生物学过程，其导致组织重塑和功能丧失，通常需要器官置换或导致终末期疾病。目前没有成功阻止或逆转纤维化的疗法，这代表了显著未满足的临床需求。纤维化主要通过成纤维细胞发生在软组织（肝、肺、肾、心脏、皮肤）中。 增殖和细胞外基质沉积。我们最近在肺和其他肝脏的研究表明，细胞外基质硬化是纤维化的早期和突出事件。重要的是，我们和其他人已经发现，从正常到纤维化水平的基质硬化支持成纤维细胞活化到增殖/基质合成状态，并且成纤维细胞活化的作用是促进成纤维细胞的增殖/基质合成。 基质硬度独立于（和/或增加）TGF-β（主要的促纤维化可溶性因子）的作用。因此，增加基质硬度产生了一个机械生物学正反馈回路，驱动进行性纤维化。因此，我们认为应在生理相关基质硬度条件下研究成纤维细胞行为，以确定与纤维化相关的潜在治疗干预的新靶点。为了满足这一需求，我们开发了一种细胞培养平台，以研究与肺中新出现的纤维化病变相匹配的刚度基质上的成纤维细胞生物学。 重要的是，我们的方法提供了第一个机会，研究成纤维细胞表型反应的分子筛选生理相关的机械环境兼容的高通量，发现为导向的方法。我们在此建议筛选生物活性分子库并测量对参考肺成纤维细胞系中关键疾病相关细胞表型的影响，然后测试候选分子改变来自IPF和对照肺的疾病相关原代成纤维细胞的纤维化激活的能力，所有这些都在具有与新出现的纤维化病变匹配的刚度的基质上。成功将通过识别经验证的命中来定义，这些命中具有广泛的功能效应， 调节疾病相关的原代人肺成纤维细胞的纤维化活化。僵硬特异性疗法的鉴定可以为成纤维细胞的靶向失活提供新的机会，并将该领域推向阻止或逆转进行性纤维化的新方法。