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，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。目前尚无疗法成功阻止或反向纤维化，这代表了临床的重大需求。纤维化主要发生在软组织（肝脏，肺，肾脏，心脏，皮肤）中 细胞外基质的增殖和沉积。我们最近在肺部以及肝脏中其他的工作表明，细胞外基质僵硬是纤维化的早期和突出事件。至关重要的是，我们和其他人发现，基质从正常水平到纤维水平僵硬，支持成纤维细胞激活到增殖/基质合成状态，以及的影响 矩阵刚度与（和/或添加到）TGF-beta（主要的促纤维化可溶性因子）的影响无关。因此，增加基质刚度会产生一个机械生物学的正反馈回路，从而驱动渐进性纤维化。因此，我们认为应在生理相关的基质刚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措施的新靶标。为了满足这一需求，我们开发了一个细胞培养平台，以研究与肺中新兴纤维化病变相匹配的刚度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，可以在生理相关的机械环境中研究成纤维细胞表型响应，以兼容与高吞吐量，以发现为导向的方法。我们在这里提议在参考肺成纤维细胞系中筛选生物活性分子的库，并测量对关键疾病相关的细胞表型的影响，然后测试候选分子的能力，以改变与IPF和Control Lungs相关的原发性成纤维细胞的纤维源激活，并在所有与型型脂肪中均具有刚度与刚性纤维相配。成功将通过识别具有广泛功能效应的经过验证的命中来定义 调节与疾病相关的原发性人肺成纤维细胞的纤维化激活。识别刚度特异性疗法可以为靶向靶向成纤维细胞的靶向失活提供新的机会，并将田野迈向阻止或逆转进行性纤维化的新方法。