Screening Fibroblast-Matrix Stiffness Interactions to ID New Fibrosis Therapies

描述(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。目前还没有成功阻止或逆转纤维化的治疗方法，这是一个重要的未得到满足的临床需求。纤维化主要发生在软组织(肝、肺、肾、心脏、皮肤)，通过成纤维细胞。 细胞外基质的增殖和沉积。我们最近在肺和肝脏的研究表明，细胞外基质硬化是纤维化的早期和显著事件。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激活到增殖/基质合成状态，以及 基质硬度独立于(和/或增加)转化生长因子-β的影响，转化生长因子-β是主要的促纤维化可溶性因子。因此，增加基质硬度会产生一个机械生物学正反馈环，从而推动进行性纤维化。因此，我们认为应该在生理相关的基质僵硬条件下研究成纤维细胞的行为，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，我们开发了一个细胞培养平台，用于研究与肺内新出现的纤维化病变相匹配的僵硬基质上的成纤维细胞生物学。重要的是，我们的方法首次提供了在与高通量、发现导向的方法兼容的生理相关的机械环境中研究成纤维细胞对分子筛选的表型反应的机会。我们建议在这里筛选生物活性分子文库，并在参考肺成纤维细胞系中测量对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的疾病相关原代成纤维细胞和控制肺的纤维化激活的能力，所有这些都在与新出现的纤维化病变匹配的基质上。成功的定义将是识别在下行中具有广泛功能影响的有效命中 调节与疾病相关的原代人肺成纤维细胞的纤维化激活。僵硬特异性疗法的确定可能为纤维母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。

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.