Fibrotic microtissue chips for screening of anti-fibrotic therapies

 描述：纤维化，如终末期特发性肺纤维化(IPF)、心力衰竭、肝硬变和肾脏疾病，可导致器官衰竭，目前尚无治愈方法。已经确定了抗纤维化治疗的候选药物；然而，由于疾病进展缓慢和临床试验相关的高昂成本，这些实验室发现阻碍了将其转化为临床实践。为了在长期和昂贵的临床试验中证明新疗法的开发是合理的，需要一个能够提供抗纤维化疗法有效性的早期证据的体外筛选平台。然而，现有的 体外模型，以全面概括纤维化疾病的生理病理特征， 如肺纤维化时用力肺活量(FVC)减少和主要细胞外基质(ECM)重构，已显著延缓了急需的抗纤维化治疗的发展。本项目的目标是研制一种微生理性肺纤维化。 微组织芯片“设备，可以实现抗纤维化治疗的早期筛选。我们最近采用微制造技术在聚二甲基硅氧烷(PDMS)微孔阵列中组装了3D成纤维细胞填充的亚毫米微组织。在每个微孔中，成纤维细胞自发收缩和组装基质蛋白，如胶原蛋白，形成固定在一对悬臂之间的排列好的微组织。我们最近的初步数据表明，由于边界条件引导细胞和细胞外基质的自组装，在微组织中可以形成不同的上皮和细胞外基质层。在适当的生化和机械刺激下，可以在这些微组织中进一步诱导驻留细胞的纤维分化。在这个项目中，我们将全面描述诱导微组织的纤维化特性，如模拟强制通风下的顺应性和细胞外基质的沉积。然后，我们将用FDA最近批准的两种治疗IPF的药物来测试它的有效性。这项应用的具体目的是：1)制造微组织阵列并诱导纤维化过渡；2)校准纤维化微组织的力学和组织学特性，并使用一小套训练型抗纤维化药物来优化系统；以及3)评估所开发的纤维组织芯片在筛选抗纤维化方面的应用 化合物。在这个项目完成后，我们希望我们能开发出一种新的方法，大大加快抗纤维化疗法从实验室到临床的转化。我们相信，这种技术进步将对抗击纤维化疾病的实践产生积极影响。

 DESCRIPTION: Fibrosis, as seen in end-stage idiopathic pulmonary fibrosis (IPF), heart failure, liver cirrhosis and kidney disease, leads to organ failure and currently has no cure. Candidates for anti-fibrotic therapies have been identified; however, the translation of these laboratory discoveries to clinical practice is hindered by the slow disease progression and the high cost associated with the clinical trials. To justify the development of new therapies in prolonged and expensive clinical trials, an in-vitro screening platform that can provide early evidence of efficacy of the anti-fibrotic therapies is desired. However, failure of the existing in vitro models to fully recapitulate the physio-pathological characteristics of the fibrotic disease, such as the reduced Forced vital capacity (FVC) and major extracellular matrix (ECM) remodeling in lung fibrosis, has significantly delayed the development of much needed anti-fibrotic therapies. The objective of this project is to develop a microphysiological lung "fibrotic micro-tissue chip" device that can enable the early screening of anti-fibrotic therapies. We have recently adopted micro-fabrication techniques to assemble 3D fibroblast-populated submillimeter micro-tissues in arrays of poly (dimethylsiloxane) (PDMS) micro-wells. In each micro-well, fibroblasts spontaneously contract and assemble the matrix proteins, such as collagen, into aligned micro-tissues that anchor between a pair of cantilevers. Our recent preliminary data have shown that distinct epithelium and ECM layers can be formed in micro-tissues due to boundary condition guided self-assembly of the cells and ECMs. With appropriate biochemical and mechanical stimuli, fibrotic differentiation of the resident cells can be further induced in these micro-tissues. In this project, we will fully characterize the fibrotic propertiesof the induced micro-tissue, such as its compliance under simulated forced ventilation and the deposition of ECM. We will then test its utility against two drugs that are very recently approved by FDA to treat IPF. The Specific Aims of this application are: 1) Fabricate arrays of micro-tissues and induce fibrotic transition; 2) Calibrate the mechanical and histological characteristic of the fibrotic micro-tissues and optimize the system using a small set of training anti-fibrotic drugs; and 3) Evaluate the utility of the developed fibrotic tissue-chip in screening anti-fibrotic compounds. Upon completion of this project, it is our expectation that we will have developed a new approach that can significantly expedite the translation of anti-fibrotic therapies from the laboratories to the clinics. We are confident that such advancement in the technology will positively impact the practices to combat fibrotic diseases.