Engineered tissue-based, high-throughput compound profiling

描述（由申请人提供）：心脏病经常导致心脏纤维化。在几乎所有的慢性心脏病病例中，心肌表现出由活化的心脏成纤维细胞发展的纤维化。这些成纤维细胞在健康的心脏中是静止的。由于成纤维细胞的细胞外基质沉积引起的间质纤维化增加了组织的硬度并损害了心脏舒张。发现可以逆转纤维化的药物治疗是一个关键的未满足的需求;目前没有这样的药物存在。该项目将开发一种新的高通量筛选平台，用于药物发现，测量活的工程组织样品及其对照的生理特性，在96孔板（PalpatorTM系统）中培养。该平台将测量药物诱导的工程组织生理特性的变化。第一阶段项目的重点是完成PalpatorTM筛选系统的开发，并从学术和工业合作者那里获得反馈。此外，将修改用于数据分析软件的算法，以降低故障率。将对修改后的软件进行beta测试，以确定其获得生理参数的有意义值的能力，这些生理参数用于指示用一组化合物处理已改变了工程组织的性质。与项目有关的软件的最后包装将外包，以便启动。第二阶段项目的重点是扩大基于组织的工程筛选系统，使其适合工业中的高通量应用。将与Engineering Industries，Inc.合作生产用于在96孔板中培养工程组织的组织培养耗材。（Verona，WI）。大规模的工程化组织生产将显著提高筛选效率。这种高效的Palpator筛选系统将用于分析50种常用心血管药物对工程心脏组织的影响。尽管工程心脏组织被构建为模拟天然心肌的生理特性，但分析已知的心血管药物将验证在药物发现和毒性化合物测试中采用工程心脏组织的效用。为了进一步验证工程组织模型，将使用Palpator系统筛选具有已知心血管效应的化合物库。一种新的表型筛选方案，其采用与高度收缩性成纤维细胞一起培养的工程化组织来模拟纤维化心脏和“正常”工程化心脏组织，将用于鉴定降低纤维化工程化组织的收缩性但仍保持正常工程化心脏组织的健康收缩活性的化合物。工程组织模型和Palpator筛选设备的结合将加速药物发现，并减少广泛的动物研究的需求（和相关成本）。公共卫生相关性：在今天的美国，大约有500万人患有心脏病，这是最普遍的慢性病之一，也是心脏病发作的头号并发症。该提案描述了一种全新的方法来测试潜在的药物，通过测量药物是否改变心脏组织模型的行为来了解它们是否可以帮助修复心脏或是否对身体有毒。这项研究可能在加速药物发现和降低成本方面提供突破，因为它快速，高通量，并可能减少对动物试验的需求。

DESCRIPTION (provided by applicant): Heart disease frequently leads to cardiac fibrosis. In almost all cases of chronic heart disease, the myocardium exhibits fibrosis developed by activated cardiac fibroblasts. These fibroblasts are quiescent in the healthy heart. Interstitial fibrosis due to extracellular matrix deposition by fibroblasts increases the stiffness of the tissue and impairs cardiac relaxation. Discovering pharmaceutical treatments that can reverse fibrosis is a critical unmet need; no such drugs currently exist. This project will develop a novel high-throughput screening platform for drug discovery that measures the physiological properties of live, engineered tissue samples, and their controls, cultured in 96-well plates(the PalpatorTM system). This platform will measure drug-induced changes in the physiological properties of engineered tissues. The Phase I project focuses on completing the development of the PalpatorTM screening system and obtaining feedback from academic and industrial collaborators. In addition, the algorithm used for the data analysis software will be modified to reduce failure rates. The modified software will be beta tested for its ability to obtain meaningful values for the physiological parameters used to indicate that treatment with the panel of chemical compounds has altered the properties of the engineered tissues. The final packaging of the project-related software will be outsourced for its launch. The Phase II project focuses on scaling up the engineered tissue-based screening system to make it amenable to high-throughput applications in industry. The tissue culture consumables for growing engineered tissues in 96-well plates will be produced in collaboration with Engineering Industries, Inc. (Verona, WI). The scaled-up engineered tissue production will significantly improve screening efficiency. This highly efficient Palpator screening system will be used to profile the effects of 50 commonly prescribed cardiovascular drugs on engineered heart tissues. Although the engineered heart tissues are constructed to mimic the physiological properties of native heart muscles, profiling known cardiovascular drugs will validate the utility of employing engineered heart tissues in drug discovery and toxic compound testing. To further validate the engineered tissue model, a library of compounds with known cardiovascular effects will be screened using the Palpator system. A novel phenotypic screening protocol that employs both engineered tissues cultured with highly contractile fibroblasts to mimic the fibrotic heart and 'normal' engineered heart tissues will be used to identify chemical compounds that reduce the contractility of the fibrotic engineered tissues but yet maintain the healthy contractile activities of normal engineered heart tissues. The combination of the engineered tissue models and the Palpator screening device will accelerate drug discovery and reduce the need (and associated costs) of extensive animal studies. PUBLIC HEALTH RELEVANCE: In the United States today, about 5 million people suffer from heart disease, one of the most prevalent chronic conditions and the number one complication of heart attacks. This proposal describes an entirely new method for testing potential drugs to learn if they can help repair the heart or if they are toxic to the body, by measuring whether a drug changes how a heart tissue model behaves. This research may provide a breakthrough in accelerating drug discovery and reducing costs, because it is rapid, high-throughput, and may reduce the need for animal testing.