Bioprinted Human Ventricles for In Vitro Modeling of Cardiac Arrhythmias

在过去的40年里，近45%的药物退出市场是由于心脏安全性 这些问题导致生物制药研发过程的成本不断增加，生产率不断下降。 虽然药物诱导的心脏毒性的机制因药物和靶点而异，但最常见和最常见的是心脏毒性。 危险表现为心律失常和心源性猝死。生物制药行业已经严重 投资于对心脏毒性作用敏感的新工具，然而，目前的临床前模型是一种 在结构、组成和功能复杂性方面的妥协， 预测人类心脏电生理学。此外，了解患者特定的风险因素，包括 遗传易感性、年龄、性别和基础心血管疾病（例如纤维化、缺血、梗死） 有助于药物诱导的促孕激素状态需要开发全新的体外模型 冲动传导障碍在这项计划中，我们的目标是开发一种新的生物工程人类心室 作为一个预测性的体外模型，用于确定药物诱导的促心脏病的风险。到 克服目前的局限性，FluidForm公司与卡内基梅隆大学合作，将开发一种新的 悬浮水凝胶的自由形式可逆包埋（FRESH）3D生物打印左心室模型， 重建了心室肌的层状结构，并具有定制的结构和组成， 促孕性疾病状态。我们的初步数据确定，我们可以建立一个功能性心室， 环周肌纤维排列，各向异性动作电位传播，明显的心律失常特征 包括转子和多个传播波，以及复杂的生物力学响应，包括壁 增厚在这里，我们将通过两项研究来改善心室性能，以用于生物制药研发过程 目标。首先，我们将建立FRESH 3D生物打印的人类心室模型对已知的心室模型的基线灵敏度。 产生促孕激素化合物并产生工业标准剂量-反应曲线。二是 通过控制心肌细胞和胶原结构模拟纤维化疾病， 并掺入具有已知传导突变的iPS衍生的人心肌细胞。这将使我们能够 实现患者特异性疾病模型，其显示左移的剂量-反应曲线， 促孕化合物第一阶段概念验证的成功将为第二阶段的成功奠定坚实的基础。 II SBIR项目，将在体外高内容物环境中验证完整的FRESH 3D打印心室模型 成像平台，以评估电生理学和生物反应，并提供急需的，行业- 在药物开发中准确预测人类心律失常的领先能力。

Over the past 40 years nearly 45% of drugs withdrawn from the market have been due to cardiac safety concerns, contributing to the ever increasing cost and declining productivity of the biopharma R&D process. While the mechanisms of drug-induced cardiotoxicity vary widely by drug and target, the most common and dangerous manifestation is cardiac arrhythmia and sudden cardiac death. The biopharma industry has heavily invested in new tools that are sensitive to cardiotoxic effects, however, current preclinical models are a compromise in the structural, compositional, and functional complexity necessary to recapitulate and be predictive of human cardiac electrophysiology. Further, understanding how patient-specific risk factors including genetic predisposition, age, sex, and underlying cardiovascular disease (e.g. fibrosis, ischemia, infarction) contribute to a drug-induced proarrhythmogenic state requires the development of entirely new in vitro models of impulse conduction disorders. In this proposal our objective is to develop a new bioengineered human ventricle as a predictive in vitro model for identifying drug-induced proarrhythmogenic risks in the human heart. To overcome current limitations, FluidForm, Inc in collaboration with Carnegie Mellon University will develop a new freeform reversible embedding of suspended hydrogels (FRESH) 3D bioprinted left ventricle model that recreates the laminar architecture of ventricular myocardium and has tailored structure and composition to mimic proarrhythmogenic disease states. Our preliminary data establishes that we can build a functional ventricle with circumferential myofiber alignment, anisotropic action potential propagation, distinct arrhythmia features including rotors and multiple propagating waves, and complex biomechanical responses including wall thickening. Here we will improve ventricle performance for use in the biopharma R&D process via two research aims. First, we will establish baseline sensitivity of the FRESH 3D bioprinted human ventricle model to known proarrhythmogenic compounds and generate industry-standard does-response curves. Second, we will demonstrate tunable sensitivity by controlling cardiomyocyte and collagen architecture to mimic fibrotic disease and incorporate iPS-derived human cardiomyocytes with known conduction mutations. This will allow us to achieve patient-specific disease models that show dose-response curves that are left-shifted for proarrhythmogenic compounds. Phase I proof-of-concept success will provide a strong foundation for a Phase II SBIR project that will validate the complete FRESH 3D printed ventricle model in an in vitro high-content imaging platform to assess electrophysiology and biological response, and provide a critically needed, industry- leading capability to accurately predict human arrhythmias in drug development.