Integration of a kidney module into a 4-organ human-on-a-chip system.

项目摘要 我们对Hesperos的总体战略是利用微生理系统与功能 读数以建立能够对毒性化学品和候选药物进行复杂分析的平台 和临床前试验期间的有效性，最初的重点是预测性毒性。这是一项基于 该公司正在开发低成本的体外系统，该系统利用一种新的“无泵”微生理平台 在2014年6月10日的美国专利8,748,180B2中描述。每个隔间代表一个组织或器官 通过一种模拟血液的无血清介质连接起来。无泵集成系统，使用 摇摆运动泵送无血清细胞培养液，降低了流体回路的复杂性和成本 设计并简化了设备的设置和操作。希克曼开发了微电极阵列和 集成在芯片上的悬臂系统，允许非侵入性的电子和机械读数。 我们将利用该系统的微流控分析组件进行快速和灵敏的生物标记物(化学， 蛋白质和小分子)评估。然而，为细胞健康而监测的生物标志物的数量 而在我们的系统中，由于使用了功能读出，功能将大大减少。我们已经建造了 代表心脏、肌肉和肝脏功能的生理系统，在Hesperos和 希克曼的实验室被制药和化妆品公司。我们已经证明了多器官毒性在 由神经、心脏、肝脏和肌肉组成的四器官系统。该系统是世界上第一个 微流控模型展示促进4种细胞存活和功能能力的能力 在延长的培养时间内，在无血清介质中相互连接的组织模块。更多的证据表明 我们平台的商业可行性在于，我们刚刚从National获得了SBIR第二阶段拨款 先进翻译科学中心(NCATS)将先进的制造技术应用于我们的 与国家标准与技术研究所(NIST)合作的基本四器官平台。 在这项提案中，我们扩展了4个器官系统，以包括人类肾脏的模型，作为迈向 更完整的ADMETox模型。我们的第一个目标将是设计、制造和测试一个肾脏模块来建模 通过肾脏排出的几个方面。然后，我们将把肾脏模块与现有的4个器官合并 模块。这需要增加另一层，以允许透析流模拟排泄通过 肾脏。肾脏将有肾小球和近端小管隔间。用于直接测量的电极 TEER(跨皮细胞电阻)将被整合到肾脏模块中。现有的系统将 测量骨骼肌和心肌力的功能读数，以及神经元和 有图案的心肌细胞。该系统将通过已被证明有效的药物和化学品进行验证 肾功能。

Project Summary Our overall strategy for Hesperos is to utilize microphysiological systems in combination with functional readouts to establish platforms capable of sophisticated analysis of chemicals and drug candidates for toxicity and efficacy during pre-clinical testing, with initial emphasis on predictive toxicity. This is a service based company and is developing low-cost in vitro systems utilizing a novel “pumpless” microphysiological platform described in US Patent 8,748,180B2, June 10, 2014. The compartments each representing a tissue or organ are connected by a serum-free medium which mimics the blood. The pumpless integrated system, using a rocking motion to pump the serum-free cellular medium, reduces the complexity and cost of the fluidic circuit design and simplifies set-up and operation of the device. Hickman has developed microelectrode arrays and cantilever systems that are integrated on chip that allows for noninvasive electronic and mechanical readouts. We will utilize microfluidic analytical components with this system for rapid and sensitive biomarker (chemical, protein and small molecule) assessment. However, the number of biomarkers to be monitored for cell health and function will be greatly reduced in our systems from use of the functional readouts. We have constructed physiological systems that represent cardiac, muscle and liver function that are being used at Hesperos and in Hickman’s lab by pharmaceutical and cosmetic companies. We have demonstrated multi-organ toxicity in the 4-organ system composed of neuronal, cardiac, liver and muscle components. This system is the first microfluidic model to demonstrate the capacity to promote the survival and functional competence of 4 interconnected tissue modules in serum-free medium over extended culture periods. Further evidence of the commercial viability of our platform is that we have just been awarded a Phase II SBIR grant from National Center for Advancing Translational Sciences (NCATS) to apply advanced manufacturing techniques to our basic 4-organ platform in collaboration with National Institute of Standards and Technology (NIST). In this proposal we extend the 4 organ system to include a model of the human kidney as a step toward a more complete ADMETox model. Our first aim will be to design, fabricate and test a kidney module to model aspects of elimination through the kidney. We will then incorporate the kidney module with the existing 4 organ module. This requires the addition of another layer to allow flow of a dialysis stream to mimic excretion through the kidney. The kidney will have glomerulus and proximal tubule compartments. Electrodes to directly measure TEER (transepithelial electrical resistance) will be integrated into the kidney module. The existing system will measure functional readouts of force for skeletal muscle and cardiac, and electrical activity of neurons and patterned cardiomyocytes. The system will be validated with drugs and chemicals that have shown to effect kidney function.