I-Corps: Human toxicity assay using synthetic embryo-like structures

I-Corps项目的更广泛的影响/商业潜力是开发一种可控和可扩展的微流体技术，在这种技术中，可以从人类干细胞中提取合成的人类胚胎样结构，用于定量毒性测试。所提出的技术的发展解决了对一个具有成本效益和高度预测性的测定平台的需求，以确定胚胎毒理学效应。目前的试验是用大鼠或小鼠等动物模型进行的。这些动物实验成本高昂，存在伦理问题，预测能力有限。这项拟议中的技术提供了与人类相关的“有机体水平”毒性测试。该方法允许定量测量和扰动，并与实时成像和高通量屏幕兼容。拟议的技术可以使用疾病和患者特异性干细胞进行疾病建模，也可以允许使用来自不同年龄和种族/民族群体的干细胞进行筛选，从而模拟对不同人群的药理学和化学化合物的反应。I-Corps项目的基础是开发一种可控和可扩展的微流控技术，其中来自人类干细胞的合成胚胎样结构可用于胚胎毒性测试。所提出的微流控装置允许在微流控装置内的规定位置精确定位人类干细胞集落。此外，该装置允许可溶性因子的精确时间递送，以驱动这些人类干细胞集落发育并组织成多细胞结构，其分子和细胞特征与早期植入后的人类胚胎具有显著的相似性。所提出的技术已被证明是可控的、可重复的、可扩展的，并且与实时成像和免疫细胞化学兼容，这可以很容易地应用于现有的高通量毒性筛选管道。该技术旨在提供基于比色/荧光试剂的药理学化合物对胚胎发育影响的高含量表型和形态学分析。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this I-Corps project is the development of a controllable and scalable microfluidic technology in which synthetic human embryo-like structures may be derived from human stem cells for quantitative toxicity testing. The development of the proposed technology addresses the need for a cost-effective and highly predictive assay platform to determine embryo toxicology effects. The current assay is conducted using animal models such as rats or mice. These animal tests are costly and have ethical issues and limited predictive powers. The proposed technology offers human-relevant, “organism-level” toxicity testing. The method allows quantitative measurements and perturbations and is compatible with live imaging and high-throughput screens. The proposed technology may use disease- and patient-specific stem cells for disease modeling, and also may allow screening using stem cells derived from different age and racial/ethnic groups, thus modeling responses to pharmacological and chemical compounds of diverse populations.This I-Corps project is based on the development of a controllable and scalable microfluidic technology in which synthetic embryo-like structures derived from human stem cells may be utilized for embryo toxicity testing. The proposed microfluidic device allows precise positioning of human stem cell colonies in prescribed locations within the microfluidic device. In addition, the proposed device allows precise temporal delivery of soluble factors to drive these human stem cell colonies to develop and organize into multicellular structures whose molecular and cellular features bear significant similarities to early post-implantation human embryos. The proposed technology has been shown to be controllable, reproducible, scalable, and compatible with live imaging and immunocytochemistry, which may allow easy adoption into the existing high-throughput toxicity screening pipelines. The technology is designed to provide high-content phenotypic and morphological profiling of the effects of pharmacological compounds on embryo development based on colorimetric/fluorescent reagents.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.