Establishing an In Vitro Embryotoxicity Risk Classification System Based on Human Cardiac Organoid Model

项目摘要 目前，药物胚胎毒性对安全妊娠的风险尚未完全确定，因此许多孕妇 暴露在未知对胎儿发育影响的药物中。许多药物仍然没有得到很好的理解 关于它们对人类器官发生的影响，也没有一种公认的人类胚胎毒性药物 提供筛查平台。目前，人类诱导多能干细胞(HiPSCs)已被提出用于 人体相关药物毒性筛选。然而，在2D上使用HiPSC进行维护和分化 培养法不是一种理想的胚胎毒性分析方法，因为它们无法预测药物对3D组织的毒性 形态发生，这可能导致产前晚期表现出的结构畸形 发展。随着干细胞类器官概念的出现，这些发育中组织的3D培养意味着 与不同机构建立其特色组织的方式相似 发展。因此，本方案的总体目标是建立一种基于HiPSC的体外心脏器官 基于药物对HiPSC生长、心脏分化和早期胚胎毒性测试的模型 心脏的形成，所以我们可以建立一个风险分类系统，以便更准确地评估人类的特异性 药物对早期胚胎发育的影响。为实现这一目标，我们将追求三个具体目标。在目标1中， 我们将通过研究生物物理限制对心脏器官模型的影响来优化心脏器官模型。 心脏类器官的形成和功能。在目标2中，我们将验证心脏有机化合物的胚胎毒性。 与现行标准斑马鱼全胚胎培养法进行比较。通过测试一个“培训” 一组已知胚胎毒性水平的化学物质，我们将更好地校准人类心脏的药物反应 基于各种终点的有机化合物评价参数。在目标3中，我们将建立一个新的生物统计学 基于线性判别分析的胚胎毒性风险分类预测模型。我们预见到 这种体外心脏器官模型可以改进传统药物的筛选 在怀孕期间使用。

Project Summary Currently, drug embryotoxicity risk for safe pregnancy is not well established, thus many pregnant women are exposed to the drugs with unknown effects on fetus development. Many drugs are still neither well understood regarding their effects on human organogenesis, nor is there a well-established human embryotoxicity drug screening platform available. Currently, human induced pluripotent stem cells (hiPSCs) have been proposed for human-relevant drug toxicity screening. However, the use of hiPSC maintenance and differentiation on 2D culture is not an ideal embryotoxicity assay due to their inability to predict the drug toxicity on 3D tissue morphogenesis, which potentially leads to the structural malformations manifested in late prenatal fetus development. With the emergent concept of stem cell organoids, these 3D cultures of developing tissues imply the similarity to the manner in which different organs establish their characteristic organization during development. Therefore, the overall goal of this proposal is to establish an in vitro hiPSC-based cardiac organoid model for embryotoxicity testing based on the drug effects on hiPSC growth, cardiac differentiation, and early heart formation, so we can establish a risk classification system for more precise assessment of human-specific drug effects on early embryonic development. To achieve this goal, we will pursue three specific aims. In Aim 1, we will optimize the cardiac organoid model by investigating the effects of biophysical confinement on the formation and function of cardiac organoids. In Aim 2, we will validate the cardiac organoid-based embryotoxicity assay by comparing to well-established standard zebrafish whole embryo culture assay. By testing a “training set” of chemicals with known embryotoxicity level, we will better calibrate the drug response from human cardiac organoids based on a variety of endpoint evaluation parameters. In Aim 3, we will establish a new biostatistical predictive model based on linear discriminant analysis for embryotoxicity risk classification. We envisage that this in vitro cardiac organoid model can improve traditional pharmaceutical screening for the drugs that will be administered during pregnancy.