Stem Cell-based Human Placenta-on-a-Chip Using 3D Bioprinting

使用 3D 生物打印技术开发基于干细胞的人类胎盘芯片

• 批准号: 10177137
• 负责人: SHAOCHEN CHEN
• 金额: $ 22.27万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-30 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10177137
• 关键词: 2019-nCoV; 3-Dimensional; ABCC1 gene; Active Biological Transport; Adverse drug effect; Animal Model; Antiviral Agents; Blood; Blood capillaries; COVID-19; CRISPR/Cas technology; Cell Line; Cell Survival; Cells; Clinical; Coculture Techniques; Drug Transport; Endothelial Cells; Endothelium; Fetus; Fibroblasts; Gelatin; Human; Kidney; Knock-out; Liver; Metabolism; Methacrylates; Middle East Respiratory Syndrome Coronavirus; Modeling; Mothers; Mutate; Mutation; Nutrient; Organ; Perfusion; Permeability; Pharmaceutical Preparations; Physiological; Placenta; Pregnancy; Property; Publications; RNA; RNA chemical synthesis; Reporting; Research; Role; Side; Stromal Cells; Syncytiotrophoblast; System; Teratogenic effects; Testing; Therapeutic Agents; Therapeutic Effect; Therapeutic Uses; Time; Toxic effect; Villous; Waste Products; base; bioprinting; cell type; drug metabolism; emtricitabine; experimental study; fetal; interest; medication safety; monolayer; mutant; nucleoside analog; parent grant; parent project; remdesivir; stem cells; therapeutic development; trophoblast

Supplement Project Summary The placenta is the interface between mother and fetus, and as such controls the exchange of nutrients, waste products, and therapeutic agents. The potential toxicity, teratogenicity, and therapeutic effects of drugs on the fetus are controlled by their metabolism and clearance by maternal organs (particularly the liver and kidney), their transport across the placenta, and their metabolism by the placenta. Given functional differences in these properties between species, testing in animal models does not always predict adverse drug effects in human pregnancy. Here, we propose to extend our parent project by adapting our model to investigate the role of the placenta in drug transport, specifically, transport of nucleoside analogs such as remdesivir, which was recently found to have efficacy against COVID-19. The syncytiotrophoblast layer of the placenta is largely responsible for drug transport and metabolism. In our model, human trophoblast stem cells (hTSCs) are seeded on the maternal side of the construct and allowed to syncytialize. For this supplement project, we will introduce drugs that are known or suspected to be actively transported across the placenta into the upper chamber, which represents the maternal blood space, and examine the products that accumulate in the trophoblast cells or are transferred to the lower chamber, which represents the placental villous capillaries on the fetal side. We propose to use this model to evaluate the functional differences between wild type hTSCs and mutant hTSCs deficient in transporters that actively move drugs in and out of trophoblast cells. With the recent publication reporting clinical benefit from remdesivir for COVID-19, there is intense interest in the safety of this medication in pregnancy. Remdesivir is a nucleoside analog that blocks SARS-CoV-2 replication, likely by inhibiting RNA-dependent RNA synthesis, as it does for the related MERS coronavirus. The effects of remdesivir in pregnancy are unknown. In this project, we aim to use CRISPR-Cas9 to knock out ENT1, CNT1, OCTN1, and MRP1 in hTSCs and evaluate the effects of these mutations on transport of remdesivir and emtricitabine from the maternal to fetal compartment in our 3D placenta model and accumulation of these drugs in the trophoblast cells. The proposed research will therefore accelerate the understanding of transplacental permeability of antiviral drugs across the maternal-fetal barrier and elucidate the involvement of transporters towards the development of therapeutics for use in pregnancy that are safe and effective for both the mother and fetus, consistent with the objectives of the parent grant.

补充项目摘要 胎盘是母亲和胎儿之间的界面，因此控制着营养物质、排泄物的交换 产品和治疗剂。药物的潜在毒性、致畸性和治疗作用 胎儿受母体器官(特别是肝脏和肾脏)的新陈代谢和排泄控制， 它们通过胎盘运输，通过胎盘代谢。考虑到这些方面的功能差异 不同物种之间的特性，在动物模型中的测试并不总是预测人类的药物不良反应 怀孕了。在这里，我们建议通过调整我们的模型来扩展我们的父项目，以调查 胎盘在药物转运中的作用，特别是核苷类似物的转运，如最近发现的瑞希韦 被发现对新冠肺炎有疗效。 胎盘的合体滋养层主要负责药物的运输和代谢。在我们的 模型中，人类滋养层干细胞(HTSCs)被种植在构建物的母体一侧，并允许 合胞化。对于这一补充项目，我们将推出已知或怀疑活跃的药物 穿过胎盘进入代表母体血液空间的上腔，以及 检查在滋养层细胞中积累或转移到下腔的产物， 代表胎儿一侧的胎盘绒毛毛细血管。我们建议使用该模型来评估 野生型hTSCs与突变型hTSCs在主动运动转运体缺陷方面的功能差异 药物进出滋养层细胞。最近的一篇文章报道了瑞希韦的临床益处 新冠肺炎，人们对这种药物在怀孕期间的安全性非常感兴趣。雷米地韦是一种核苷 类似物，可能通过抑制依赖RNA的RNA合成来阻止SARS-CoV-2的复制，就像它对 相关的MERS冠状病毒。雷米西韦在怀孕期间的作用尚不清楚。在这个项目中，我们的目标是使用 CRISPR-CAS9敲除hTSC中的ENT1、CNT1、OCTN1和MRP1并评估这些基因的效果 瑞希韦和恩曲他滨在3D胎盘中从母体转运到胎儿体内的突变 这些药物在滋养层细胞中的模型和蓄积。因此，拟议的研究将加快 抗病毒药物经胎盘对母胎屏障通透性的认识与阐明 转运蛋白参与开发用于妊娠的安全和 对母亲和胎儿都有效，与父母赠款的目标一致。