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

基本信息

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
项目类别：
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

项目摘要

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.

补充项目概要胎盘是母亲和胎儿之间的界面，因此控制着营养物质、废物的交换产品和治疗剂。药物对人体的潜在毒性、致畸性和治疗作用胎儿受母体器官（特别是肝脏和肾脏）的代谢和清除控制，它们穿过胎盘的运输以及胎盘的新陈代谢。鉴于这些功能的差异物种间的特性，动物模型测试并不总能预测药物对人类的不良影响怀孕。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，例如瑞德西韦（remdesivir），最近被研究发现对 COVID-19 有效。胎盘的合体滋养层主要负责药物转运和代谢。在我们的模型中，人类滋养层干细胞 (hTSC) 被接种在构建体的母体一侧，并允许合体化。对于这个补充项目，我们将介绍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室，并且检查在滋养层细胞中积累或转移到下室的产物，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 hTSC 和缺乏主动移动转运蛋白的突变型 hTSC 之间的功能差异药物进出滋养层细胞。最近的出版物报道了瑞德西韦的临床益处 COVID-19，人们对这种药物在怀孕期间的安全性非常感兴趣。瑞德西韦是一种核苷阻断 SARS-CoV-2 复制的类似物，可能是通过抑制依赖 RNA 的 RNA 合成，就像它对相关的中东呼吸综合征冠状病毒。瑞德西韦对怀孕的影响尚不清楚。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 并评估这些的影响瑞德西韦和恩曲他滨在 3D 胎盘中从母体转运至胎儿室的突变这些药物在滋养层细胞中的模型和积累。因此，拟议的研究将加速了解抗病毒药物穿过母胎屏障的胎盘渗透性并阐明转运蛋白参与开发安全且用于妊娠的治疗方法对母亲和胎儿均有效，符合父母补助金的目标。