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、废物和蛋白质的交换。 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）的代谢和清除控制， 它们通过胎盘的运输和胎盘的代谢。鉴于这些功能的差异， 动物模型中的测试并不总是预测人类中的药物不良反应 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，如最近被 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 在该模型中，将人滋养层干细胞（hTSC）接种在构建体的母体侧，并允许其 合胞化对于这一补充项目，我们将引入已知或疑似积极的药物， 穿过胎盘进入代表母体血液空间的上腔室， 检查滋养层细胞中积累的产物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hTSCs和缺乏主动移动的转运蛋白的突变型hTSCs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Remdesivir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。Remdesivir是一种核苷 类似物，阻断SARS-CoV-2复制，可能是通过抑制RNA依赖的RNA合成，就像它对 与MERS冠状病毒有关。Remdesivir对妊娠的影响尚不清楚。在这个项目中，我们的目标是使用 CRISPR-Cas9敲除hTSC中的ENT 1、CNT 1、OCTN 1和MRP 1，并评估这些基因的作用 在我们的3D胎盘中，Remdesivir和恩曲他滨从母体转运至胎儿室的突变 模型和这些药物在滋养层细胞中的积累。因此，拟议的研究将加速 了解抗病毒药物穿过母胎屏障的经胎盘渗透性，并阐明 转运蛋白参与开发用于妊娠的安全且 对母亲和胎儿都有效，符合父母补助金的目标。