Modeling human trophoblast-NK cell interactions in term and preterm birth

项目摘要/摘要 人类胎盘是一种半同种异体组织，其生长和发育需要耐受 母体免疫系统。事实上，孕妇的免疫系统在怀孕期间面临着一个挑战： 保持对外来胎儿同种异体抗原的耐受性，同时对潜在的 母胎交界处的病原体。胎盘细胞逃避母体的机制 人们对免疫识别知之甚少，尤其是在人类怀孕的背景下。子宫 衬里，称为蜕膜，是一个特别未被研究和重要的微环境，因为它是 胎盘细胞称为绒毛外滋养细胞(EVT)与母体密切接触的界面 免疫细胞，其中蜕膜自然杀伤(DNK)细胞最为丰富。EVT是高度侵入性的 母体子宫内壁正确重塑所需的细胞，包括血管 导致母体血液流向胎盘的重塑。之间的相互作用 已知胎盘EVT和蜕膜白细胞通过EVT和 限制宫腔静脉血栓侵犯子宫壁的范围。事实上，早产的问题可能是由 DNK细胞的不适当反应。不幸的是，DNK和滋养层细胞之间的相互作用 在怀孕期间很难学习，因为无法进入蜕膜室，在那里 重要的互动发生了。虽然动物模型已经为这些过程提供了一些见解，但他们 不要准确地模拟人类胎盘和怀孕。最初的R01提案的目标是 评估足月和早产的蜕膜细胞数量，然后将这些知识与 再生医学最新技术发展DNK-EVT体外模型的研究 互动。具体地说，我们建议产生匹配的母体和胎盘诱导的多能性 干细胞(IPSC)，并将这些细胞分别分化为DNK细胞和EVT细胞，以便建立 这两种细胞类型之间的相互作用。鉴于已知的男性胎儿性别与 早产时，我们把精力集中在鉴定、储存和重新编程细胞上。 妈妈：原始申请中的男婴配对。然而，要充分了解 对于sPTB的易感性，最好将这些数据与母亲：女婴配对的数据进行比较。 因此，我们现在建议对来自母亲：女性胎盘的细胞进行表征、储存和重新编程。 此行政补充资料的配对。成功完成这项提案将建立一个 研究母体免疫相互作用的可重复性和可操作性的模型系统 系统和男性和女性胎盘。

Project Summary/Abstract The human placenta is a semi-allogeneic tissue whose growth and development requires tolerance by the maternal immune system. In fact, the maternal immune system faces a challenge during pregnancy: to maintain tolerance toward foreign fetal alloantigens while simultaneously staging a response to potential pathogens at the maternal-fetal interface. The mechanisms through which placental cells evade maternal immune recognition are poorly understood, particularly in the context of human pregnancy. The uterine lining, called decidua, is a particularly-understudied and important microenvironment, because it is the interface where placental cells called extravillous trophoblast (EVT) come in close contact with maternal immune cells, of which decidual natural killer (dNK) cells are the most abundant. EVT are highly invasive cells which are required for proper remodeling of the maternal uterine lining, including vascular remodeling which leads to establishment of maternal blood flow to the placenta. Interactions between placental EVT and decidual leukocytes are known to facilitate maternal vascular remodeling by EVT and limit the extent of EVT invasion into the uterine wall. Indeed, problems in preterm birth could result from inappropriate responses by dNK cells. Unfortunately, interactions between dNK and trophoblasts are difficult to study in an ongoing pregnancy, due to lack of access to the decidual compartment, where these important interactions occur. While animal models have offered some insights into these processes, they do not accurately model human placentation and pregnancy. The goal of the original R01 proposal is to evaluate the decidual cell population in both term and preterm birth, then to combine this knowledge with the latest technologies in regenerative medicine to develop in vitro models for the study of dNK-EVT interactions. Specifically, we proposed to generate matched maternal and placental induced pluripotent stem cells (iPSC), and differentiate these cells into dNK cells and EVT, respectively, in order to model interactions between these two cell types. Given the known association between male fetal sex and risk of preterm birth, we had focused our efforts on characterizing, banking, and reprogramming cells from mom:male baby pairs in the original application. However, to fully understand mechanisms that predispose to sPTB, it would be best to compare these data to those from mom:female baby pairs. Therefore, we now propose to characterize, bank, and reprogram cells from mom:female placenta pairs for this administrative supplement. Successful completion of this proposal will establish a reproducible and manipulatable model system for studying interactions between the maternal immune system and both the male and female placenta.