Probing and modeling decidual natural killer cells in preterm birth

早产中蜕膜自然杀伤细胞的探测和建模

• 批准号: 10607933
• 负责人: Virginia Chu Cheung
• 金额: $ 6.97万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-11-01 至 2024-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10607933
• 关键词: Alloantigen; Allogenic; Animal Model; Apoptosis; Basal Plate; Biological Assay; Biological Models; Blood; Blood flow; CXCL12 gene; Catalogs; Cell Communication; Cell physiology; Cells; Chemicals; Coculture Techniques; Data; Decidua; Decidual Cell; Decidual Cell Reactions; Development; Disease; Endometrial; Endometrium; Environment; Equilibrium; Fetus; Fibroblasts; First Pregnancy Trimester; Flow Cytometry; Galactose Binding Lectin; Gene Expression; Gestational Age; Growth and Development function; Heterogeneity; Human; Hypoxia; IL8 gene; Immune; Immune system; Individual; Inflammatory; Ionomycin; K562 Cells; Labor Onset; Leukocytes; Location; Maintenance; Maternal-Fetal Exchange; Measures; Membrane; Methods; Modeling; Natural Killer Cells; Nature; Patients; Phenotype; Placenta; Placentation; Play; Population; Pregnancy; Pregnancy Outcome; Premature Birth; Premature Labor; Process; Protocols documentation; Publishing; Resources; Role; Series; Staging; Surface; Term Birth; Time; Tissues; Uterus; Vascular Endothelial Growth Factors; Vascular remodeling; Work; base; cytokine; cytotoxicity; differentiation protocol; fetal; gene function; healthy pregnancy; human model; in vitro Model; induced pluripotent stem cell; insight; pathogen; peripheral blood; response; single-cell RNA sequencing; stem cell differentiation; transcriptome; transcriptome sequencing; transcriptomics; trophoblast

Project Summary/Abstract The human placenta is a semi-allogeneic tissue whose growth and development requires tolerance by the maternal immune system. How this relationship is compromised in the setting of spontaneous preterm birth (sPTB) is currently unknown. In a normal pregnancy, placental cells come in close contact with maternal blood and uterine tissues yet are able to evade immune recognition throughout gestation. During this time, the maternal immune system maintains a balance of tolerance toward foreign fetal alloantigens while simultaneously staging a response to potential pathogens at the maternal-fetal interface. The nature of such maternal-fetal cellular interactions is poorly understood, particularly when it goes awry in the setting of spontaneous preterm birth (sPTB). The uterine lining, called decidua, is a particularly understudied microenvironment, containing many maternal immune cells, of which decidual natural killer (dNK) cells are the most abundant. It is at this interface where placental cells called extravillous trophoblast (EVT) come in close contact with maternal immune cells. Invasion of the decidua by fetal EVT is 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, such as dNK, are known to facilitate maternal vascular remodeling by EVT and limit the extent of EVT invasion into the uterine wall. Indeed, problems in sPTB could result from inappropriate responses by dNK cells. Unfortunately, dNK function, including dNK-trophoblast crosstalk, is difficult to study in an ongoing pregnancy, due to lack of access to the decidual compartment, where these important interactions occur. Additionally, very few studies have characterized dNK at term when the pregnancy outcome is known. While animal models have offered some insights into these processes, they do not accurately model human placentation and pregnancy. This proposal aims to functionally and transcriptomically characterize dNK subpopulation(s) in term, with or without labor, and preterm placenta, then use induced pluripotent stem cells (iPSC) to model dNK, in the setting of both term and preterm birth. Our team has established optimized methods for differentiation of iPSC into both peripheral blood NK cells and EVT. We will now optimize a protocol for iPSC differentiation into decidual NK cells, comparing them directly to primary cells. Successful completion of this proposal will reveal underlying mechanisms through which abnormal dNK activation contributes to spontaneous preterm birth, and establish a renewable and manipulatable model of dNK at the human maternal-fetal interface.

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。在自然早产的情况下，这种关系是如何受到损害的 (SPTB)目前尚不清楚。在正常妊娠中，胎盘细胞与母体血液密切接触 而子宫组织在整个怀孕过程中仍然能够逃避免疫识别。在这段时间里，母亲 免疫系统在分期的同时维持对外来胎儿同种异体抗原的耐受平衡 在母胎界面对潜在病原体的反应。这种母胎细胞的性质 人们对相互作用知之甚少，特别是在自发早产的环境中出现问题时 (SPTB)。子宫内膜，称为蜕膜，是一种特别未被研究的微环境，包含许多 母体免疫细胞，其中蜕膜自然杀伤(DNK)细胞最为丰富。就在这个界面上 胎盘细胞称为绒毛外滋养细胞(EVT)，与母体免疫细胞密切接触。 胎儿EVT侵入蜕膜是母体子宫内壁正确重塑所必需的，包括 血管重塑，导致母体血液流向胎盘。之间的相互作用 众所周知，胎盘EVT和蜕膜白细胞，如DNK，通过以下方式促进母体血管重构 EVT和限制EVT侵犯子宫壁的范围。事实上，sptb中的问题可能源于 DNK细胞的不适当反应。不幸的是，包括DNK-滋养细胞串扰在内的DNK功能是 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。此外，很少有研究将DNK描述为怀孕时的足月 结果是已知的。虽然动物模型已经为这些过程提供了一些见解，但它们并不准确 人体胎盘和妊娠模型。这项提案旨在从功能和转录的角度描述 DNK亚群(S)在足月、有无分娩、早产胎盘，然后使用诱导多能干细胞 细胞(IPSC)来模拟DNK，在足月和早产的情况下。我们的团队已经建立了优化的 IPSC向外周血NK细胞和EVT细胞分化的方法。我们现在将优化一个协议 IPSC分化为蜕膜NK细胞，直接与原代细胞比较。成功完成 这一提议将揭示DNK异常激活对 自然早产，并在人类建立可再生和可操作的DNK模型 母胎界面。