Modeling Preeclampsia Using Human Pluripotent Stem Cells

使用人类多能干细胞模拟先兆子痫

• 批准号: 10394333
• 负责人: Mariko Horii
• 金额: $ 24.4万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10394333
• 关键词: ATAC-seq; Abnormal placentation; Affect; Alkaline Phosphatase; Biological Assay; Biological Models; Biological Specimen Banks; CDX2 gene; Cardiovascular system; Caring; Cell Line; Cells; Characteristics; Chorionic villi; Chromatin; Coculture Techniques; Complement; Complication; DNA Methylation; Decidua; Defect; Development; Disease; Disease model; Down Syndrome; Environmental Risk Factor; Epigenetic Process; Epithelial Cells; Etiology; Event; Fetal Growth; Fetal Growth Retardation; Functional disorder; Gene Expression; Gene Expression Profile; Generations; Gestational Age; Goals; Growth; Growth Factor; Histone H3; Human; Human Chorionic Gonadotropin; Hypertension; Hypoxia; Immune; In Vitro; Inflammatory; Injury; Interferon Type II; Interleukin-10; Lead; Leukocytes; Life; MMP2 gene; Measurement; Mesenchymal; Mesenchymal Stem Cells; Methods; Methylation; Modality; Modeling; Modification; Natural Killer Cells; Nature; Neonatal Mortality; Nervous system structure; Nutrient; Organ; Oxidative Stress; Oxygen; Pancreas; Pathogenesis; Perinatal; Phenotype; Placenta; Placental Insufficiency; Placentation; Pre-Eclampsia; Pregnancy; Pregnancy Complications; Premature Birth; Proliferation Marker; Proteinuria; Protocols documentation; Reproducibility; Sendai virus; Stains; Stem cell pluripotency; Study models; Syncytiotrophoblast; Technology; Teratoma; Testing; Therapeutic; Time; Tissues; Umbilical cord structure; Uterus; Vascular remodeling; Villous; Woman; Work; adverse outcome; base; bone morphogenetic protein 4; cytokine; cytotrophoblast; diagnostic tool; early onset; embryonic stem cell; epigenetic memory; epigenome; experimental study; gene function; genome-wide; human disease; human embryonic stem cell; human pluripotent stem cell; implantation; improved; in vivo; indexing; induced pluripotent stem cell; inflammatory milieu; macrophage; maternal morbidity; matrigel; method development; neonatal morbidity; novel diagnostics; pathophysiology of preeclampsia; perinatal period; pluripotency; pregnancy hypertension; stem cell model; stem cells; therapeutic target; transcriptome sequencing; trophoblast; trophoblast stem cell

Project Summary/Abstract Placenta is a transient organ that connects mom and baby during pregnancy, and supplies nutrients and oxygen for intrauterine fetal growth. It has been known that abnormal placentation can lead to preeclampsia (PE), a major complication affecting 2-8% of all pregnancies, presenting as new onset hypertension and proteinuria in the latter half of pregnancy. PE is associated with growth restriction and preterm delivery of the baby, both of which are associated with adverse outcomes, in both the perinatal period and later in life. Many studies point to abnormal differentiation of placental epithelial cells, called trophoblast, as the etiology of PE. Specifically, development of invasive extravillous trophoblast (EVT) and/or their function are affected in PE, leading to shallow implantation and placental insufficiency. However, while these are some accepted hypotheses about the nature of the disease, the underlying pathophysiology is still not fully understood mainly due to the lack of a truly representative model system. Over the past decade, the development of methods to generate induced pluripotent stem cells (iPSC) has led to the ability to model numerous human diseases, including those originating in the cardiovascular and nervous systems and the pancreas. In addition, over the same time period, multiple groups, including ours, have shown that iPSCs can be differentiated into trophoblast, using the growth factor, bone morphogenetic protein 4 (BMP4). My own work has in fact provided proof-of-concept that defects in trophoblast differentiation, specifically those associated with Trisomy 21, can be modeled using iPSCs. I therefore plan to apply this technology to model placental dysfunction in PE, with the following three aims: 1) I will establish iPSCs by reprogramming umbilical cord mesenchymal stem cells (UC-MSCs) from six PE and six gestational age-matched control (non-PE) placentas, using non-integrative Sendai virus-based method. I will assay their pluripotency, and characterize their epigenetic memory before and after reprogramming, evaluating both DNA methylation and histone H3 modifications. 2) I will characterize the trophoblast derived from both PE and control iPSCs, evaluating their ability to differentiate into both invasive EVT and multinucleated syncytiotrophoblast (STB). I will also evaluate both the gene expression and epigenetic profile of trophoblast derived from PE and non-PE iPSC. 3) Finally, I will test the effect of environmental factors, known to be involved in the pathogenesis of PE, on trophoblast derived from PE and non-PE iPSCs, including the effects of hypoxia, pro-inflammatory cytokines, and specific subpopulations of decidual leukocytes. The successful completion of this project will provide us with new models for studying this devastating disease, and thus the ability to generate novel diagnostic tools and therapeutic modalities in order to improve care for these women and babies.

项目摘要/摘要 胎盘是一个短暂的器官，在怀孕期间连接母亲和婴儿，并提供营养和 用于胎儿宫内生长的氧气。已知胎盘异常可导致先兆子痫 (PE)是影响2-8%妊娠的主要并发症，表现为新发高血压和 妊娠后期的蛋白尿。PE与生长受限和早产有关 在围产期和以后的生命中，这两者都与不良结局有关。许多 研究指出，胎盘上皮细胞(称为滋养层细胞)的异常分化是PE的病因。 特别是，侵袭性绒毛外滋养细胞(EVT)的发育和/或其功能在PE中受到影响， 导致浅着床和胎盘功能不全。然而，虽然这些都是被接受的 关于该病本质的假说，其潜在的病理生理机制主要仍未完全了解。 由于缺乏一个真正具有代表性的模式体系。在过去的十年里，方法的发展 产生诱导多能干细胞(IPSC)导致了对许多人类疾病进行建模的能力， 包括起源于心血管、神经系统和胰腺的那些。此外，在 同一时期，包括我们在内的多个群体已经表明，IPSC可以区分为 滋养层细胞，使用生长因子骨形态发生蛋白4(BMP4)。事实上，我自己的工作提供了 概念验证滋养层细胞分化缺陷，特别是与21三体相关的缺陷可以 使用IPSC进行建模。因此，我计划将这项技术应用于PE中的胎盘功能障碍模型， 以下三个目标：1)我将通过对脐带间充质干细胞进行重新编程来建立IPSCs (UC-MSCs)取自6个PE和6个胎龄匹配的对照(非PE)胎盘，使用非整合 仙台病毒为主的方法。我将分析它们的多能性，并在此之前描述它们的表观遗传记忆 在重新编程后，评估DNA甲基化和组蛋白H3修饰。2)我将描述 来自PE和对照IPSCs的滋养层细胞，评估它们分化为两者的能力 侵袭性EVT与多核合体滋养细胞(STB)我还将评估基因表达和 PE和非PE IPSC来源滋养层细胞的表观遗传学特征。3)最后，我要测试一下 已知参与PE发病机制的环境因素，对PE来源的滋养细胞和 非PE IPSCs，包括低氧、促炎细胞因子和特定亚群的影响 蜕膜白细胞。该项目的成功完成将为我们研究这一问题提供新的模型 毁灭性的疾病，因此有能力产生新的诊断工具和治疗方式 以改善对这些妇女和婴儿的护理。