Modeling Preeclampsia Using Human Pluripotent Stem Cells

使用人类多能干细胞模拟先兆子痫

基本信息

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
项目类别：
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

项目摘要

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/摘要胎盘是一种瞬态器官，在怀孕期间连接妈妈和婴儿，并提供营养和宫内胎儿生长的氧气。众所周知，异常胎盘会导致先兆子痫（PE），一个主要的并发症影响了所有怀孕的2-8％，以新的发作高血压和怀孕后半段的蛋白尿。 PE与生长限制和早产相关宝贝，两者都与不良后果有关，无论是在围产期时期和生命之后。许多研究指出，胎盘上皮细胞的异常分化，称为滋养细胞，为PE的病因。具体而言，在PE中，侵入性跨性滋养细胞（EVT）和/或其功能的发展受到影响，导致浅植入和胎盘不足。但是，虽然这些被接受关于疾病性质的假设，基本的病理生理学仍然没有完全理解由于缺乏真正的代表性模型系统。在过去的十年中，发展的方法产生诱导的多能干细胞（IPSC）已导致对许多人类疾病进行建模的能力，包括起源于心血管和神经系统以及胰腺的人。另外，在同一时间段，包括我们的多个组，表明IPSC可以区分为滋养细胞，使用生长因子，骨形态发生蛋白4（BMP4）。我自己的工作实际上提供了概念证明滋养细胞分化的缺陷，特别是与三体性相关的缺陷，可以使用IPSC进行建模。因此，我计划将此技术应用于PE中的胎盘功能障碍，并使用以下三个目标：1）我将通过重新编程脐带间充质干细胞来建立IPSC （UC-MSC）使用非整合性基于仙台病毒的方法。我将分析他们的多能性，并在表观遗传记忆之前表征重新编程后，评估DNA甲基化和组蛋白H3修饰。 2）我会描述源自PE和控制IPSC的滋养细胞，评估其分化为两者的能力侵入性EVT和多核合成毒剂细胞（STB）。我还将评估基因表达和源自PE和非PE IPSC的滋养细胞的表观遗传谱。 3）最后，我将测试已知与PE的发病机理有关的环境因素在PE和PE的滋养细胞上非PE IPSC，包括缺氧的影响，促炎性细胞因子和特定亚群决结物细胞。该项目的成功完成将为我们提供研究的新模型毁灭性疾病，因此具有生成新颖的诊断工具和治疗方式的能力改善对这些妇女和婴儿的护理。