Placental Organoids to Model Preeclampsia

胎盘类器官模拟先兆子痫

• 批准号: 10594844
• 负责人: S. Ananth Karumanchi
• 金额: $ 41.75万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-15 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10594844
• 关键词: 3-Dimensional; Abnormal placentation; Activities of Daily Living; Address; Angiogenesis Inhibitors; Angiogenic Proteins; Binding; Biological Markers; Blood Circulation; Blood Vessels; Cardiovascular system; Cell Culture System; Cell Line; Cells; Cellular biology; Cerebral Palsy; Cessation of life; Chromosome 13; Circulation; Clinical; Complex; Complication; Data; Defect; Development; Developmental Process; Discipline of obstetrics; Disease; Drug Screening; Eclampsia; Embryonic Development; Endothelial Cells; Endothelial Growth Factors Receptor; Epithelium; Etiology; Event; Exhibits; FLT1 gene; Fetus; Fibroblasts; First Pregnancy Trimester; Genes; Genetic; Goals; Greater sac of peritoneum; HELLP Syndrome; Human; Hypertension; Hypoxia; Immunocompromised Host; Impairment; Implant; Invaded; Ischemia; Kidney Failure; LTK gene; Laboratories; Lead; Length; Maternal complication; Mediating; Membrane; Methods; Modeling; Monoclonal Antibodies; Mothers; Mus; Neonatal Mortality; Nervous System Trauma; Nude Mice; Organ; Organoids; Patau' s syndrome; Pathogenesis; Pathologic; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Physiology; Placenta; Placentation; Pre-Eclampsia; Pregnancy; Premature Birth; Primates; Protein Isoforms; Proteinuria; Protocols documentation; Rapid screening; Regenerative Medicine; Reproducibility; Research Personnel; Risk; Rodent; Seizures; Signal Transduction; Signs and Symptoms; Small for Gestational Age Infant; Specimen; Stroke; Structure; Syncytiotrophoblast; Syndrome; Testing; Time; Tissues; Uterus; Vascular Endothelial Growth Factor Receptor-1; Vascular Endothelial Growth Factors; Villous; Woman; cell type; comparison control; cytotrophoblast; early onset; endothelial dysfunction; epidemiology study; experimental study; fetal; functional disability; human model; improved; in vivo evaluation; induced pluripotent stem cell; infancy; neonatal morbidity; new therapeutic target; nonhuman primate; novel; novel therapeutics; precision medicine; pregnancy disorder; receptor; self-renewal; stem cell population; stem cells; therapeutic candidate; therapeutic evaluation; three dimensional cell culture; trophoblast; 3-Dimensional; Abnormal placentation; Activities of Daily Living; Address; Angiogenesis Inhibitors; Angiogenic Proteins; Binding; Biological Markers; Blood Circulation; Blood Vessels; Cardiovascular system; Cell Culture System; Cell Line; Cells; Cellular biology; Cerebral Palsy; Cessation of life; Chromosome 13; Circulation; Clinical; Complex; Complication; Data; Defect; Development; Developmental Process; Discipline of obstetrics; Disease; Drug Screening; Eclampsia; Embryonic Development; Endothelial Cells; Endothelial Growth Factors Receptor; Epithelium; Etiology; Event; Exhibits; FLT1 gene; Fetus; Fibroblasts; First Pregnancy Trimester; Genes; Genetic; Goals; Greater sac of peritoneum; HELLP Syndrome; Human; Hypertension; Hypoxia; Immunocompromised Host; Impairment; Implant; Invaded; Ischemia; Kidney Failure; LTK gene; Laboratories; Lead; Length; Maternal complication; Mediating; Membrane; Methods; Modeling; Monoclonal Antibodies; Mothers; Mus; Neonatal Mortality; Nervous System Trauma; Nude Mice; Organ; Organoids; Patau' s syndrome; Pathogenesis; Pathologic; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Physiology; Placenta; Placentation; Pre-Eclampsia; Pregnancy; Premature Birth; Primates; Protein Isoforms; Proteinuria; Protocols documentation; Rapid screening; Regenerative Medicine; Reproducibility; Research Personnel; Risk; Rodent; Seizures; Signal Transduction; Signs and Symptoms; Small for Gestational Age Infant; Specimen; Stroke; Structure; Syncytiotrophoblast; Syndrome; Testing; Time; Tissues; Uterus; Vascular Endothelial Growth Factor Receptor-1; Vascular Endothelial Growth Factors; Villous; Woman; cell type; comparison control; cytotrophoblast; early onset; endothelial dysfunction; epidemiology study; experimental study; fetal; functional disability; human model; improved; in vivo evaluation; induced pluripotent stem cell; infancy; neonatal morbidity; new therapeutic target; nonhuman primate; novel; novel therapeutics; precision medicine; pregnancy disorder; receptor; self-renewal; stem cell population; stem cells; therapeutic candidate; therapeutic evaluation; three dimensional cell culture; trophoblast

While the underlying etiology of preeclampsia (a hypertensive disorder of pregnancy) is not known, the disease starts with shallow placentation and placental ischemia which in turn releases excess of anti-angiogenic proteins such as soluble fms-like tyrosine kinase 1 (sFLT1) in the mother's bloodstream that is responsible for the systemic maternal endothelial dysfunction. Self-renewing three-dimensional epithelial organoids that closely resemble the structure and physiology of the original organ have been successfully developed into various tissue types using human induced pluripotent stem cells (hiPSCs). However, organoids of the human placental trophoblasts using hiPSCs are yet to be generated. Our goal of this proposal is to generate trophoblast organoids from disease-specific hiPSCs to study preeclampsia pathogenesis and to screen for drugs as potential treatment targets. We will generate a new model of trophoblast organoid using hiPSCs, replicating the early stage of gestation from normal and preeclamptic pregnancies, a time in development that has – until now – has been mostly inaccessible to researchers. In aim 1, we will optimize trophoblast organoid protocols in our laboratory using hiPSCs derived trophoblast differentiation method from donor fibroblasts and will confirm that these organoids phenotypically and functionally behave like first trimester villous tissue. We will then test the hypothesis that the functional capacity of trophoblast organoids derived from hiPSCs obtained from early-onset preeclampsia will be impaired when compared to trophoblast organoids derived from non-hypertensive controls. In aim 2, we will model maternal syndrome of preeclampsia in nude mice with factors made by human placenta. To model human preeclampsia, we will generate trophoblast organoids using hiPSCs derived from placental fibroblasts from women carrying a fetus with trisomy 13, a disorder characterized by 10-fold excess risk of preeclampsia due to extra copy of sFLT1 gene on chromosome 13. We will then test in vivo efficacy of monoclonal antibodies that target the unique C-terminus of human sFLT1-i14 (the isoform that is primate-specific) for enhanced clearance of sFLT1 from systemic circulation. Due to the organoid's ready access and ability to replicate the early stages of development from well- characterized cells, the trophoblast organoid model promises to significantly improve our understanding of preeclampsia and provides rapid screening methods for testing potential drugs and furthering precision medicine methods in obstetrics. Our studies will have major implications not only for the pathogenesis of preeclampsia, but also for short and long-term cardiovascular complication in these women.

虽然先兆子痫的潜在病因（一种高血压疾病）不是 已知，该疾病始于浅置位和占地缺血，从而又释放 过量的抗血管生成蛋白，例如固体FMS样酪氨酸激酶1（SFLT1） 负责全身材料内皮功能障碍的血液。自我更新 三维上皮器官非常类似于 使用人类诱导的原始器官已成功地发展为各种组织类型 多能干细胞（HIPSC）。但是，使用人类斑点滋养细胞的器官使用 HIPSC尚未生成。该提议的目标是产生滋养细胞类器官 从疾病特异性的HIPSC来研究前肿瘤的发病机理，并筛查药物 潜在的治疗目标。我们将使用hipscs生成一种新的滋养细胞器官的模型， 复制正常和先兆怀孕的妊娠早期阶段， 到目前为止，研究人员的发展大多是无法访问的。在AIM 1中，我们将 使用HIPSC衍生的滋养细胞优化实验室中的滋养细胞器官方案 与供体成纤维细胞的分化方法，将在表型和 在功能上的行为就像头三个月的绒毛组织。然后，我们将测试功能性的假设 从早期发作前启示性获得的HIPSC得出的滋养细胞类器官的能力将是 与从非高血压对照中得出的滋养细胞类器官相比，受损。在AIM 2中， 我们将在裸小鼠中的前美氏症的母体综合征与人类心papeta所产生的因素进行建模。 为了建模人类前启示识别，我们将使用来自 来自胎儿的胎盘成纤维细胞，这些胎儿带有三体性胎儿，这种疾病为特征10倍 由于染色体13上的SFLT1基因副本，因此先兆子痫的风险过多。然后，我们将进行测试 靶向人类sflt1-i14的独特C末端的单克隆抗体的体内效率（ 具有特定于私有的同工型），以增强系统循环的SFLT1清除率。由于 Organoid的即时访问和能力复制开发的早期阶段 特征细胞的滋养细胞有机模型有望显着改善我们的 了解先兆子痫并提供快速筛选方法来测试潜在药物 以及进一步的精确医学方法。我们的研究将具有重大影响 不仅对于先兆子痫的发病机理，而且对于短期和长期心血管 这些女性的并发症。