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

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（一种妊娠高血压疾病）的潜在病因不是 众所周知，这种疾病始于浅胎盘和胎盘缺血， 过量的抗血管生成蛋白，如母亲的可溶性fms样酪氨酸激酶1（sFLT 1）， 血液中的血管内皮细胞是导致母体全身性内皮功能障碍的原因。自我更新 三维上皮类器官的结构和生理学非常相似， 原始器官已经成功地发展成各种组织类型， 多能干细胞（hiPSC）。然而，使用人胎盘滋养层细胞的类器官 hiPSC尚未产生。我们的目标是产生滋养层类器官 从疾病特异性hiPSC研究先兆子痫的发病机制，并筛选药物， 潜在的治疗目标我们将使用hiPSC产生滋养层类器官的新模型， 从正常妊娠和先兆子痫妊娠中复制妊娠早期， 到目前为止，研究人员大多无法获得的发展。在目标1中， 在我们的实验室中使用hiPSC衍生的滋养层细胞优化滋养层类器官方案 从供体成纤维细胞的分化方法，并将证实这些类器官的表型和 其功能类似于妊娠早期绒毛组织。然后，我们将测试假设，功能 来源于从早发性先兆子痫获得的hiPSC的滋养层类器官的能力将是 当与来自非高血压对照的滋养层类器官相比时受损。在目标2中， 我们将用人胎盘制备的因子在裸鼠中建立先兆子痫的母体综合征模型。 为了对人类先兆子痫进行建模，我们将使用来源于人的hiPSC产生滋养层类器官， 来自携带13三体胎儿的妇女的胎盘成纤维细胞， 由于13号染色体上sFLT 1基因的额外拷贝而导致先兆子痫的过度风险。然后我们将测试 靶向人sFLT 1-i14的独特C-末端的单克隆抗体的体内效力（ 灵长类特异性同种型）用于增强sFLT 1从体循环中的清除。由于 类器官的容易获得和复制早期发展阶段的能力， 特征细胞，滋养层类器官模型有望显着改善我们的研究。 了解先兆子痫，并提供测试潜在药物的快速筛选方法 并在产科中进一步推广精准医疗方法。我们的研究将对 不仅对先兆子痫的发病机制，而且对短期和长期的心血管疾病， 这些女性的并发症。