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（一种妊娠期高血压疾病）的潜在病因并不明确。 众所周知，这种疾病始于浅胎盘和胎盘缺血，进而释放 母亲体内过量的抗血管生成蛋白，例如可溶性 fms 样酪氨酸激酶 1 (sFLT1) 负责全身母体内皮功能障碍的血液。 三维上皮类器官，其结构和生理学非常类似于 利用人体诱导，原始器官已成功发育成各种组织类型 然而，人类胎盘滋养层的类器官。 hiPSC 尚未生成。我们该提案的目标是生成滋养层类器官。 从疾病特异性 hiPSC 中研究先兆子痫发病机制并筛选药物 我们将使用 hiPSC 生成一种新的滋养层类器官模型， 复制正常妊娠和先兆子痫妊娠的妊娠早期阶段， 到目前为止，研究人员基本上无法实现这一目标 1。 在我们的实验室中使用 hiPSC 衍生的滋养层优化滋养层类器官方案 与供体成纤维细胞的分化方法，并将确认这些类器官的表型和 功能上类似于妊娠早期绒毛组织，然后我们将检验功能性假设。 从早发性先兆子痫中获得的 hiPSC 衍生的滋养层类器官的能力将 与来自非高血压对照的滋养层类器官相比，其受损。 在目标 2 中， 我们将用人胎盘制造的因子在裸鼠中模拟先兆子痫的母体综合症。 为了模拟人类先兆子痫，我们将使用来自以下来源的 hiPSC 生成滋养层类器官： 来自怀有 13 三体症胎儿的女性的胎盘成纤维细胞，这是一种以 10 倍染色体异常为特征的疾病 由于 13 号染色体上 sFLT1 基因的额外拷贝，导致先兆子痫的风险过高。然后我们将在 靶向人 sFLT1-i14 独特 C 末端的单克隆抗体的体内功效（ 灵长类动物特异性的亚型）可增强 sFLT1 从体循环中的清除率。 类器官的现成访问能力和复制早期发育阶段的能力 滋养层类器官模型有望显着改善我们的特征细胞 了解先兆子痫并提供测试潜在药物的快速筛选方法 我们的研究将产生重大影响。 不仅针对子痫前期的发病机制，而且针对短期和长期心血管疾病 这些女性的并发症。