Placental Organoids for Modeling and Treating Preeclampsia

用于建模和治疗先兆子痫的胎盘类器官

• 批准号: 10464766
• 负责人: S. Ananth Karumanchi
• 金额: $ 4.9万
• 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: 2022-09-22 至 2022-12-17
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10464766
• 关键词: 3-Dimensional; Abnormal placentation; Activities of Daily Living; Address; Angiogenic Proteins; Binding; Blood Circulation; Blood Vessels; Cardiovascular system; Cell Culture System; Cell Line; Cells; Cellular biology; Cerebral Palsy; Cessation of life; Chromosome 13; Clinical; Data; Defect; Development; Developmental Process; Discipline of obstetrics; Disease; Drug Screening; Eclampsia; Embryonic Development; Endothelial Growth Factors Receptor; Epithelial; 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; 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; Pharmaceutical Preparations; Phenotype; Physiology; Placenta; Placentation; Plant Roots; Population; 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; biomarker development; cell type; 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 3D 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 from 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.

先兆子痫（一种妊娠期高血压疾病）的潜在病因 目前尚不清楚，该疾病始于浅胎盘和胎盘缺血， 转释放过量的抗血管生成蛋白，例如可溶性 fms 样酪氨酸激酶 1 (sFLT1) 在母亲的血液中，负责全身母体内皮细胞 自我更新的 3D 上皮类器官，其结构和功能非常相似。 原始器官的生理学已成功发育成各种组织 然而，使用人类诱导多能干细胞（hiPSC）的类器官。 使用 hiPSC 的人胎盘滋养层尚未产生，我们的目标尚未实现。 提议是从疾病特异性 hiPSC 中生成滋养层类器官来进行研究 先兆子痫的发病机制并筛选药物作为潜在的治疗靶点。 将使用 hiPSC 生成一个新的滋养层类器官模型，复制早期的 正常妊娠和先兆子痫妊娠的妊娠阶段，是一个发育时期 到目前为止，研究人员基本上无法访问。在目标 1 中，我们将进行优化。 我们实验室使用 hiPSC 衍生的滋养层的滋养层类器官方案 与供体成纤维细胞的分化方法将确认这些类器官 表型和功能与妊娠早期绒毛组织相似，然后我们将对其进行测试。 假设来自 hiPSC 的滋养层类器官具有功能能力 与滋养层类器官相比，早发性先兆子痫的类器官会受到损害 来自非高血压对照 在目标 2 中，我们将模拟孕产妇综合症。 用人胎盘制造的因子在裸鼠中建立先兆子痫模型。 先兆子痫，我们将使用源自胎盘的 hiPSC 生成滋养层类器官 来自携带 13 三体胎儿的女性的成纤维细胞，这是一种以 10 倍体型为特征的疾病 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。我们将 然后测试针对人类独特 C 末端的单克隆抗体的体内功效 sFLT1-i14（灵长类特异性亚型），用于增强 sFLT1 的清除 由于类器官易于进入并具有复制早期的系统能力。 充分表征的细胞的发育阶段，滋养层类器官模型 有望显着提高我们对先兆子痫的了解，并提供快速 测试潜在药物的筛选方法和推进精准医学方法 在产科。