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.

虽然先兆子痫（一种妊娠期高血压疾病）的潜在病因 尚不清楚，这种疾病始于浅胎盘和胎盘缺血， turn会释放过量的抗血管生成蛋白，如可溶性fms样酪氨酸激酶1 （sFLT 1）在母亲的血液，负责全身母体内皮细胞 功能障碍自我更新的3D上皮类器官与结构非常相似， 原始器官的生理机能已经成功地发育成各种组织 使用人类诱导多能干细胞（hiPSC）的类型。然而， 使用hiPSC的人胎盘滋养层细胞还有待产生。我们的目标是 一项提案是从疾病特异性hiPSC中产生滋养层类器官，以研究 子痫前期发病机制和筛选药物作为潜在的治疗目标。我们 将使用hiPSC产生一种新的滋养层类器官模型， 正常妊娠和先兆子痫妊娠的妊娠阶段， 到目前为止，研究人员还无法接触到。在目标1中，我们将优化 在我们的实验室中使用hiPSC衍生的滋养层细胞的滋养层类器官方案 从供体成纤维细胞分化的方法，并将确认这些类器官， 在表型和功能上表现得像妊娠早期绒毛组织。然后我们将测试 假设从hiPSC获得的滋养层类器官的功能能力 与滋养层类器官相比， 来自非高血压对照。在目标2中，我们将模拟 用人胎盘制备的因子对裸鼠进行子痫前期实验。来模拟人类 在先兆子痫中，我们将使用来自胎盘的hiPSC产生滋养层类器官。 来自携带13三体胎儿的妇女的成纤维细胞，13三体是一种特征为10倍染色体缺失的疾病， 由于13号染色体sFLT 1基因的额外拷贝而导致先兆子痫的过度风险。我们将 然后测试靶向人免疫球蛋白的独特C-末端的单克隆抗体的体内功效， sFLT 1-i14（灵长类特异性同种型），用于增强sFLT 1从 体循环由于类器官容易进入并能够复制早期的 发育阶段从良好表征的细胞，滋养层类器官模型 有望显著提高我们对先兆子痫的理解，并提供快速的 用于测试潜在药物的筛选方法和进一步的精确医学方法 在产科。