The Role of Fetal Cell Microchimerism in Maternal Repair

胎儿细胞微嵌合在母体修复中的作用

基本信息

批准号：
7341612
负责人：
DIANA W. BIANCHI
金额：
$ 30.39万
依托单位：
TUFTS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-04-15 至 2010-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7341612
关键词：
Adult Affect Age Animal Model Animals Autopsy Biological Assay Biopsy Blood Circulation Breeding Carbon Tetrachloride Cell Separation Cell surface Cells Characteristics Chemical Models Chemicals Chronic Clinical Collagen Color Coronary artery Culture Techniques DNA Data Development Endothelial Cells Female Flow Cytometry Fluorescence Microscopy Fumarylacetoacetase Deficiency Disease Genes Genetic Genetic Polymorphism Green Fluorescent Proteins Healed Health Heart Hematopoietic stem cells Hepatic Histology Human Image Injection of therapeutic agent Injury Kidney Knock-out Ligation Liver Liver diseases Longevity Luciferases Mesenchymal Stem Cells Methods Microarray Analysis Microchimerism Modeling Mothers Mus Operative Surgical Procedures Organ Organ Survival Partial Hepatectomy Patient currently pregnant Personal Satisfaction Phenotype Placentation Polymerase Chain Reaction Population Pregnancy Process Property Rate Recording of previous events Recruitment Activity Reproductive History Research Role Side Skin Sorting - Cell Movement Specimen Spontaneous abortion Staining method Stains Stem cells Surface Antigens Surgical Injuries Techniques Testing Therapeutic Time Tissue-Specific Gene Expression Tissues Transgenes Transgenic Organisms Wild Type Mouse Woman Wound Healing Y Chromosome abortion adult stem cell base cell motility cell type enzyme deficiency fetal fetal stem cell fetus cell fluorescence imaging healing human female improved in vivo injury and repair liver cell proliferation male novel peripheral blood pup repaired response response to injury trafficking wound

项目摘要

DESCRIPTION (provided by applicant): We are requesting support to test the hypothesis that fetal cells, acquired physiologically through pregnancy, and retained in the adult human female following abortion, miscarriage, or delivery, encompass a novel population of cells that we have termed the "Pregnancy-Associated Progenitor Cell (PAPC)." To date, the controversy surrounding the plasticity of adult stem cells has virtually ignored the role of pregnancy in females. PAPCs, if shown to be true stem cells, would have the developmental advantages of being fetal in origin yet could be retrieved without ethical controversy from an adult female who has previously been pregnant. We have extensive preliminary data in the human adult, non-transfused female that fetal cells (identified on the basis of the Y chromosome as well as fetal-specific DNA polymorphisms), acquired through pregnancy, are detectable in peripheral blood and clinically diseased organs, and have multi-lineage capacity. Due to the necessity of obtaining clinical material from biopsy and/or autopsy specimens, these studies have been descriptive and not mechanistic. Our hypothesis will be tested in an animal model, a transgenic male mouse expressing either green fluorescent protein (GFP) or luciferase bred to wild-type female mice. This will allow us to control reproductive histories, and test multiple hypotheses regarding the plasticity and activity of fetal cells in the maternal body. The GFP and luciferase sequences are dominant transgenes. Half of the fetal pups will carry the transgene, and express the green fluorescent marker in some or all of their cells, depending on the construct. Fetal cells fluoresce green and can be identified and tracked in maternal tissues using a variety of techniques, including in vivo whole animal imaging, fluorescence microscopy, and real-time PCR amplification. In specific aim 1 we will test the hypothesis that specific factors affect the development of fetal cell microchimerism (FCMC) in the mother. In specific aim 2 we will use chemical, surgical, genetic, and ischemic models to determine if fetal cells are recruited in specific tissue injury scenarios and contribute to the repair of maternal injury by analyzing overall well being and longevity, target organ function, differences in wound healing, and differential gene expression. In specific aim 3 we will examine the cell surface characteristics of the murine microchimeric fetal cells and perform microarray analysis to determine whether FCMC is due to 1 or multiple cell types. In specific aim 4 we will test the hypothesis that fetal stem cells have an advantage over adult stem cells and contribute to prolonged survival or improved organ function. The long-term objective is to determine if pregnancy confers a long-term advantage to a female by resulting in the acquisition of unique cells that have therapeutic potential.

描述（由申请人提供）：我们请求支持来测试以下假设：胎儿细胞通过怀孕获得生理学并在流产、流产或分娩后保留在成年女性体内，包含一个新的细胞群，我们将其称为“妊娠相关祖细胞（PAPC）。”迄今为止，围绕成体干细胞可塑性的争议实际上忽略了女性怀孕的作用。如果证明 PAPC 是真正的干细胞，它将具有胎儿起源的发育优势，而且可以从以前怀孕的成年女性身上回收，而不会引起伦理争议。我们在未输血的成年女性中拥有大量初步数据，表明通过怀孕获得的胎儿细胞（根据 Y 染色体以及胎儿特异性 DNA 多态性进行鉴定）可在外周血和临床患病器官中检测到，并具有多谱系能力。由于需要从活检和/或尸检标本中获取临床材料，这些研究是描述性的而不是机械性的。我们的假设将在动物模型中进行测试，该模型是与野生型雌性小鼠交配的表达绿色荧光蛋白（GFP）或荧光素酶的转基因雄性小鼠。这将使我们能够控制生殖历史，并测试有关母体内胎儿细胞的可塑性和活性的多种假设。 GFP 和荧光素酶序列是显性转基因。一半的胎儿幼崽将携带转基因，并在其部分或全部细胞中表达绿色荧光标记，具体取决于构建体。胎儿细胞发出绿色荧光，可以使用多种技术在母体组织中进行识别和追踪，包括体内整体动物成像、荧光显微镜和实时 PCR 扩增。在具体目标 1 中，我们将检验特定因素影响母亲胎儿细胞微嵌合体 (FCMC) 发育的假设。在具体目标 2 中，我们将使用化学、手术、遗传和缺血模型来确定胎儿细胞是否在特定组织损伤情况下被招募，并通过分析整体健康和寿命、靶器官功能、母体损伤的差异来促进母体损伤的修复。伤口愈合和差异基因表达。在具体目标 3 中，我们将检查鼠微嵌合胎儿细胞的细胞表面特征，并进行微阵列分析以确定 FCMC 是否由 1 种或多种细胞类型引起。在具体目标 4 中，我们将检验以下假设：胎儿干细胞比成体干细胞具有优势，有助于延长生存时间或改善器官功能。长期目标是确定怀孕是否会通过获得具有治疗潜力的独特细胞而为女性带来长期优势。