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：
7055298
负责人：
DIANA W. BIANCHI
金额：
$ 31.93万
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

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）：我们要求支持以下假设：胎儿细胞是通过怀孕生理学的，并在流产，流产或分娩后保留在成年人类女性中，涵盖了我们称为“怀孕相关的祖细胞（Papc）的新型细胞”。迄今为止，围绕成年干细胞可塑性的争议几乎忽略了怀孕在女性中的作用。 PAPC，如果证明是真正的干细胞，将具有起源的发育优势，但可以在没有以前怀孕的成年女性的道德上争议的情况下检索。我们在人类成年人，非转移的雌性中拥有广泛的初步数据，该胎儿细胞（根据Y染色体和胎儿特异性的DNA多态性鉴定）通过妊娠获得，可在外周血和临床疾病的器官中检测到，并具有多局部性。由于有必要从活检和/或尸检标本中获得临床材料，因此这些研究具有描述性，而不是机械性的。我们的假设将在动物模型，一种表达绿色荧光蛋白（GFP）的转基因雄性小鼠或繁殖到野生型雌性小鼠的荧光素酶。这将使我们能够控制生殖历史，并检验有关母体胎儿细胞的可塑性和活性的多种假设。 GFP和荧光素酶序列是主要的转基因。一半的胎儿幼崽将携带转基因，并根据构建体在其某些或所有细胞中表达绿色荧光标记。胎儿细胞荧光绿色，可以使用各种技术在母体组织中识别和跟踪，包括体内全动物成像，荧光显微镜和实时PCR扩增。在特定目的1中，我们将测试以下假设：特定因素会影响母亲中胎儿细胞微chi（FCMC）的发展。在特定的目标2中，我们将使用化学，外科手术，遗传和缺血模型来确定在特定的组织损伤场景中是否募集胎儿细胞，并通过分析整体健康状况和寿命，靶心器官功能，伤口愈合的差异和差异基因表达来促进孕产妇损伤的修复。在特定目标3中，我们将检查鼠微chimeric胎儿细胞的细胞表面特征，并进行微阵列分析，以确定FCMC是否是由于1或多种细胞类型引起的。在特定目标4中，我们将检验以下假设：胎儿干细胞比成年干细胞具有优势，并有助于长期生存或改善器官功能。长期目标是确定怀孕是否通过获得具有治疗潜力的独特细胞来赋予女性长期优势。