Aneuploid Cells in the Human Placenta

人胎盘中的非整倍体细胞

• 批准号: 6824639
• 负责人: Heinz-Ulrich Guenter Weier
• 金额: $ 32.54万
• 依托单位: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-15 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6824639
• 关键词: aneuploidy; bromodeoxyuridine; cell differentiation; cell population study; cell proliferation; clinical research; confocal scanning microscopy; cytogenetics; embryo implantation; fluorescent in situ hybridization; gene expression; histocompatibility gene; histology; human subject; immunocytochemistry; karyotype; mammalian embryology; patient oriented research; phenotype; placenta; pregnancy; three dimensional imaging /topography; trisomy; trophoblast

DESCRIPTION (provided by applicant): In eutherian mammals, proper fetal development depends on a functional placenta. Specialized placental compartments anchor the embryo to the uterus and maintain a steady supply of oxygen and nutrients to the embryo, while other parts manage the disposal of metabolites. Two placental structures facilitate these functions: floating and anchoring villi. The genetic makeup of floating villi has been studied in detail. Little is known about the genetic make-up of anchoring villi, but our cytogenetic studies of fetal cells isolated from anchoring villi or the uterine wall revealed an unexpected large fraction of invading cytotrophoblasts (CTBs) that carry numerical chromosomal aberrations. We postulate that aneuploidy at the fetal-maternal interface is an integral part of the normal placentation program. We hypothesize that in the absence of mutations, genomic changes in CTBs such as gains or losses of chromosomes are normal cellular mechanisms that limit the proliferative as well as the invasive capabilities of CTBs. To test our hypothesis, we propose to investigate the karyotype of CTBs as well as phenotypic markers of cell proliferation or differentiation along an invasive CTB phenotype. In the study proposed here, investigators with complementary expertise in early embryonic development, placental function, molecular cytogenetics and digital image processing will collaborate to localize aneuploid cells in different placental compartments. First, we will determine the types of numerical chromosome abnormalities in the uterine wall and in anchoring as well as floating villi in normal placental specimens and placental tissues carrying a trisomy 21. Next, we will determine the spatial distribution of genomic instability in normal and trisomic second trimester placentas. Techniques used will be fluorescence in situ hybridization (FISH), confocal microscopy and 3D image reconstruction of thick tissue sections to localize aneuploid cells in their histological context. Finally, we will determine the effects of aneuploidy on the proliferative capacity of CTBs as measured by BrdU incorporation and expression of the HLA-G gene, an important marker for CTB differentiation along the invasive pathway. At the end of this project, we will have developed the technology to accurately score all 24 human chromosomes in interphase cells and demonstrated the feasibility to localize aneuploid cells in 3-dimension in thick tissue sections. We will also have detailed information about the frequency, localization and types of aneuploid cells at the fetal-maternal interface and the effects of aneuploidy on CTB proliferation and HLA-G gene expression.

描述（由申请人提供）：在Eutherian哺乳动物中，适当的胎儿发育取决于功能性胎盘。专门的胎盘室将胚胎固定在子宫上，并将氧气和养分稳定供应到胚胎，而其他部分则管理代谢物的处置。两个胎盘结构有助于这些功能：浮动和锚定绒毛。详细研究了漂浮绒毛的基因组成。关于锚定绒毛的遗传组成知之甚少，但是我们从锚固绒毛或子宫壁分离的胎儿细胞的细胞遗传学研究表明，出乎意料的大部分入侵的细胞增生细胞细胞（CTB），这些细胞细胞（CTB）带有数值染色体差异。我们假设胎儿​​界面处的非整倍性是正常胎盘程序的组成部分。我们假设在没有突变的情况下，CTB的基因组变化（例如增益或染色体损失）是正常的细胞机制，限制了CTB的增殖和侵入性能力。为了检验我们的假设，我们建议研究CTB的核型，以及沿着侵入性CTB表型的细胞增殖或分化的表型标记。在此处提出的研究中，具有早期胚胎发育，胎盘功能，分子细胞遗传学和数字图像处理方面具有互补专业知识的研究人员将协作以将非整倍体细胞定位在不同的胎盘室中。首先，我们将确定子宫壁中的数字染色体异常类型，锚定以及浮动绒毛在正常胎盘标本和带有三体术的胎盘组织中。接下来，我们将确定正常和三体性基因组不稳定性的空间分布妊娠胎盘。所使用的技术将是荧光原位杂交（FISH），共聚焦显微镜和厚组织切片的3D图像重建，以在其组织学环境中定位非整倍性细胞。最后，我们将确定非整倍性对CTB的增殖能力的影响，如HLA-G基因的BRDU掺入和表达，这是CTB沿侵入性途径的重要标记。在该项目结束时，我们将开发该技术，以准确地在相间细胞中准确评分所有24个人类染色体，并证明在厚组织切片中将非整倍体细胞定位在三维细胞中的可行性。我们还将获得有关胎儿界面处的频率，定位和类型的频率，定位和类型，以及非整倍性对CTB增殖和HLA-G基因表达的影响。