p63-Dependent Checkpoints in Oocytes

卵母细胞中 p63 依赖性检查点

基本信息

批准号：
7900949
负责人：
FRANK D. MCKEON
金额：
$ 48.9万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-01 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7900949
关键词：
Address Apoptotic BH3 Domain Cell Death Cells Cessation of life Chemotherapy-Oncologic Procedure Chromosomes Congenital Abnormality DNA Binding DNA Damage Data Development Dose Embryo Embryonic Development Ensure Equilibrium Exposure to Failure Family Female Female infertility Fertility Fertilization Gene Duplication Gene Expression Gene Targeting Genes Genetic Genetic Recombination Genomics Homologous Gene Human Infertility Ionizing radiation Knockout Mice Lead Link MLH1 gene Mammals Mediating Mediator of activation protein Meiosis Modeling Mus Mutant Strains Mice Mutation Oocytes Orthologous Gene Ovarian Ovary Ovulation Pattern Pharmaceutical Preparations Phenotype Phosphorylation Play Protein p53 Proteins Puma Radiation Role SPO11 gene Signal Pathway Site Somatic Cell Staging TP53 gene Testing Therapeutic Tumor Suppression Woman Work base cancer prevention chemotherapy egg fly implantation insight irradiation knockout gene mouse model mutant null mutation postnatal prevent programs response sickling

项目摘要

Meiosis in the female germline of mammals is marked by an exceedingly prolonged arrest in meiosis I between homologous chromosome recombination and ovulation, which in humans can last five decades. During this arrest period, oocytes are highly vulnerable to DNA damage from endogenous, environmental, or therapeutic origins. How DNA damage triggers the death of these arrested oocytes is poorly understood but we now know that p63, a homolog of the p53 tumor suppressor, is highly expressed in these oocytes and plays an essential role in their death following exposure to ionizing radiation. These findings, based on mouse gene knockout models, have raised fundamental questions regarding the strategies to balance female fertility with the need for preventing mutations in a limited pool of oocytes. These questions are addressed in two specific aims. Aim 1 is to determine if p63 also functions to eradicate oocytes damaged by the highly mutagenic steps of homologous chromosome recombination. We also want to determine if TAp63 also contributes to oocyte death in young women undergoing chemotherapy for cancer. Our preliminary data for Aim 1 support the concept that TAp63 functions both in a post-pachytene checkpoint and in oocyte death due to chemotherapeutic drugs linked to ovarian failure. In this aim we will assess the ability of TAp63 null mutations in mice to rescue oocyte death that accompanies meiosis in the absence of key genes such as ATM, DCM, MHL, and SPO11 using specific mouse mutants. Aim 2 is to identify the transcriptional targets of TAp63 protein in oocytes following lethal and sub-lethal doses of ionizing radiation, chemotherapy, and homologous chromosome recombination. Our preliminary data strongly implicates the proapoptotic Puma gene in oocyte death, and the role of this gene in oocyte death will be probed in mouse knockout models. We anticipate that these studies will reveal much about how the oocyte maintains a high degree of genomic fidelity, how chemotherapy leads to female infertility, and insights into how the p53 family acts to eliminate cells with DNA damage to promote tumor suppression.

哺乳动物女性种系中的减数分裂是在减数分裂中延长的延长的标志在同源染色体重组和排卵之间，在人类中可以持续五个几十年。在此逮捕期间，卵母细胞高度容易受到内源性的DNA损害，环境或治疗起源。 DNA损伤如何触发这些被捕的卵母细胞的死亡知之甚少，但我们现在知道p63是p53肿瘤抑制剂的同源物在这些卵母细胞中表达，并在暴露于电离后死亡中起着至关重要的作用辐射。这些基于鼠标基因敲除模型的发现提高了基本关于平衡女性生育能力与预防突变的策略的问题有限的卵母细胞。这些问题以两个具体目标解决。目标1是确定如果p63还起作用，可以消除受同源的高度诱变步骤损坏的卵母细胞染色体重组。我们还想确定TAP63是否也导致卵母细胞死亡年轻女性接受癌症化学疗法。我们AIM 1的初步数据支持 TAP63在Pachytene检查点和卵母细胞死亡中起作用的概念与卵巢衰竭有关的化学治疗药物。在此目标中，我们将评估TAP63 NULL的能力在没有关键基因的情况下，小鼠的突变挽救了伴随减数分裂的卵母细胞死亡例如使用特定的小鼠突变体等ATM，DCM，MHL和SPO11。目标2是确定致命和亚特剂量电离后，卵母细胞中TAP63蛋白的转录靶标放射，化学疗法和同源染色体重组。我们的初步数据强烈暗示了卵母细胞死亡中的凋亡PUMA基因，该基因在卵母细胞死亡中的作用将会在鼠标敲除模型中进行探测。我们预计这些研究将大大揭示卵母细胞保持高度的基因组保真度，化学疗法如何导致女性不育症，并深入了解p53家族如何消除具有DNA损伤的细胞以促进肿瘤抑制。