Molecular Basis of Aneuploidy

非整倍体的分子基础

基本信息

批准号：
7576183
负责人：
DEBANANDA PATI
金额：
$ 23.27万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-03-07 至 2011-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7576183
关键词：
Address Affect Agreement Anaphase Aneuploidy Binding Biological Models Breast Breast Cancer Model Cancer Biology Cell Culture Techniques Cell Line Cells Chromosomal Instability Chromosomal Stability Chromosome Segregation Chromosome abnormality Chromosomes Chronic Cleaved cell Complementary DNA Contralateral Data Development Diploidy Elements Endopeptidases Epithelial Cells Estrogens Fatty acid glycerol esters Gene Expression Genes Genetic Transcription Genotype Goals Gonadal Steroid Hormones Hormonal Hormone Responsive Hormones Human Human Characteristics Inbred BALB C Mice Incidence Indium Injection of therapeutic agent Knockout Mice Laboratories Lead Malignant Neoplasms Mammary Neoplasms Mammary Tumorigenesis Mammary gland Measures Messenger RNA Metaphase Mitosis Mitotic Modeling Molecular Mus Mutation Palpation Pathogenesis Peptide Hydrolases Physiological Pituitary Isograft Play Ploidies Preparation Progesterone Promoter Regions Protein p53 Proteins Publishing Regulation Research Personnel Risk Risk Factors Role Sequence Analysis Set protein Signal Transduction Sister Chromatid Steroids Structure System TP53 gene Techniques Testing Therapeutic Intervention Transcriptional Regulation Transgenic Mice Transplantation Tumor Suppressor Proteins Tumorigenicity Validation Wild Type Mouse base cancer cell carcinogenesis cohesin cohesion in vivo in vivo Model innovation malignant breast neoplasm mouse model mutant neoplastic cell overexpression programs promoter research study segregation separase steroid hormone tumor tumor progression vector control

项目摘要

This proposal focuses on an important but unexplored problem - howsteroid hormones, which are well known risk factors, interact with p53 mutations to produce aneuploidy and malignancy, and how the chromosomal segregation protein Separase is involved? Our sex-steroid dependent p53-mice preneoplastic breast cancer model allows a unique approach to this problem. In this model, steroidal induction in p53 mutant mammary glands results in chromosomal instability, aneuploidy and tumor formation analogous to that seen in majority of human breast cancers. We propose a paradigm that there is a set of proteins whose deregulation promotes aneuploidy (termed PRAN; Promoter of Aneuploidy) including chromosomal instability which results in loss or gain of whole or parts of chromosomes, and that PRAN proteins are interactively regulated by steroid hormones and the tumor suppressor p53. Our published and new preliminary data provide the first evidence that steroid hormones play a role in the regulation of mitotic proteins involved in sister chromatid cohesion and separation. We propose that the combined effect of mutation of the tumor suppressor p53 and signaling by steroid hormones produces aneuploidv in breast cancer by affecting expression of key proteins involved in chromosomal separation. We focuses on the elements that regulate chromosomal segregation, particularly sister chromatid cohesion/separation proteins, as candidate PRAN proteins, since chromosome missegregation during mitosis can lead to aneuploidy. A key gene in this analysis is ESPL1, which encodes an endopeptidase called Separase that separates joined sister chromatids by cleaving cohesin Rad21/SCC1/MCD1 during the metaphase to anaphase transition. The hypothesis is that hormonal stimulation of p53 null mouse mammary glands results in misexpression of the ESPL1 gene, thus promoting aneuploidy and breast cancer formation. This proposal applies in vivo transplantaion of p53 mutant and wild type (WT)mammary cells that are stably transfeeted with ESPL1, and an ESPL1 transgenic mice model to test the PRAN paradigm following hormone treatment. Steroid and p53 regulation of ESPL1 at the transcriptional level is studied by characterizing the ESPL1 promoter region. These objectives will be accomplished by pursuing two specific aims: 1) Functional role of Separaseoverexpressionin aneuploidy, and 2) Transcriptional regulation of ESPL1 gene expression. The proposed study not only elucidate underlying mechanisms of hormone-induced aneuploidy, a fundamental unresolved question in cancer biology, but also likely to identify a new class of proteins that are responsible for chromosomal instability and breast cancer progression.

该提案重点关注一个重要但尚未探索的问题——类固醇激素，它是一种很好的药物。已知的危险因素，与 p53 突变相互作用产生非整倍体和恶性肿瘤，以及如何染色体分离蛋白Separase参与其中？我们的性类固醇依赖性 p53 小鼠肿瘤前期乳腺癌模型提供了解决这个问题的独特方法。在此模型中，p53 中的类固醇诱导突变乳腺导致染色体不稳定、非整倍性和肿瘤形成，类似于大多数人类乳腺癌中都存在这种情况。我们提出了一个范式，即存在一组蛋白质其放松管制会促进非整倍性（称为 PRAN；非整倍性启动子），包括染色体不稳定性导致整个或部分染色体的丢失或增加，并且 PRAN 蛋白受到类固醇激素和肿瘤抑制因子 p53 的交互调节。我们发表的和新的初步数据提供了第一个证据，证明类固醇激素在有丝分裂蛋白的调节参与姐妹染色单体的凝聚和分离。我们建议抑癌基因 p53 突变和类固醇激素信号传导的综合作用产生通过影响染色体分离中涉及的关键蛋白的表达来抑制乳腺癌中的非整倍体。我们重点关注调节染色体分离的元素，特别是姐妹染色单体凝聚/分离蛋白，作为候选 PRAN 蛋白，因为染色体错误分离有丝分裂可导致非整倍体。该分析中的一个关键基因是 ESPL1，它编码内肽酶称为分离酶（Separase），通过在裂解过程中裂解粘连蛋白 Rad21/SCC1/MCD1 来分离连接的姐妹染色单体。中期到后期的转变。假设 p53 缺失小鼠的激素刺激乳腺导致 ESPL1 基因错误表达，从而促进非整倍体和乳腺癌形成。该提案适用于 p53 突变体和野生型 (WT) 乳腺细胞的体内移植，稳定转染 ESPL1，并建立 ESPL1 转基因小鼠模型来测试 PRAN 范式激素治疗后。研究了 ESPL1 在转录水平上的类固醇和 p53 调节表征 ESPL1 启动子区域。这些目标将通过追求两个具体目标来实现目标：1) Separase 过表达在非整倍性中的功能作用，2) ESPL1 的转录调控基因表达。拟议的研究不仅阐明了激素诱导的潜在机制非整倍性是癌症生物学中一个尚未解决的基本问题，但也可能确定一类新的负责染色体不稳定和乳腺癌进展的蛋白质。