An in vivo model for CCNE1 amplified tumorigenesis

CCNE1 放大肿瘤发生的体内模型

基本信息

批准号：
9396699
负责人：
Paul T Kroeger
金额：
$ 6.1万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-12-01 至 2020-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9396699
关键词：
Adjuvant Chemotherapy Alleles Animal Model Animals BRCA1 gene BRCA2 gene Biological Assay CCNE1 Gene Amplification CCNE1 gene Carcinoma in Situ Cell Cycle Cell division Cells Cessation of life Complementary DNA Cytotoxic Chemotherapy DNA Repair Pathway Data Databases Development Diagnosis Disease Doxycycline Engineering Enterobacteria phage P1 Cre recombinase Epithelial Cells Epithelium Event Fimbriated End of the Fallopian Tube Generations Genes Genetic Screening Genetically Engineered Mouse Genome Genomics Grant Histologic Human In Vitro International Knock-out Laboratories Laboratory Study Length Lesion Malignant Female Reproductive System Neoplasm Malignant Neoplasms Malignant neoplasm of ovary Mammalian Oviducts Mediating Methodology Modeling Molecular Genetics Molecular Target Molecular Weight Mus Mutation Neoplastic Cell Transformation Outcome Ovarian Ovarian Serous Adenocarcinoma Ovary Pathogenesis Patients Pharmaceutical Preparations Platinum Poly(ADP-ribose) Polymerases Proteins Proteomics Public Health Recurrent disease Reproducibility Research Design Resistance Resources Risk Schools Secretory Cell Serous Solid Surface TP53 gene Technology Testing The Cancer Genome Atlas Tissues Transgenic Mice Transgenic Model Triage Tube Tumor Debulking Tumor Suppressor Genes United States Woman brca gene cancer genome cell injury chemotherapy effective therapy experience experimental study homologous recombination human disease in vivo Model inhibitor/antagonist insight loss of function malignant breast neoplasm model development mortality mouse model mutant novel novel therapeutics ovarian neoplasm overexpression patient population pre-clinical progenitor promoter protein protein interaction recombinational repair response standard care therapeutic target tool tumor tumorigenesis

项目摘要

PROJECT SUMMARY High-grade serous ovarian carcinoma (HGSOC) is one of the most devastating cancer-related diseases in the United States. It is the deadliest gynecological malignancy and a public health burden. Historically, it was thought that ovarian tumors arise from the ovarian surface epithelium. Recently, this school of thought has been challenged by the finding of early HGSOC precursors in the fallopian tubes (FT) of women at increased risk of developing HGSOC due to mutations in the BRCA1 or BRCA2 genes. Most cases were localized to the fimbriated end of the FT and included serous tubal intraepithelial carcinoma (STIC) as the dominant precursor lesion. The Cancer Genome Atlas of HGSOC showed that approximately 50% of HGSOCs harbor mutations in the BRCA genes or other genes in the homologous recombination (HR) pathway of DNA repair. These tumors tend to respond well to chemotherapy and poly (ADP-ribose) polymerase (PARP) inhibitors. Unfortunately, the remaining 50% of HGSOC are often HR-proficient, do not respond well to standard treatment, and are associated with worse outcomes. HGSOCs with amplification of the CCNE1 gene represent a significant fraction of these HR-proficient tumors. CCNE1 makes a protein, cyclinE1, that controls cell division but which can also damage the cell's genome when present in excess. Women with CCNE1 amplified tumors are unlikely to respond to PARP inhibitors and thus represent an important unmet need. While there are a number of drugs currently in development in other solid cancers that may be effective in CCNE1 amplified HGSOC, there are currently no animal models to study the function of cyclin E1 in HGSOC or the efficacy of candidate inhibitors. This application describes our aim to develop a mouse model that mimics the human disease, thereby providing a preclinical platform for testing novel cyclinE1 inhibitors. We will construct transgenic mice where the Ccne1 gene can be precisely activated when and where we want it. Using well-established technology, we will engineer mice to express the cyclinE1 protein at high levels in FT secretory epithelial cells, the progenitors of HGSOC. We are very experienced in the generation of such animals and their analysis. Importantly, TP53 is always defective in human HGSC and its loss appears to be a requirement for cells to tolerate the presence of excess cyclinE1. Therefore, our model will incorporate expression of a mutant Tp53. Our laboratory studies suggest these two genetic alterations – Ccne1 over expression and Tp53 mutation – will be enough to generate tumors. It is also possible, however, that additional mutations will be needed for the mice to develop HGSC that mimic the human disease. In the second part of the grant we use data from a genetic screen in FT cells and information from thousands of cancer genomes to find genes that may cooperate with CCNE1 and that could be added to the mice. The study is likely to provide a high return on the invested effort, as the mice will provide an enduring resource - once the model is established it can be used in many settings, providing a powerful ongoing platform for development of CCNE1 inhibitors.

项目摘要高级浆液卵巢癌（HGSOC）是与癌症有关的疾病之一美国。这是最致命的妇科恶性肿瘤和公共卫生伯恩。从历史上看，是认为卵巢肿瘤来自卵巢表面上皮。最近，这个思想流派有在增加的女性输卵管（FT）中发现了早期的HGSOC前体的挑战由于BRCA1或BRCA2基因突变引起的HGSOC的风险。大多数案例都本地化 FT的纤维化端，包括浆液管内癌（Stic）作为主要前体病变。 HGSOC的癌症基因组图谱显示，大约50％的HGSOC携带突变 DNA修复的同源重组（HR）途径中的BRCA基因或其他基因。这些肿瘤倾向于对化学疗法和聚（ADP-核糖）聚合酶（PARP）抑制剂的反应良好。不幸的是，剩下的50％的HGSOC通常是HR的，对标准治疗的反应不好，并且是与较差的结果相关。 CCNE1基因扩增的HGSOC表示显着这些HR肿瘤的一部分。 CCNE1生成蛋白质Cycline1，可控制细胞分裂，但当过量存在时，还会损害细胞的基因组。 CCNE1扩增肿瘤的女性不太可能响应PARP抑制剂，因此代表了重要的未满足需求。虽然有很多药物目前正在其他固体癌症中正在开发，这些癌症可能有效地在CCNE1扩增HGSOC中目前，尚无动物模型来研究细胞周期蛋白E1在HGSOC中的功能或候选抑制剂的效率。该应用描述了我们开发模仿人类疾病的小鼠模型的目的提供一个用于测试新型Cycline1抑制剂的临床前平台。我们将在其中建造转基因小鼠 CCNE1基因可以在我们想要的何时何地被精确激活。使用公认的技术，我们会在FT分泌上皮细胞（祖细胞）中高水平以高水平表达Cycline1蛋白 HGSOC。我们在这样的动物及其分析方面非常有经验。重要的是，TP53是始终在人类HGSC中有缺陷，其损失似乎是细胞忍受存在的必要条件多余的环素1。因此，我们的模型将结合突变体TP53的表达。我们的实验室研究建议这两个遗传改变 - CCNE1超过表达和TP53突变 - 足以产生肿瘤。但是，也可能需要其他突变才能开发出来模仿人类疾病的HGSC。在赠款的第二部分中，我们使用FT中遗传屏幕的数据细胞和来自数千种癌症基因组的信息，以查找可能与CCNE1和CCNE1协调的基因可以添加到老鼠中。这项研究可能会为投资努力提供高回报，因为老鼠将提供持久的资源 - 建立模型后，它可以在许多设置中使用，提供强大的持续开发CCNE1抑制剂的平台。