Investigating the roles of oncogenic extrachromosomal circular DNAs in cancer

研究致癌染色体外环状 DNA 在癌症中的作用

基本信息

批准号：
10718423
负责人：
Andrea Ventura
金额：
$ 56.67万
依托单位：
SLOAN-KETTERING INST CAN RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2028-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10718423
关键词：
Acceleration Address Advanced Malignant Neoplasm Animals Appearance Applications Grants Biogenesis Biology Cancer cell line Cell Line Cells Chromosomal Duplication Chromosomal Rearrangement Chromosomes Circular DNA Clustered Regularly Interspaced Short Palindromic Repeats Cues DNA Damage DNA Maintenance Data Double Minutes Drug resistance Elements Engineering Environment Epidermal Growth Factor Receptor Epigenetic Process Event Evolution Future Generations Genes Genetic Genetic Recombination Genetically Engineered Mouse Genome engineering Goals Horizontal Disease Transmission Human Investigation Label Lesion Link MDM2 gene Maintenance Malignant Neoplasms Mediating Metaphase Spread Mitosis Modeling Mouse Strains Mus Neoplasm Metastasis Normal Cell Nutrient Oncogene Activation Oncogenes Oncogenic Other Genetics Pathogenesis Phenotype Reagent Reporter Research Proposals Role Series System Technology Testing Therapeutic Tumor Promotion Tumor stage Visualization Work anti-cancer therapeutic cancer cell cell transformation daughter cell erbB Genes experimental study extracellular extrachromosomal DNA genetic approach genetic element genome editing human cancer mouse model in vivo inducible Cre innovation interest metaplastic cell transformation model organism novel novel strategies prevent segregation somatic cell gene editing tool tumor tumor heterogeneity tumor initiation tumor progression tumorigenesis

项目摘要

ABSTRACT Increased oncogene expression mediated by focal amplifications is a common mechanism for oncogene activation in human cancers. Two major mechanisms leading to oncogene amplification have been described: chromosomal amplification and non-chromosomal amplification. The latter mechanism is characterized by the presence of multiple copies of circular DNAs that are thought to originate following the fragmentation and subsequent circularization of pieces of chromosomes. These “extrachromosomal circular DNAs” (ecDNAs) have long been known as “double minutes” for their appearance in metaphase spreads and by the lack of centromeric sequences. In the past few years renewed interest in this class of cancer-associated chromosomal rearrangements has been fueled by technological advances and by the realization that, due to their random segregation at mitosis, ecDNAs can accelerate tumor evolution, mediate drug resistance, and generally promote a more aggressive phenotype. Despite substantial progress, however, several key questions regarding the biology of ecDNAs, their dynamics during the early stages of tumor formation, and their contribution to tumor initiation and progression, remain unanswered. This is in part due to the lack of effective means to engineer and track ecDNAs in normal cells and in model organisms. Our group has extensive expertise in the generation and characterization of germline and somatic mouse models of human cancers, and we have pioneered the use of somatic genome editing to engineer chromosomal rearrangements in mice. In this grant application, supported by strong preliminary data, we describe a novel general strategy to model ecDNAs in cells and in mice. We have already generated three new genetically engineered mouse strains in which the formation of ecDNAs containing the oncogenes most commonly amplified in human cancers can be induced in a temporally and spatially controlled manner. Using a similar strategy, we have also generated cell lines in which formation of specific ecDNAs can be induced and tracked non-invasively using fluorescent reporters and selectable markers. We propose to use these innovative tools and reagents to address the following key questions: 1) Can oncogenic ecDNAs initiate tumor formation and/or accelerate tumor progression and metastasis in vivo? 2) How do oncogenic ecDNAs respond to changes in intracellular and extracellular environment? 3) Are there mechanisms preventing ecDNA formation and propagation in primary cells? 4) Is the presence of ecDNAs in cancer cells associated with unique therapeutically actionable vulnerabilities? 5) Can ecDNAs be transmitted horizontally between cells?

抽象的局灶扩增介导的癌基因表达增加是癌基因的常见机制已经描述了导致癌基因扩增的两种主要机制：染色体扩增和非染色体扩增的特点是。存在多个环状 DNA 拷贝，这些拷贝被认为起源于片段化和这些“染色体外环状DNA”（ecDNA）随后发生环化。长期以来被称为“双分钟”，因为它们出现在中期扩散中并且缺乏着丝粒序列。在过去的几年里，人们对这类与癌症相关的染色体重排重新产生了兴趣。在技术进步的推动下，以及认识到，由于 ecDNA 在有丝分裂时随机分离，可以加速肿瘤进化，介导耐药性，并通常促进更具侵袭性的表型。尽管取得了实质性进展，然而，关于 ecDNA 生物学、其动力学的几个关键问题在肿瘤形成的早期阶段，它们对肿瘤发生和进展的贡献仍然存在这部分是由于缺乏有效的方法来设计和追踪正常细胞中的 ecDNA。在模式生物中。我们的团队在种系和体细胞小鼠模型的生成和表征方面拥有丰富的专业知识人类癌症，我们率先使用体细胞基因组编辑来改造染色体在这项拨款申请中，在强有力的初步数据的支持下，我们描述了一种新颖的重组方法。在细胞和小鼠中模拟 ecDNA 的一般策略。我们已经培育出三种新的基因工程小鼠品系，其中 ecDNA 的形成含有在人类癌症中最常扩增的癌基因可以在暂时和使用类似的策略，我们还生成了其中形成的细胞系。可以使用荧光产生器和选择标记非侵入性地诱导和追踪特定的 ecDNA。我们建议使用这些创新工具和试剂来解决以下关键问题： 1) 致癌 ecDNA 能否在体内启动肿瘤形成和/或加速肿瘤进展和转移？ 2）致癌ecDNA如何响应细胞内和细胞外环境的变化？ 3）是否存在阻止原代细胞中ecDNA形成和增殖的机制？ 4) 癌细胞中 ecDNA 的存在是否与独特的治疗可行的脆弱性相关？ 5）ecDNA可以在细胞之间水平传输吗？