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

• 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
• 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

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 通过局灶性扩增介导的癌基因表达增加是癌基因的常见机制 在人类癌症中激活。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。后一种机制的特征是 存在多个圆形DNA的副本，这些圆形DNA被认为是在碎片化和 随后的染色体循环。这些“外圆形圆形DNA”（ECDNA）具有 长期以来被称为“双分钟”，因为它们出现在中期扩散和缺乏centromeric中 序列。 在过去的几年中，对这类癌症相关的染色体重排的兴趣已 在技​​术进步的推动下，并意识到，由于它们在有丝分裂时的随机分离，ECDNA 可以加速肿瘤的演化，介导耐药性，并通常促进更具侵略性的表型。 尽管取得了长足的进步，但关于ECDNA的生物学的几个关键问题，他们的动态 在肿瘤形成的早期及其对肿瘤倡议和进展的贡献中，仍然存在 未得到答复。这部分是由于缺乏有效的手段来设计和跟踪正常细胞中的ECDNA和 在模型生物中。 我们的小组在生殖线和体细胞模型的产生和表征方面具有广泛的专业知识 人类的癌症，我们已经开创了将体细胞基因组编辑用于染色体的使用 小鼠重排。在此赠款应用程序中，在强大的初步数据的支持下，我们描述了一个小说 在细胞和小鼠中模拟ECDNA的一般策略。 我们已经产生了三种新的一般工程的鼠标菌株，其中ecdnas的形成 可以在临时和 空间控制的方式。使用类似策略，我们还生成了细胞系 可以使用荧光记者和可选标记来诱导特定的ECDNA并非诱导和跟踪。 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）致癌性ECDNA可以启动肿瘤形成和/或体内加速肿瘤进展和转移吗？ 2）致癌性ECDNA如何应对细胞内和细胞外环境的变化？ 3）是否有防止原代细胞中ecDNA形成和传播的机制？ 4）在癌细胞中存在ECDNA是否与独特的治疗性可行的脆弱性相关？ 5）可以在细胞之间水平传播ECDNA吗？