Understanding how variations in nuclear size after whole genome doubling affect tumorigenesis

了解全基因组加倍后核大小的变化如何影响肿瘤发生

• 批准号: 10607178
• 负责人: Mathew Bloomfield
• 金额: $ 4.19万
• 依托单位: VIRGINIA POLYTECHNIC INST AND ST UNIV
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-10 至 2025-04-09
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10607178
• 关键词: Address; Affect; Agar; Anchorage-Independent Growth; Aneuploidy; Animal Model; Animals; Bioinformatics; Biological Assay; Biopsy; Breast; Breast Cancer Cell; Breast Cancer cell line; Breast Epithelial Cells; Cancer Diagnostics; Cancer cell line; Cell Culture Techniques; Cell Line; Cell Nucleus; Cell Size; Cells; Cessation of life; Chemicals; Clinical; Clinical Data; Clone Cells; Colon; Colon Carcinoma; Colorectal Cancer; Computer Analysis; Cytokinesis; Cytoplasm; DNA Methylation; Data; Development; Diploidy; Disease; Environment; Epigenetic Process; Event; Experimental Models; Failure; Gene Expression; Gene Expression Profiling; Genes; Genetic; Genetic Transcription; Genome; Genomics; HCT116 Cells; Histones; Histopathology; Human; Image; In Vitro; Institution; Interdisciplinary Study; Invaded; Link; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Metabolic; Metabolism; Metastatic breast cancer; Methods; Mitotic; Modeling; Mus; Nature; Nuclear; Phenotype; Ploidies; Population; Post-Translational Protein Processing; Postdoctoral Fellow; Prevalence; Process; Regulation; Research; Research Personnel; Research Project Grants; Resources; Running; Scientist; Shapes; Soft tissue sarcoma; Technical Expertise; Testing; Tetraploidy; The Cancer Genome Atlas; Therapeutic; Training; Training Programs; Tumorigenicity; United States; Variant; Work; Xenograft procedure; behavior in vitro; cancer cell; career; cell type; digital; epigenomics; experimental study; fitness; genomic data; in silico; in vivo; insight; interest; neoplastic cell; novel; success; therapeutic target; training opportunity; tumor; tumor progression; tumor xenograft; tumorigenesis; tumorigenic; whole genome

Project Summary Cancer is characterized by genomic, epigenetic, and metabolic changes, but also by a failure in cell/nuclear size control. Nevertheless, little is known about size regulation in cancer cells and whether failures in size control contribute to tumorigenesis. Whole genome doubling (WGD), which results in tetraploidy and is a frequent intermediate event in tumor progression, is a key process that can alter cell and nuclear size. However, while ploidy and nuclear size tend to scale proportionally in normal human cells, DNA content and nuclear size do not always correlate in cancer cells. This suggests that understanding how nuclear size is controlled following changes in DNA content may uncover important mechanisms underlying tumorigenesis. Addressing this issue requires a suitable experimental model, such as the one that will be used for this project. This model consists of a panel of tetraploid (4N) clonal cell lines that were derived by inducing WGD in colorectal and breast cancer cells and can be divided into two groups (small and large) based on nuclear size. Previous studies showed that the small and large 4N DLD1 clones display distinct mitotic phenotypes, which raised important questions about the mechanisms of size control in 4N cells and the impact of nuclear size on other cell phenotypes. Indeed, preliminary results indicate that the small 4N clones have increased tumor-like behavior in vitro, outperforming the large 4N clones and 2N parental cells in soft agar colony formation assays. These size-specific phenotypic differences suggest that the small 4N clones may also be more tumorigenic in vivo than the large 4N clones due to their ability to restrict nuclear size scaling after WGD. This hypothesis will be tested in a first research aim using a combination of in vitro, in vivo, and in silico methods and two experimental models, including the novel panel of small and large 4N clones derived from 2N DLD1 cells (for a running total of 16 cell lines) and genomic, histopathology, and clinical data from The Cancer Genome Atlas (TCGA). A second research aim will test the hypothesis that the nuclear volumes of the 4N clones are dictated by the degree of genomic compaction and will identify the transcriptional and epigenetic mechanisms that regulate nuclear size after WGD in colon- and breast- derived cells. These experiments will address a fundamental yet underexplored question of whether variations in nuclear size have important functional consequences that contribute to tumor formation following WGD. The findings of this project could be leveraged for cancer diagnostics and unveil new cancer therapeutic targets. This project will provide ample training opportunities in the use of animal models, computational analysis, and mass spectrometry from an interdisciplinary group of leading researchers to help the applicant become an independent scientist. The intellectual environment, resources, and training programs provided to the applicant are rich and complementary to the proposed research project. Moreover, the collective institutional environment is strongly supportive of multidisciplinary research, and it is instrumental to the success of the proposed studies and scientific development of the applicant.

项目摘要 癌症的特征是基因组、表观遗传学和新陈代谢的改变，但也有细胞/核大小的缺陷。 控制力。然而，对于癌细胞的大小调节以及大小控制的失败，人们知之甚少。 有助于肿瘤的发生。全基因组加倍(WGD)，导致四倍体，是一种常见的 肿瘤进展中的中间事件，是改变细胞和核大小的关键过程。然而，虽然 在正常人类细胞中，倍性和核大小倾向于成比例地扩大，而DNA含量和核大小则不是 在癌细胞中总是相关的。这表明，理解核大小是如何控制的 DNA含量的变化可能会揭示肿瘤发生的重要机制。解决这个问题 需要一个合适的实验模型，例如将用于此项目的模型。该模型由以下部分组成 一组通过诱导结直肠癌和乳腺癌WGD而获得的四倍体(4N)克隆细胞系 根据核的大小，可以将其分为两组(小的和大的)。此前的研究表明， 小的和大的4N DLD1克隆表现出不同的有丝分裂表型，这提出了关于 4N细胞的大小控制机制以及核大小对其他细胞表型的影响。的确， 初步结果表明，小的4N克隆在体外增加了肿瘤样行为，表现优于 软琼脂集落形成实验中获得大的4N克隆和2N亲本细胞。这些尺码特定的表型 差异表明，小的4N克隆在体内也可能比大的4N克隆在体内更具致瘤性 他们有能力在WGD之后限制核尺寸的扩大。这一假设将在第一个研究目标中得到检验。 使用体外、体内和硅胶方法和两个实验模型相结合的方法，包括新的 来自2N个DLD1细胞(总共16个细胞系)和基因组的4N个小克隆和大克隆的小组， 组织病理学和来自癌症基因组图谱(TCGA)的临床数据。第二个研究目标将测试 假设4N个克隆的核体积由基因组压缩的程度和意志决定 确定WGD后结肠和乳房中调节核大小的转录和表观遗传机制 派生的细胞。这些实验将解决一个基本但未被探索的问题，即变异 核的大小对WGD后肿瘤的形成有重要的功能影响。这个 该项目的发现可能被用于癌症诊断，并揭示新的癌症治疗目标。这 该项目将提供大量使用动物模型、计算分析和质量的培训机会 来自一个由领先研究人员组成的跨学科小组，帮助申请者成为一名独立的 科学家。为申请人提供的智力环境、资源和培训方案丰富， 这是对拟议研究项目的补充。此外，集体制度环境强烈。 支持多学科研究，这有助于拟议研究的成功和 申请人的科学发展。