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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要癌症的特点是基因组、表观遗传和代谢变化，但也包括细胞/核大小的变化控制。然而，人们对癌细胞的大小调节以及大小控制是否失败知之甚少。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的中间事件是可以改变细胞和细胞核大小的关键过程。然而，虽然在正常人类细胞中，倍性和核大小往往成比例缩放，但 DNA 含量和核大小则不然总是与癌细胞相关。这表明了解核大小是如何控制的 DNA含量的变化可能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，例如将用于该项目的模型。该模型包括通过在结直肠癌和乳腺癌中诱导 WGD 衍生的一组四倍体 (4N) 克隆细胞系细胞可以根据核大小分为两组（小组和大组）。先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，这提出了重要的问题 4N 细胞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，小型 4N 克隆在体外具有增强的肿瘤样行为，表现优于软琼脂集落形成测定中的大 4N 克隆和 2N 亲代细胞。这些特定大小的表型差异表明，小 4N 克隆在体内也可能比大 4N 克隆更具致瘤性，因为归功于他们在 WGD 后限制核尺寸缩放的能力。该假设将在第一个研究目标中得到检验结合使用体外、体内和计算机方法以及两种实验模型，包括新颖的源自 2N DLD1 细胞（总共 16 个细胞系）和基因组的小型和大型 4N 克隆组，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 克隆的核体积由基因组压缩程度决定，并且将确定结肠和乳腺癌全基因组测序后调节核大小的转录和表观遗传机制衍生细胞。这些实验将解决一个基本但尚未充分探索的问题：变异是否核大小的变化具有重要的功能后果，有助于 WGD 后肿瘤的形成。这该项目的研究结果可用于癌症诊断并揭示新的癌症治疗靶点。这项目将提供充足的动物模型使用、计算分析和质量方面的培训机会由领先研究人员组成的跨学科小组进行光谱测定，以帮助申请人成为独立的科学家。为申请者提供的智力环境、资源和培训项目丰富、对拟议的研究项目的补充。此外，集体制度环境强烈支持多学科研究，有助于拟议研究的成功和申请人的科学发展。