In vivo imaging of tumor heterogeneity generation from endogenous single cells

内源性单细胞肿瘤异质性产生的体内成像

• 批准号: 9178032
• 负责人: SCOTT E FRASER
• 金额: $ 21.53万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9178032
• 关键词: Address; Adult; Animals; Automobile Driving; Back; Benign; Biological; Blood Vessels; Categories; Cell Differentiation process; Cell Lineage; Cell Proliferation; Cell division; Cells; Characteristics; Detection; Development; Developmental Biology; Distal; Drug resistance; Epidermis; Epithelium; Flow Cytometry; Freezing; Gene Expression; Generations; Genes; Heterogeneity; Image; Image Analysis; Individual; Invaded; Investigation; Label; Lead; Life; Link; Lymph; Malignant - descriptor; Malignant Neoplasms; Methods; Modeling; Molecular; Neoplasm Metastasis; Oncogenes; Pathway Analysis; Pathway interactions; Phenotype; Population; Process; Proliferating; Property; Proteins; Reporter; Role; Sequence Analysis; Skin; Stem cells; System; Therapeutic; Tissues; Transgenic Organisms; Tumor Angiogenesis; Tumor Cell Invasion; Tumor-Associated Process; Tumor-Derived; Work; Zebrafish; anti-cancer therapeutic; base; benign state; cancer imaging; cancer stem cell; cell behavior; cell type; early onset; in vivo; in vivo imaging; interest; mature animal; molecular marker; neoplastic cell; next generation sequencing; non-invasive imaging; novel; prevent; research study; self-renewal; tool; transcriptome; tumor; tumor growth; tumor heterogeneity; tumor microenvironment; tumorigenesis

PROJECT SUMMARY The formation of a malignant tumor from a benign tumor has been strongly linked to the existence of heterogeneous cellular features within the tumor. Thus, elucidating the mechanisms underlying generation of tumor cell heterogeneity will be an important milestone to understand malignant tumor formation. We believe two applications of our work could be 1) earlier, more quantitative detection of malignancy based on molecular markers and 2) therapeutics that can freeze tumor development before metastasis or even revert back to a benign state. Our approach is to answer the following two questions: 1. What is the impact of the tumor-initiating cell‘s identity on generating tumor cell heterogeneity? In experiments to date, the inability to avoid contamination of different cell origins, including tissue stem cells and non-stem cells into the resultant tumor mass has prevented identification of the precise molecular mechanisms. We address the contamination problem by establishing an experimental system that enables precise tracing of tumor cell lineage derived from single cells. Our method is the non-invasive, in vivo imaging of endogenous, single cell-derived tumor development in adult transparent zebrafish. This experimental system allows us to visualize individual tumor cells, and, more importantly, also selectively label and isolate the cells of interest for detailed cell characterization. Using this system, we have already imaged the tumors arising from single cells in the skin epithelium. Importantly, we have found some major differences in tumor cell proliferation between tumorous cell clusters and in tumor cell differentiation that lead to acquisition of invasive features. Our results identifying a subset of tumor cells displaying invasive features is nicely representing the generation of tumor cell heterogeneity within a tumor. We will perform long-term clonal analysis of tumor development and transcriptome analysis of isolated individual tumors to determine their cellular identity. Retrospective imaging analysis will be used to study the effect of a tumor’s microenvironment on generating tumor cell heterogeneity. 2. How is heterogeneity within a tumor established? Identification of the cell types of tumor-initiating cells will be followed by the investigation of the mechanisms driving a single tumor-initiating cell to become heterogeneous during multiple rounds of cell divisions. Our ability to distinctively label and isolate invasive tumor cells from non-invasive tumor cells enables us to investigate the gene expression modulations responsible for a subset of tumor cells acquiring invasive features during tumor development. We will further perform functional experiments in vivo to define the roles of specific molecular pathways in tumor invasion.

项目摘要 来自良性肿瘤的恶性肿瘤形成与存在密切相关 肿瘤内的异质细胞特征。那，阐明了生成的机制 肿瘤细胞异质性将是了解恶性肿瘤形成的重要里程碑。我们相信 我们工作的两个应用可能是1）早期，基于分子的更定量检测恶性肿瘤 标记和2）可以在转移之前冻结肿瘤发展的疗法，甚至还原回 良性国家。我们的方法是回答以下两个问题： 1。肿瘤发射细胞的身份对产生肿瘤细胞异质性有什么影响？ 在迄今为止的实验中，无法避免污染不同的细胞起源，包括组织干细胞 和非茎细胞进入所得的肿瘤质量已阻止了精确分子的鉴定 机制。我们通过建立一个实验系统来解决污染问题 源自单细胞的肿瘤细胞谱系的精确追踪。我们的方法是无创的，体内成像的 成人透明斑马鱼的内源性，单细胞衍生的肿瘤发育。这个实验系统 允许我们可视化单个肿瘤细胞，更重要的是，也有选择地标记和分离 详细的细胞表征的兴趣。使用此系统，我们已经对肿瘤成像 皮肤上皮中的单细胞。重要的是，我们发现肿瘤细胞增殖有一些主要差异 在肿瘤细胞簇和肿瘤细胞分化之间导致侵入性特征的获取。我们的 识别显示出侵入性特征的肿瘤细胞子集的结果很好地表示 肿瘤中的肿瘤细胞异质性。我们将对肿瘤发育的长期克隆分析和 分离的个体肿瘤的转录组分析以确定其细胞身份。追溯成像 分析将用于研究肿瘤微环境对产生肿瘤细胞异质性的影响。 2。在肿瘤内如何建立异质性？ 鉴定肿瘤发射细胞的细胞类型，将研究机理 在多轮细胞分裂期间，驱动单个肿瘤发射细胞成为异质。我们的能力 从非侵入性肿瘤细胞中明显标记和分离出浸润性肿瘤细胞，使我们能够研究 负责肿瘤细胞子集的基因表达调制，使肿瘤中侵入性特征加速 发展。我们将进一步在体内执行功能实验，以定义特定分子的作用 肿瘤入侵的途径。