Analysis of Intratumoral Crosstalk in Clonal Populations of OvarianTumor Cells

卵巢肿瘤细胞克隆群的瘤内串扰分析

• 批准号: 9025763
• 负责人: Joan Siefert Brugge
• 金额: $ 39.65万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-17 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9025763
• 关键词: Accounting; Affect; Ascites; Bar Codes; Biological; Blood; Bone Marrow; Brain; Cancer Control; Cells; Collaborations; Complementary DNA; Dependency; Diagnosis; Distant; Distant Metastasis; Epitopes; Genetic; Greater sac of peritoneum; Growth; Health; Heterogeneity; Histology; Human; Hypoxia; In Vitro; Individual; Liquid substance; Location; Luciferases; Lymph; Malignant Epithelial Cell; Metastatic Neoplasm to the Lung; Methodology; NOD/SCID mouse; Nature; Neoplasm Metastasis; Organ; Outcome; Ovarian Carcinoma; Peritoneal; Phenotype; Pilot Projects; Population; Primary Neoplasm; Process; Site; Solid Neoplasm; Stromal Cells; Subgroup; Testing; Time; Transplantation; Tumor Expansion; Variant; base; cancer cell; cancer therapy; insight; neoplastic cell; outcome forecast; ovarian neoplasm; tumor; tumor initiation; tumor progression; tumor xenograft

DESCRIPTION (provided by applicant): One of the most significant challenges in controlling cancer is the high degree of heterogeneity within the neoplastic cells of individual tumors. While tumor cell heterogeneity has been described at many levels and an understanding of the processes that create heterogeneity is emerging, the extent to which the heterogeneous subpopulations of tumor cells vary in their functional activities and whether interactions between subpopulation influence tumor initiation and progression is poorly understood. Major barriers to investigating these questions have been the lack of methodologies to maintain the heterogeneity of human tumors in culture and to propagate distinct clonal subpopulations from individual tumors. Together with our collaborators, we have developed methodologies that overcome these barriers and make it feasible to isolate, culture, and characterize clonal populations of ovarian tumor cells and then track barcoded, epitope-tagged clonal populations within tumor xenografts generated from mixtures of transplanted clones. Using these approaches in a pilot study, we obtained evidence for interclonal collaborations that significantly affect tumor progression - most interestingly, we found that clonal populations of cancer cells which are unable to initiate tumor formation on their own, can strongly promote the expansion of primary tumors and metastasis at specific organ sites when mixed with 'tumor-initiating' clones. This supports the hypothesis that clonal subpopulations within a tumor cooperate with one another to promote tumor expansion and metastasis. In this proposal, we describe plans to use a systematic approach to test this hypothesis in human ovarian tumors by (1) isolating and characterizing the extent of genetic and phenotypic variation among a large set of clonal populations derived from ovarian tumors, (2) comparing the activities of single clonal populations to mixtures of such populations in order to investigate the existence and nature of intratumoral cross talk, (3) tracking intratumoral localization of clonal populations over time, an (4) elucidating the mechanisms responsible for phenotypes generated by intratumoral crosstalk.

描述（由申请人提供）：控制癌症的最重大挑战之一是单个肿瘤的肿瘤细胞内的高度异质性。尽管已经在许多层面上描述了肿瘤细胞的异质性，并且对产生异质性的过程的理解正在出现，但肿瘤细胞的异质亚群在其功能活性中的异质亚群在其功能活性中的不同程度以及亚群之间的相互作用是否影响肿瘤起始和进展是否不足。研究这些问题的主要障碍是缺乏维持培养物中人类肿瘤异质性并传播各个肿瘤不同克隆亚群的方法论。与我们的合作者一起，我们开发了克服这些障碍的方法，并使卵巢肿瘤细胞的克隆人群隔离，培养和表征可行，然后跟踪从移植式锁骨的混合物中产生的肿瘤异种移植物中的条形码，表位，表位标记的克隆种群。在一项试点研究中，我们获得了这些方法，我们获得了克隆间协作的证据 影响肿瘤的进展 - 最有趣的是，我们发现无法自行启动肿瘤形成的癌细胞的克隆种群可以强烈促进特定器官的原发性肿瘤和转移的扩张，当时与“肿瘤炎性”克隆混合时。这支持了以下假设：肿瘤内的克隆亚群相互配合以促进肿瘤的扩张和转移。在该提议中，我们描述了使用系统的方法来检验人类卵巢肿瘤中这种假设的计划，通过（1）隔离和表征遗传和表型变化的程度，在卵巢肿瘤中得出的大量克隆人群中的遗传和表型变化的程度，（2）将单个间接种群的活性与本内部的群体进行比较（2），并将其内部构成（3）进行比较。克隆人群的定位随着时间的流逝，（4）阐明了负责由肿瘤内串扰产生的表型的机制。