A Cellular and Molecular Analysis of the Intravasation Step in Tumor Metastasis

肿瘤转移中浸润步骤的细胞和分子分析

• 批准号: 7915828
• 负责人: JAMES P QUIGLEY
• 金额: $ 34.47万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7915828
• 关键词: Animals; Biological Assay; Blood; Blood Vessels; Carcinoma; Cell Communication; Cell Line; Cell physiology; Cells; Cessation of life; Chick Embryo; Clinical; Complement; Databases; Distal; Down-Regulation; Embryo; Evaluation; Event; Exhibits; Extravasation; Fibroblast Growth Factor 2; Fibroblasts; Gelatinase B; Goals; Human; In Vitro; Individual; Inflammatory; Intervention; Knowledge; Laboratories; Link; Malignant Epithelial Cell; Malignant Neoplasms; Matrix Metalloproteinases; Measurement; Mesenchymal; Modeling; Modification; Molecular; Molecular Analysis; Monitor; Mus; Neoplasm Metastasis; Organ; Outcome; Pancreatic carcinoma; Pathologic Processes; Physiological; Population; Primary Neoplasm; Procedures; Process; Property; Prostate; Proteins; Proteomics; Reporting; Research; Role; Site; Stromal Invasion; Survival Rate; Testing; Tissues; Tumor Cell Invasion; Variant; angiogenesis; cancer type; chorioallantoic membrane; comparative; congenic; fibrosarcoma; in vivo; insight; intravital microscopy; mouse model; neoplastic cell; novel; public health relevance; repository; tumor

DESCRIPTION (provided by applicant): A number of the specific steps in tumor dissemination have been extensively modeled, and molecularly dissected. However, one early step in the metastatic cascade, namely intravasation, the entry of escaping tumor cells into the vasculature, has been relatively understudied. One of the reasons for this is that up to now human tumor cell phenotypic variants that exhibit substantial differences in their intravasation ability have not been available for comparative analysis. Our laboratory reported the selection and isolation of two variants of a human fibrosarcoma cell line (HT-1080) that dramatically differ (50-100 fold) in their intravasation rate, yielding a similar >50 fold differential in their metastatic capabilities. These two tumor dissemination variants, HT- hi/diss and HT-lo/diss, were selected and monitored in the chick embryo model, where primary human tumors developing on the embryo's chorioallantoic membrane (CAM), recapitulate the multi-step tumor dissemination process and form micro metastatic foci in a number of secondary organs. We have also tested the two fibrosarcoma variants in different mouse metastasis assays and confirmed the substantial differential in their tumor dissemination capabilities. These two congenic, intravasation variants are thus suitable for a comparative analysis of early metastatic events. Therefore, in Specific Aim 1 we propose to molecularly dissect three physiological/pathological processes which determine the outcome of tumor cell intravasation events, i.e. induction of angiogenesis, stromal invasion and vasculotropism. We will analyze the functional role of select molecules contributing to the events involved in tumor cell intravasation including; inflammatory cell MMP-9; FGF-2; uPA; and tumor cell MMP-14. Unique assays for tumor cell interaction with blood vessels will be used to identify contributory molecules involved in the vasculotropic event. Mouse models for tumor cell dissemination and angiogenesis will be used to complement the chick embryo models for examining the identified molecules in the two fibrosarcoma variants. There exists however, a distinct lack of other tumor cell intravasation variants to compare and contrast and this is especially relevant for carcinomas, the major cancer in the human population. Therefore for Specific Aim 2 we propose to identify specific proteins which contribute mechanistically to the intravasation step in carcinoma dissemination by employing carcinoma variants selected in vivo for differential rates of intravasation. We will generate pairs of high and low intravasating variants from human prostate, colon and pancreatic carcinoma cell lines. Functional proteomic approaches will be applied both in vitro and in vivo to the selected carcinoma variants to verify and/or complement the cellular mechanisms and contributory molecules identified in fibrosarcoma-derived variants. The influence of mesenchymal fibroblasts and their products on the disseminating properties of the carcinoma variants will also be quantified. Novel mechanistic information about carcinoma cell intravasation will provide molecular links to a specific step in tumor dissemination, namely intravasation. PUBLIC HEALTH RELEVANCE: Deaths from cancer occur mainly because tumor cells spread from the primary tumor site to other vital organs and tissues. The tumor spread is usually through the vasculature. In order for tumor cells to escape from the primary tumor and enter the vasculature they alter some of their cellular processes by producing different levels of functioning molecules. The goal of the proposed research is to identify the relevant molecules and cellular processes that contribute to tumor cell spread so that clinical intervention can target those critical molecules and processes.

描述（由申请人提供）：已经对肿瘤传播中的许多特定步骤进行了广泛的建模和分子解剖。然而，在转移性级联反应的早期一步，即侵入，将肿瘤细胞逃脱到脉管系统中，已经相对研究了。这样做的原因之一是，直到现在的人类肿瘤细胞表型变体在其侵入能力上表现出很大的差异尚未用于比较分析。我们的实验室报告了人纤维肉瘤细胞系（HT-1080）的两种变体的选择和分离，它们的侵入率很大差异（50-100倍），在转移性能力中产生了相似的> 50倍差异。在雏鸡胚胎模型中选择并监测了这两个肿瘤传播变体HT-HI/DISS和HT-LO/DISS，在该模型中，在胚胎的shorioallantoicic膜（CAM）上发育的原发性人类肿瘤（CAM）概述了多步肿瘤传播过程，并形成了微型转移灶，并在次要的次要官方数量中形成了微型转移灶。我们还测试了不同小鼠转移测定法中的两个纤维肉瘤变体，并证实了其肿瘤传播能力的实质性差异。因此，这两种相互侵入的变体适合对早期转移事件的比较分析。因此，在特定目标1中，我们建议分子剖析三个确定肿瘤细胞插入事件结果的生理/病理过程，即诱导血管生成，基质侵袭和血管质主义。我们将分析有助于参与肿瘤细胞插入的事件的精选分子的功能作用，包括：炎症细胞MMP-9; FGF-2; upa;和肿瘤细胞MMP-14。肿瘤细胞与血管相互作用的独特测定将用于鉴定脉管型事件中涉及的促进分子。肿瘤细胞传播和血管生成的小鼠模型将用于补充雏鸡胚胎模型，以检查两个纤维肉瘤变体中鉴定的分子。然而，存在明显的缺乏与其他肿瘤细胞插入变体进行比较和对比，这与人群中的主要癌症癌尤其重要。因此，对于特定目标2，我们建议鉴定特定的蛋白质，这些蛋白质通过使用在体内选择的癌变体来促进癌传播的侵入性蛋白质。我们将产生来自人类前列腺，结肠和胰腺癌细胞系的高和低静脉变体。功能性蛋白质组学方法将在体外和体内应用于选定的癌变体，以验证和/或补充在纤维肉瘤衍生的变体中鉴定的细胞机制和促进分子。间充质成纤维细胞及其产物对癌变体的传播特性的影响也将进行量化。有关癌细胞侵入的新机械信息将提供与肿瘤传播的特定步骤的分子联系，即插入。公共卫生相关性：癌症死亡的原因主要是因为肿瘤细胞从原发性肿瘤部位扩散到其他重要器官和组织。肿瘤扩散通常是通过脉管系统。为了使肿瘤细胞从原发性肿瘤中逸出并进入脉管系统，它们通过产生不同水平的功能分子来改变其一些细胞过程。拟议的研究的目的是确定有助于肿瘤细胞扩散的相关分子和细胞过程，以便临床干预可以针对那些关键的分子和过程。