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: 7668066
• 负责人: JAMES P QUIGLEY
• 金额: $ 39.32万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-05 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7668066
• 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）：肿瘤传播的许多具体步骤已被广泛建模和分子解剖。然而，转移级联中的一个早期步骤，即内渗，即逃逸的肿瘤细胞进入脉管系统，目前的研究相对较少。原因之一是迄今为止，尚未可用于比较分析在其内渗能力方面表现出显着差异的人类肿瘤细胞表型变异。我们的实验室报告了人类纤维肉瘤细胞系 (HT-1080) 的两种变体的选择和分离，它们的内渗率显着不同（50-100 倍），从而在转移能力方面产生了类似的 > 50 倍差异。这两种肿瘤传播变体HT-hi/diss和HT-lo/diss是在鸡胚胎模型中选择和监测的，其中在胚胎绒毛尿囊膜（CAM）上发育的原发性人类肿瘤再现了多步骤的肿瘤传播过程并在许多次要器官中形成微转移灶。我们还在不同的小鼠转移试验中测试了两种纤维肉瘤变体，并证实了它们的肿瘤传播能力的显着差异。因此，这两种同源的内渗变异适用于早期转移事件的比较分析。因此，在具体目标 1 中，我们建议从分子角度剖析决定肿瘤细胞内渗事件结果的三个生理/病理过程，即诱导血管生成、基质侵袭和向血管性。我们将分析选定分子对肿瘤细胞内渗相关事件的功能作用，包括：炎症细胞MMP-9； FGF-2；尿素PA；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识别参与促血管事件的贡献分子。肿瘤细胞传播和血管生成的小鼠模型将用于补充鸡胚模型，以检查两种纤维肉瘤变体中已识别的分子。然而，明显缺乏其他肿瘤细胞内渗变异可供比较和对比，这与癌症（人类的主要癌症）尤其相关。因此，对于具体目标 2，我们建议通过采用体内选择的不同内渗率的癌变体来鉴定在机制上有助于癌症传播内渗步骤的特定蛋白质。我们将从人类前列腺癌细胞系、结肠癌细胞系和胰腺癌细胞系中产生成对的高和低血管内变体。功能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变体中鉴定的细胞机制和贡献分子。间充质成纤维细胞及其产物对癌变体传播特性的影响也将被量化。有关癌细胞内渗的新机制信息将为肿瘤传播的特定步骤（即内渗）提供分子联系。公共卫生相关性：癌症导致的死亡主要是因为肿瘤细胞从原发肿瘤部位扩散到其他重要器官和组织。肿瘤通常通过脉管系统扩散。为了使肿瘤细胞逃离原发肿瘤并进入脉管系统，它们通过产生不同水平的功能分子来改变一些细胞过程。拟议研究的目标是确定有助于肿瘤细胞扩散的相关分子和细胞过程，以便临床干预能够针对这些关键分子和过程。