DIRECT INTERACTION OF ERISTOSTATIN WITH MELANOMA CELL SURFACE MOLECULES

埃立司他汀与黑色素瘤细胞表面分子的直接相互作用

• 批准号: 8359618
• 负责人: MARY ANN MCLANE
• 金额: $ 7.01万
• 依托单位: UNIVERSITY OF DELAWARE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8359618
• 关键词: 3-Dimensional; Adhesions; Affect; Atomic Force Microscopy; Binding; Cell Communication; Cell Line; Cell Survival; Cell physiology; Cell surface; Cells; Chemicals; Collagen; Confocal Microscopy; Delaware; Disintegrins; Environment; Event; Family; Fibronectins; Funding; Goals; Grant; Human; Lead; Lung; Malignant Neoplasms; Melanoma Cell; Molecular; Mus; National Center for Research Resources; Neoplasm Metastasis; Pathway interactions; Principal Investigator; Proteins; Research; Research Infrastructure; Resources; Signal Transduction; Source; Therapeutic Agents; United States National Institutes of Health; Venoms; base; cell growth; cell motility; cost; crosslink; design; insight; melanoma; member; metastatic process; response; tissue culture; tumor; two-dimensional

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Metastasis (i.e., tumor spread) is the major obstacle to cancer cure. The metastatic process consists of many steps, including those involving motility and adhesion. If the cascade of events is interrupted at any step, metastasis will not occur. Eristostatin, a member of the disintegrin family of viper venom proteins, can inhibit melanoma cell colonization in the lungs of mice injected with either murine or human melanoma cells. We have shown that the presence of eristostatin can cause changes in adhesion and motility in melanoma cells, but does not affect cell growth or survival. To date, the basis for eristostatin's anti-metastatic effect remains unknown, but these functional studies have opened the way to unlocking this natural protein's anti-cancer mechanism. The long term objective of this research is to identify mechanisms by which melanoma metastasis may be inhibited. The specific goals proposed here are to investigate the molecular mechanism used by eristostatin to affect melanoma cell function. We will do this by pursuing three specific aims: (i) Characterize the direct interaction of eristostatin with melanoma cell molecule(s). We will use atomic force microscopy, confocal microscopy and chemical crosslinking to identify the binding partner for eristostatin which is present on 6 different melanoma cell lines with varying degrees of metastatic potential. (ii) Compare the interaction differences between eristostatin and melanoma cells in a 2-dimensional versus 3-dimensional environment. Cellular responses can be very different in tissue culture (2-D) compared with what happens in the body (3-D). We will investigate eristostatin-melanoma cell interactions involving adhesion, proliferation and signaling in a 3-dimensional environment using matrices containing fibronectin and/or collagen. (iii) Identify intracellular signaling events associated with eristostatin's interaction with melanoma cells. The clearest example of melanoma cell activity inhibition by eristostatin is with motility when placed on fibronectin. We will perform immunochemical assessment of the intracellular signaling occurring when melanoma cells are exposed to eristostatin, focusing on known pathways associated with motility. Taken together, these studies will provide insights into how one naturally occurring protein, eristostatin, possesses the "right fit" to bind melanoma cells and block their metastatic ability. The information will, in turn, lead to a rational design of therapeutic agents which will target these cells.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 转移（即，肿瘤扩散）是癌症治愈的主要障碍。转移过程由许多步骤组成，包括涉及运动和粘附的步骤。如果事件级联在任何步骤中断，则不会发生转移。蛇毒蛋白去整合素家族的成员之一，能抑制黑色素瘤细胞在注射鼠或人黑色素瘤细胞的小鼠肺中的定植。我们已经证明，蛇毒蛋白的存在可以引起黑色素瘤细胞的粘附和运动的变化，但不影响细胞的生长或存活。到目前为止，eristostatin的抗转移作用的基础仍然未知，但这些功能性研究为解开这种天然蛋白质的抗癌机制开辟了道路。本研究的长期目标是确定抑制黑色素瘤转移的机制。本文提出的具体目标是研究蛇毒抑制素影响黑色素瘤细胞功能的分子机制。我们将通过追求三个具体目标来做到这一点：（i）表征蛇毒蛋白与黑素瘤细胞分子的直接相互作用。我们将使用原子力显微镜，共聚焦显微镜和化学交联，以确定结合伴侣的eristostatin这是目前在6个不同的黑色素瘤细胞系具有不同程度的转移潜力。 (ii)比较2维和3维环境中蛇毒蛋白和黑色素瘤细胞之间的相互作用差异。 组织培养（2-D）中的细胞反应与体内发生的反应（3-D）相比可能非常不同。我们将研究在三维环境中使用含有纤连蛋白和/或胶原蛋白的基质，涉及粘附，增殖和信号转导的蛇毒-黑色素瘤细胞相互作用。(iii)识别与蛇毒蛋白与黑色素瘤细胞相互作用相关的细胞内信号事件。 蛇毒抑制素抑制黑色素瘤细胞活性的最明显的例子是当置于纤连蛋白上时的运动性。我们将进行免疫化学评估的细胞内信号发生时，黑色素瘤细胞暴露于eristostatin，集中在已知的途径与运动。总之，这些研究将提供一种天然存在的蛋白质，蛇毒蛋白，如何拥有“正确的适合”结合黑色素瘤细胞和阻断其转移能力的见解。这些信息将反过来导致靶向这些细胞的治疗剂的合理设计。