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Melanoma Metastasis Inhibition by Eristostatin

埃立他汀抑制黑色素瘤转移

基本信息

批准号：
6878142
负责人：
MARY ANN MCLANE
金额：
$ 21.52万
依托单位：
UNIVERSITY OF DELAWARE
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-04-01 至 2007-03-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Metastasis (i.e., tumor spread) is the major obstacle to cancer cure. The metastatic process consists of many steps. 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, has been found to inhibit melanoma metastasis in vivo using immunodeficient mice. To date, the basis for eristostatin's anti-metastatic effect remains unknown. The long term objective of this research is to identify molecular mechanisms by which melanoma metastasis may be inhibited. The specific goal proposed here is to investigate how eristostatin inhibits experimental metastasis by pursuing two specific aims: (i) Characterize the interactions of eristostatin and its mutants with four melanoma cell lines possessing distinct combinations of well-defined surface receptors. (ii) Identify the structural sequence within eristostatin which is most critical for its anti-metastatic ability. Recombinant eristostatin mutations will be created by alanine scanning mutagenesis, and made as bacterial fusion proteins with glutathione-S-transferase. These purified mutants will be used in a series of functional assays with four human metastatic melanoma cells lines. We will identify the specific molecule with which eristostatin interacts on each cell through crosslinking and immunological techniques. To determine which residue(s) of eristostatin are responsible for its cellular interactions, each mutant will be compared with wildtype eristostatin's activity in cellular assays. Those mutants which show the greatest difference in cellular interaction will be used in experimental metastasis assays. After i.v. injection of immunodeficient mice with melanoma cells and mutated eristostatin, mice will be observed for four weeks, and then necropsied. Metastatic potential will be assessed by counting lung metastases. Cryosections of lungs will be evaluated using immunohistochemical stains. Taken together, these studies will provide insights into how one naturally occurring protein possesses the "right fit" to bind melanoma cells and block their metastatic ability. This information will, in turn, lead to a rational design of therapeutic agents which would target these cells.

描述（由申请人提供）：转移（即，肿瘤扩散）是癌症治愈的主要障碍。转移过程包括许多步骤。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一员，已发现使用免疫缺陷小鼠在体内抑制黑色素瘤转移。迄今为止，蛇毒抑制素的抗转移作用的基础仍然未知。本研究的长期目标是确定抑制黑色素瘤转移的分子机制。这里提出的具体目标是研究如何通过追求两个具体目标来抑制实验转移：（i）表征蛇毒抑制素及其突变体与具有明确定义的表面受体的不同组合的四种黑素瘤细胞系的相互作用。(ii)确定对抗转移能力最关键的蛇毒蛋白的结构序列。通过丙氨酸扫描诱变将产生重组的蛇毒蛋白突变体，并将其制备为具有谷胱甘肽-S-转移酶的细菌融合蛋白。这些纯化的突变体将用于四种人转移性黑素瘤细胞系的一系列功能测定。我们将通过交联和免疫学技术鉴定与每个细胞相互作用的特定分子。为了确定蛇毒抑素的哪些残基负责其细胞相互作用，将在细胞测定中将每种突变体与野生型蛇毒抑素的活性进行比较。在细胞相互作用中显示最大差异的那些突变体将用于实验转移测定。在用黑色素瘤细胞和突变的蛇毒抑制素静脉注射免疫缺陷小鼠后，将观察小鼠四周，然后尸检。将通过计数肺转移瘤来评估转移潜力。将使用免疫组织化学染色评价肺冷冻切片。总的来说，这些研究将为了解一种天然存在的蛋白质如何具有“正确的适合”来结合黑色素瘤细胞并阻断其转移能力提供见解。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。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Scratching below the surface: wound healing and alanine mutagenesis provide unique insights into interactions between eristostatin, platelets and melanoma cells.

深入探究：伤口愈合和丙氨酸突变为了解毛立抑素、血小板和黑色素瘤细胞之间的相互作用提供了独特的见解。

DOI：
10.1159/000092417
发表时间：
2005
期刊：
Pathophysiology of haemostasis and thrombosis
影响因子：
0
作者：
McLane,MaryAnn;Zhang,Xiaoming;Tian,Jing;Zelinskas,Claire;Srivastava,Apoorva;Hensley,Brett;Paquette-Straub,Carrie
通讯作者：
Paquette-Straub,Carrie

A classification and nomenclature of disintegrins isolated from snake venoms.

从蛇毒中分离出的解整合素的分类和命名。