A chemical proteomics approach for the identification of accessible antigens expressed in human kidney cancer
A chemical proteomics approach for the identification of accessible antigens expressed in human kidney cancer
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DOI：
10.1074/mcp.m600164-mcp200
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发表时间：
2006-11-01
影响因子：
7
通讯作者：
Neri, Dario
中科院分区：
文献类型：
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作者：
Castronovo, Vincent;Waltregny, David;Neri, Dario
开发更具选择性的抗癌药物的一个有希望的途径是，通过结合肿瘤相关标记物的特异性分子，将生物活性分子定向输送到肿瘤环境中。我们使用了一种化学蛋白质组学方法，该方法基于手术切除的患有肿瘤的人肾脏内可访问结构的体外灌流和生物素化，以获得关于可访问和丰富的抗原的信息，这些抗原在人类癌症中过度表达。这一过程导致了血管结构的生物素选择性标记。生物素化蛋白质经链霉亲和素树脂纯化后，用质谱学方法鉴定，共发现637个蛋白质，其中184个仅在肿瘤标本中发现，223个仅在正常肾脏的部分组织中发现。免疫组织化学和聚合酶链式反应分析证实，这项研究中确定的几种可能的癌症抗原确实在肿瘤中优先表达。总之，我们开发了一种方法，可以在人体组织中识别可访问的生物标记物。这项研究中确定的肿瘤相关抗原可能是基于抗体的抗癌治疗的合适靶点。这里描述的实验方法应该适用于其他外科标本和其他病理学，以及基本的生理和免疫过程的研究。
A promising avenue toward the development of more selective anticancer drugs consists in the targeted delivery of bioactive molecules to the tumor environment by means of binding molecules specific to tumor-associated markers. We have used a chemical proteomics approach based on the ex vivo perfusion and biotinylation of accessible structures within surgically resected human kidneys with tumor to gain information about accessible and abundant antigens that are overexpressed in human cancer. This procedure led to the selective labeling with biotin of vascular structures. Biotinylated proteins were purified on streptavidin resin and identified using mass spectrometric methodologies, revealing 637 proteins, 184 of which were only found in tumor specimens and 223 of which were only found in portions of normal kidneys. Immunohistochemical and PCR analysis confirmed that several of the putative cancer antigens identified in this study are indeed preferentially expressed in tumors. In conclusion, we have developed a methodology that allows the identification of accessible biomarkers in human tissues. The tumor-associated antigens identified in this study may be suitable targets for antibody-based anticancer therapies. The experimental approach described here should be applicable to other surgical specimens and to other pathologies as well as to the study of basic physiological and immunological processes.