Gene expression profiling-based identification of cell-surface targets for developing multimeric ligands in pancreatic cancer.
Gene expression profiling-based identification of cell-surface targets for developing multimeric ligands in pancreatic cancer.
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DOI：
10.1158/1535-7163.mct-08-0402
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发表时间：
2008-09
影响因子：
5.7
通讯作者：
Han, Haiyong
中科院分区：
文献类型：
--
作者：
Balagurunathan, Yoganand;Morse, David L.;Hostetter, Galen;Shanmugam, Vijayalakshmi;Stafford, Phillip;Shack, Sonsoles;Pearson, John;Trissal, Maria;Demeure, Michael J.;Von Hoff, Daniel D.;Hruby, Victor J.;GillieS, Robert J.;Han, Haiyong
多聚体配体是含有同时靶向多个细胞表面蛋白的多个结合结构域的配体。由于协同结合，多聚体配体可以对表达所有靶蛋白的细胞（肿瘤）具有高亲合力，并且仅显示与不表达或仅表达一些靶的细胞（正常组织）的最小结合。鉴定在肿瘤细胞中同时表达而在正常细胞中不表达的靶点组合是一项具有挑战性的任务。在这里，我们描述了一种新的方法，用于确定这种组合使用全基因组基因表达谱，然后免疫组化。我们首先使用DNA微阵列生成了代表28种独特组织/细胞类型的28个胰腺癌标本和103个正常组织样本的mRNA基因表达谱数据库。然后从数据库中提取编码具有细胞表面表位的蛋白质的基因的表达数据，并使用新的基于多变量规则的计算方法进行分析，以鉴定在肿瘤中以有效结合水平表达的基因组合，但在正常组织中不表达。这些组合根据以有效水平表达集合的肿瘤样品的比例进一步排序。使用胰腺肿瘤组织和正常组织微阵列上的免疫组织化学证实了排名靠前的组合中所含基因的蛋白质表达。使用免疫细胞化学通过它们在胰腺癌细胞系中的组合表达进一步验证靶的共表达。因此，这些经验证的基因组合涵盖了一系列细胞表面靶点，可用于开发用于胰腺癌成像和治疗的多聚体配体。
Multimeric ligands are ligands that contain multiple binding domains that simultaneously target multiple cell surface proteins. Due to cooperative binding, multimeric ligands can have high avidity for cells (tumor) expressing all targeting proteins and only show minimal binding to cells (normal tissues) expressing none or only some of the targets. Identifying combinations of targets that concurrently express in tumor cells, but not in normal cells is a challenging task. Here, we describe a novel approach for identifying such combinations using genome-wide gene expression profiling followed by immunohistochemistry. We first generated a database of mRNA gene expression profiles for 28 pancreatic cancer specimens and 103 normal tissue samples representing 28 unique tissue/cell types using DNA microarrays. The expression data for genes that encode proteins with cell surface epitopes were then extracted from the database and analyzed using a novel multivariate rule-based computational approach to identify gene combinations that are expressed at an efficient binding level in tumors, but not in normal tissues. These combinations were further ranked according to the proportion of tumor samples that expressed the sets at efficient levels. Protein expression of the genes contained in the top ranked combinations was confirmed using immunohistochemistry on a pancreatic tumor tissue and normal tissue microarrays. Co-expression of targets was further validated by their combined expression in pancreatic cancer cell lines using immunocytochemistry. These validated gene combinations thus encompass a list of cell surface targets that can be used to develop multimeric ligands for the imaging and treatment of pancreatic cancer.