Phage Display to Identify Epithelial to Mesenchymal Transitioned (EMT) Breast Cancer Cells

 描述(申请人提供)：这项研究的主要目标是确定上皮间充质转化(EMT)乳腺癌细胞的探针/标记物。癌细胞上皮间充质转化是上皮型癌细胞侵袭、转移到远处器官所表现出的重要现象。这是一个过程 上皮特征的丧失和癌细胞间充质样表型的获得。它被认为是肿瘤细胞扩散到周围组织和循环的先行者和使者。然而，由于用于定义这些转化细胞的间质标记物的非特异性，对这些转化细胞的表征一直是具有挑战性的。因此，EMT和随后的侵袭的图景相对不清楚，需要一组更雄心勃勃的特定标记来识别这些转化的细胞群体，并将它们与其他间充质细胞如成纤维细胞区分开来。在这个概念框架内，一个新的挑战是找到一组与这一过程相关的特定生物标志物。我们提出了一项新颖和创新的研究，利用噬菌体展示文库来鉴定能够在体外与人乳腺癌组织中EMT转移的乳腺癌细胞特异性和选择性结合的噬菌体，并帮助将它们与周围的成纤维细胞区分开来。整个研究旨在获得与EMT特异性乳腺癌细胞结合的噬菌体配体。该项目将包括：1)筛选与乳腺癌相关成纤维细胞(CAF‘s)消减筛选后与人转移性乳腺癌细胞组具有高亲和力、选择性和特异性的噬菌体。2)测试所选择的噬菌体与EMT诱导的乳腺癌细胞模型的结合；以及3)测试所选择的噬菌体与人乳腺癌组织(外膜)的结合。该项目的成功完成将导致新一代噬菌体衍生探针的开发，这种探针可进一步用于诊断、治疗、分析和成像。

 DESCRIPTION (provided by applicant): The main goal of this study is to identify probes/markers for epithelial to mesenchymal transformed (EMT) breast cancer cells. Epithelial to Mesenchymal transition of cancer cells is a crucial phenomenon demonstrated by the epithelial type cancer cells in order to invade, and metastasize to distant organs. It is a process of loss of epithelial characteristics and the acquisition of a mesenchymal-like phenotype of cancer cells. It is thought to precede and enable the dissemination of tumor cells into the surrounding tissue and circulation. However characterization of these transformed cells has been challenging because of the non-specificity of the mesenchymal markers used to define them. So the picture of EMT and subsequent invasion is relatively unclear and warrants a more ambitious set of specific markers that can identify these transformed cell population and distinguish them from other mesenchymal cells such as fibroblasts. A new challenge, within this frame of concept, is to find specific set of biomarkers relevant for this process. We propose a novel and innovative study to use phage display libraries for identification of phages that can specifically and selectively bind to the EMT transitioned breast cancer cells invitro, in human breasts cancer tissues and help to distinguish them from the surrounding fibroblasts. This whole study is directed to obtain phage ligands binding to EMT-specific breast cancer cells. The project will include: 1) Selection and characterization phages that bind with high affinity, selectivity and specificity to the panel of human metastatic breast cancer cells after subtractive screening against mammary carcinoma- associated fibroblasts (CAF's). 2) Testing the binding of the selected phages to EMT-induced models of breast cancer cells and 3) Testing the binding of the selected phages to human breast cancer tissue (exvivio). Successful accomplishment of this project would lead to the development of the novel generation of phage-derived probes that can be further used for diagnostics, therapy, profiling and imaging.