Aptamer Based Technology for Molecular Analysis of Leukemia

描述（由申请人提供）：今天，数百万人患有癌症或患有癌症。为了对特定类型的癌症提供有效的治疗，癌症应该根据其分子特征进行分类。个体肿瘤的分子特征可用于定制治疗方案，监测对治疗的反应，并检测残留疾病。本研究的主要目标是开发一组适体作为白血病细胞分子谱分析的新型分子探针。基于适体的技术的最大优点是在所提出的研究中使用的独特的基于细胞的选择过程，Cell-SELEX（通过指数富集的配体系统进化）。一组细胞特异性适体可以使用扣除策略在相对短的时间内选择，甚至不知道哪些靶分子存在于细胞表面上。一旦选择了特定的适体，我们就可以开始对肿瘤细胞进行分析，而不需要识别每个单独的靶蛋白。这些类型的研究不能用其他类型的技术用于蛋白质组学研究。此外，所选择的适体不仅可以用作分子谱分析的分子探针，而且它们可以用作鉴定由肿瘤细胞或疾病状态中的其他细胞表达的新生物标志物的工具。核酸适体探针的稳定性和易合成性使其在制备蛋白质组学分子谱微阵列中具有比抗体更大的优势。因此，适体技术有望克服目前正常或肿瘤细胞分子谱分析中的挑战。我们开始我们的研究与良好的表征白血病细胞系开发基于细胞的SELEX方法。然后，我们将使用患者的白血病标本或肿瘤组织切片来选择能够识别患者白血病细胞或福尔马林固定的肿瘤细胞的适体。使用白血病细胞系或淋巴瘤肿瘤切片开发的策略应易于应用于其他癌细胞系或临床标本。在这个项目完成之前，我们将生产出用于不同类型白血病的适体探针组。通过流式细胞术或图像分析工具，细胞特异性适配子是用于肿瘤的分子诊断和分类以及用于评价药物对癌细胞的作用的有价值的探针，这反过来将改善癌症的诊断和治疗。本计画将利用细胞为基础的SELEX方法学，开发出可辨识白血病或淋巴瘤细胞的新型DNA适体探针。细胞特异性核酸适体是肿瘤分子诊断和分类的重要探针，也是评价药物对癌细胞作用的重要工具，有助于肿瘤的诊断和治疗。

DESCRIPTION (provided by applicant): Today, millions of people are living with cancer or have had cancer. To provide effective treatments for a particular type of cancer, cancers should be classified according to their molecular characteristics. The molecular characteristics of individual tumors can be used to tailor treatment regimens, monitor the response to treatment, and to detect residual diseases. The major goal of this proposal is to develop a panel of aptamers as novel molecular probes for molecular profiling of leukemia cells. The biggest advantage of the aptamer- based technology is the unique cell-based selection process, Cell- SELEX (Systematic Evolution of Ligands by Exponential enrichment), used in the proposed studies. A group of cell-specific aptamers can be selected using a subtraction strategy in a relatively short period without even knowing which target molecules are present on the cell surface. Once specific aptamers are selected, we can begin to profile tumor cells without identifying each individual target proteins. These types of studies cannot be done with other types of technologies used for proteonomic studies. Also, not only can the selected aptamers be used as molecular probes for molecular profiling, but they can be used as tools for identifying new biomarkers expressed by tumor cells or other cells in disease status. The stability and easy synthesis of DNA-aptamer probes will have many more advantages than antibodies in making microarrays for proteonomic molecular profiling. Thus, aptamer technology holds a promise to overcome the challenges in current molecular profiling of normal or tumor cells. We begin our studies with well-characterized leukemia cell lines to develop the cell-based SELEX methodologies. We are then going to use patients' leukemia specimens or tumor tissue sections to select aptamers that can recognize patients' leukemia cells or formalin-fixed tumor cells. The developed strategies using leukemia cell lines or lymphoma tumor sections should easily be applied to other cancer cell lines or clinical specimens. By the completion of this project, we will have produced panels of aptamer probes for different types of leukemias. With flow cytometry or tools of imaging analysis, the cell-specific aptamers are valuable probes for molecular diagnosis and classification of tumors, as well as for the evaluation of the effects of drugs on cancer cells, which in turn will improve the diagnosis and therapy of cancers. This project is to use cell-based SELEX methodologies for developing novel DNA aptamer probes that can recognize patients' leukemia or lymphoma cells. The cell-specific aptamers are valuable probes for molecular diagnosis and classification of tumors, as well as for the evaluation of the effects of drugs on cancer cells, which in turn will improve the diagnosis and therapy of cancers.