Aptamer Based Technology for Molecular Analysis of Leukemia

描述（由申请人提供）：今天，数以百万计的人患有癌症或曾经患过癌症。为了对特定类型的癌症提供有效的治疗，癌症应该根据它们的分子特征进行分类。单个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。本提案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势是独特的基于细胞的选择过程，Cell- SELEX（配体的系统进化指数富集），在拟议的研究中使用。一组细胞特异性适配体可以使用减法策略在相对较短的时间内选择，甚至不知道哪些靶分子存在于细胞表面。一旦选定了特定的适体，我们就可以开始分析肿瘤细胞，而无需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作为分子探针用于分子谱分析，而且可以用作鉴定肿瘤细胞或其他疾病状态细胞表达的新生物标志物的工具。与抗体相比，dna适体探针的稳定性和易于合成在制备用于蛋白质组学分子分析的微阵列方面具有更多的优势。因此，适体技术有望克服当前正常细胞或肿瘤细胞分子谱分析的挑战。我们开始研究具有良好特征的白血病细胞系，以开发基于细胞的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肿瘤细胞的适配体。使用白血病细胞系或淋巴瘤肿瘤切片的发展策略应该很容易应用于其他癌细胞系或临床标本。该项目完成后，我们将为不同类型的白血病生产适配体探针。利用流式细胞术或成像分析工具，细胞特异性适配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探针，从而提高癌症的诊断和治疗水平。这个项目是使用基于细胞的SELEX方法开发新的DNA适体探针，可以识别患者的白血病或淋巴瘤细胞。细胞特异性适配体是肿瘤分子诊断和分类的重要探针，也是评价药物对肿瘤细胞作用的重要探针，从而提高肿瘤的诊断和治疗水平。

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.