Isolation and Characterization of Tumor Stem Cells from Melanoma Patients

项目摘要 黑色素瘤在美国的迅速上升，在缺乏有效的治疗方案强调， 进一步认识本病发病机制的紧迫性和重要性。在这 我们提出黑色素瘤是由CD 271阳性肿瘤起始细胞亚群驱动的。肿瘤 其他癌症中的干细胞（TSCs）对大多数常规化疗和放疗都有抵抗力， 治疗，有能力自我更新和有效地补充整个肿瘤细胞群消除 在传统的医疗方案之后。因此，为了设计更成功的癌症治疗方法，必须分离和研究 各自TSC的特性。该提案的主要目标是将高度浓缩的 从广谱黑色素瘤中分离黑色素瘤TSC并表征其分子谱。我们将达到 设计并完成了以下实验任务： 1.通过CD 271和其他细胞表面标志物表达识别和前瞻性分离黑色素瘤TSC 使用先进的荧光激活细胞分选和体内肿瘤移植试验。 2.通过以下方式表征位于CD 271 + MTSC功能根源的遗传和/或表观遗传改变 使用微阵列和慢病毒报告基因技术进行MTSC的整体基因表达分析; 证实特定候选基因在MTSC表型中的关键作用 3.分析MTSC在密切反映自然环境的环境中的高级致瘤性 通过描述MTSC的侵袭性特征和功能作用， CD 271在人皮肤中的表达重建小鼠模型。

Project Summary The rapid rise of melanoma, in the United States, in the absence of effective therapeutic regimens underscores the urgency and importance of obtaining a deeper knowledge about pathogenesis of this disease. In this project we propose that melanoma is driven by subpopulation of CD271 positive tumor initiating cells. Tumor stem cells (TSCs) in other cancers were shown to be resistant to most conventional chemo and radio therapies, to have the ability to self-renew and efficiently replenish the entire tumor cell population eliminated after traditional medical regimens. Thus to design more successful cancer therapy one must isolate and study the properties of respective TSCs. The primary goal of this proposal is to putatively isolate highly enriched melanoma TSCs from broad spectrum of melanomas and to characterize their molecular profile. We will reach this goal by designing and accomplishing following experimental tasks: 1. Identify and prospectively isolate melanoma TSC by CD271 and additional cell surface markers expression using advanced Fluorescent Activated Cell Sorting and in-vivo tumor transplantation assays. 2. Characterize genetic and/or epigenetic alterations that lie at the root of CD271+ MTSC function by performing global gene expression analysis of MTSCs using microarray and lentiviral reporter technologies; confirm critical role of specific gene candidates in MTSC phenotype 3. Analyze MTSC's advanced tumorigenic properties in the environment that closely reflects natural occurrence of this cancer in humans by characterizing invasive characteristics of MTSCs and a functional role of CD271 in human skin reconstruct mouse model.