Isolation and Characterization of Tumor Stem Cells from Melanoma Patients

项目摘要 在缺乏有效治疗方案的情况下，黑色素瘤在美国的迅速上升突显了这一点 了解该病发病机制的紧迫性和重要性。在这 我们认为黑色素瘤是由CD271阳性的肿瘤启动细胞亚群驱动的。肿瘤 其他癌症中的干细胞(TSCs)被证明对大多数常规化疗和放射治疗具有抵抗力。 治疗，有能力自我更新和有效地补充整个消除的肿瘤细胞群 在传统的医疗方法之后。因此，要设计出更成功的癌症治疗方法，就必须分离和研究 各个TSC的属性。这项提议的主要目标是假定分离高度浓缩的 从广泛的黑色素瘤中分离黑色素瘤TSCs，并鉴定其分子特征。我们会到达 通过设计和完成以下实验任务来实现这一目标： 1.通过CD271和附加细胞表面标志物的表达鉴定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.通过以下方式描述CD271+MTSC功能根源的遗传和/或表观遗传学变化 使用微阵列和慢病毒报告技术进行MTSCs的全球基因表达分析； 确认特定基因候选在MTSC表型中的关键作用 3.在与自然环境密切相关的环境中分析MTSC的高级致癌特性 通过表征MTSCs的侵袭特性和功能作用研究这种癌症在人类中的发生 CD271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.