Isolation and Characterization of Tumor Stem Cells from Melanoma Patients

描述（由申请人提供）：在缺乏有效治疗方案的情况下，黑色素瘤在美国迅速上升，这凸显了深入了解该疾病发病机制的紧迫性和重要性。在这个项目中，我们提出黑色素瘤是由CD271阳性肿瘤起始细胞亚群驱动的。其他癌症的肿瘤干细胞（TSCs）被证明对大多数常规化疗和放射治疗具有耐药性，具有自我更新的能力，并有效地补充传统医疗方案后消除的整个肿瘤细胞群。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的tsc的特性。本提案的主要目标是从广谱黑色素瘤中推定分离高度富集的黑色素瘤TSCs，并表征其分子谱。我们将通过设计和完成以下实验任务来实现这一目标：

DESCRIPTION (provided by applicant): 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.