Isolation and Characterization of Tumor Stem Cells from Melanoma Patients

黑色素瘤患者肿瘤干细胞的分离和表征

基本信息

批准号：
8616119
负责人：
ALEXANDER D BOIKO
金额：
$ 23.41万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-04-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8616119
关键词：
Affect Biological Assay Candidate Disease Gene Cell Count Cell Culture Techniques Cell Line Cell Separation Cell physiology Cell surface Cells Characteristics Data Development Dimensions Disease Distal Engraftment Environment Epidemic Epigenetic Process Frequencies Gene Expression Gene Expression Profile Gene Expression Profiling Genes Genetic Goals Human Immunophenotyping Knowledge Laboratories Malignant Neoplasms Medical Melanoma Cell Methods Molecular Molecular Profiling Mus Mutation Neoplasm Metastasis Neoplasm Transplantation Neoplasms Organ Pathogenesis Pathway interactions Patients Phenotype Plant Roots Population Property Radio Regimen Reporter Reporting Repression Resistance Role Sampling Site Skin Speed Staging Surface Antigens Technology Testing Therapeutic Tissues Tumor Stem Cells United States Visceral Xenograft Model base cancer cell cancer therapy design improved in vivo in vivo Model melanocyte melanoma molecular phenotype mouse model neoplastic cell protein expression reconstitution screening self-renewal small hairpin RNA stem stemness therapy design tumor tumor growth tumorigenic

项目摘要

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.

项目概要在缺乏有效治疗方案的情况下，美国黑色素瘤的迅速增加凸显了这一点深入了解这种疾病发病机制的紧迫性和重要性。在这个在该项目中，我们提出黑色素瘤是由 CD271 阳性肿瘤起始细胞亚群驱动的。瘤其他癌症中的干细胞（TSC）被证明对大多数传统化疗和放射具有抵抗力疗法，具有自我更新和有效补充消除的整个肿瘤细胞群的能力经过传统医疗方案后。因此，为了设计更成功的癌症疗法，必须进行分离和研究各个 TSC 的属性。该提案的主要目标是假定分离高度富集的来自广谱黑色素瘤的黑色素瘤 TSC 并表征其分子谱。我们将达到通过设计并完成以下实验任务来实现这一目标： 1. 通过 CD271 和其他细胞表面标志物表达识别并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测定。 2. 通过以下方式表征 CD271+ 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技术对 MTSC 进行全局基因表达分析；确认特定候选基因在 MTSC 表型中的关键作用 3. 分析MTSC在密切反映自然环境的先进致瘤特性通过表征 MTSC 的侵袭特征和功能作用来了解这种癌症在人类中的发生 CD271在人类皮肤重建小鼠模型中的应用。