Single Cell Technologies for Rapid Detection of Tumor Heterogeneity

用于快速检测肿瘤异质性的单细胞技术

基本信息

批准号：
9133323
负责人：
Karen Sue Anderson
金额：
$ 23.81万
依托单位：
ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-01 至 2018-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9133323
关键词：
Aneuploidy Binding Bioinformatics Biological Assay Biological Markers Breast Cancer Cell Breast Cancer cell line Cell Separation Cells Combined Modality Therapy Complementary DNA Copy Number Polymorphism DNA Data Development Diploidy Dissection Evolution Excision Flow Cytometry Frequencies Genes Genomics Geometry Health Heterogeneity Human Individual Link Malignant - descriptor Malignant Neoplasms Mammary Neoplasms Measures Messenger RNA Methods Modification Molecular Molecular Analysis Monitor Mutate Mutation Mutation Analysis Mutation Detection Nanostructures Natural regeneration PIK3CA gene PTEN gene Ploidies Population Population Heterogeneity Primary Neoplasm Relapse Reproducibility Resistance Risk Sensitivity and Specificity Site Solid Neoplasm Somatic Mutation Sorting - Cell Movement Specificity TP53 gene Targeted Resequencing Technology Testing Therapeutic Time Tissues Transfection Translations Tumor Cell Line Tumor Suppressor Genes Tumor Suppressor Proteins anticancer research base cancer cell clinically relevant cost effective deep sequencing design established cell line flexibility improved innovation malignant breast neoplasm neoplastic cell next generation novel prevent rapid detection response scaffold single cell technology targeted treatment technology development therapy resistant tool tumor tumor heterogeneity tumor progression

项目摘要

DESCRIPTION (provided by applicant): Currently, there are few methods available to analyze the evolution of tumor heterogeneity; micro-dissection of tumors only provides information on major species of malignant cells but is unable to detect rare therapy-resistant subclones that have the potential to regenerate tumors. Identification of these rare cells by single-cell isolatio and sequencing is both time-consuming and prohibitively expensive. Recent next-generation deep sequencing studies have demonstrated the clinical relevance of clonal heterogeneity within individual cancers, but currently rapid and cost-effective methods to measure and track the rates of co-occurrences of mutations in cell populations do not exist. Therefore, the development of rapid, flexible, single- cell technologies with the capacity to identify heterogeneous mutations of multiple genes in individual cells within bulk populations is critical for the development of effective targeted therapies that prevent tumor relapse. To overcome this challenge, we propose to adapt novel nanomolecular scaffolds (termed "DNA origami") to transfect tumor cells and capture mRNA encoding known-tumor suppressor genes. We propose to test the specificity of these scaffolds in breast tumor cell lines and primary breast tumor with known mutations in p53, PTEN, and PIK3CA genes. This approach will allow the rapid quantitation of genomic diversity and evolutionary order in cells from solid tumors for improved targeting of rare malignant subclones.

描述（由适用提供）：目前，几乎没有可用的方法来分析肿瘤异质性的演变；肿瘤的微截止性仅提供有关主要种类恶性细胞的信息，但无法检测到具有再生肿瘤潜力的罕见耐药亚克隆。通过单细胞隔离和测序鉴定这些稀有细胞既耗时又是昂贵的。最近的下一代深度测序研究表明，单个癌症内克隆异质性的临床相关性，但是目前尚不存在测量和跟踪细胞种群突变共发生率的快速且具有成本效益的方法。因此，开发快速，灵活的单细胞技术具有鉴定大量种群中单个细胞中多个基因的异质突变的能力，对于开发有效的靶向疗法以防止肿瘤缓解。为了克服这一挑战，我们建议适应新型的纳米分子支架（称为“ DNA折纸”），以翻译肿瘤细胞并捕获编码已知肿瘤抑制基因的mRNA。我们建议在乳腺肿瘤细胞系和原发性乳腺肿瘤中测试这些支架的特异性 p53，PTEN和PIK3CA基因中的已知突变。这种方法将允许从实体瘤细胞中快速定量基因组多样性和进化顺序，以改善靶向罕见的恶性亚克隆的靶向。