Multiplexed Methods for the Study of Chromosomal Aberrations in Cancer

研究癌症染色体畸变的多重方法

• 批准号: 7193801
• 负责人: Aleksandar Milosavljevic
• 金额: $ 23.05万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7193801
• 关键词: Bacterial Artificial Chromosomes; Biological; Biological Assay; Biological Markers; Biological Process; Biological Sciences; Breast; Breast Cancer Cell; Cancer cell line; Cell Line; Chromosomal Rearrangement; Chromosome abnormality; DNA Sequence Rearrangement; Data; Databases; Detection; Development; Drug Delivery Systems; Epithelial Cells; Genes; Genome; Genomics; Human Genome; Informatics; Libraries; MCF7 cell; Malignant Neoplasms; Maps; Measures; Methods; Molecular; Online Systems; Phenotype; Preparation; Primary Neoplasm; Process; Recruitment Activity; Recurrence; Research Infrastructure; Research Personnel; Resolution; Robotics; Shotgun Sequencing; Surveys; Technology; Testing; Therapeutic Intervention; Tissues; Validation; base; design; expression cloning; fight against; improved; indexing; interest; malignant breast neoplasm; next generation; programs; research study; tumor; tumor progression

DESCRIPTION (provided by applicant): The general aim of the present application is to recruit newly available genomic technologies such as Bacterial Artificial Chromosome (BAC) library preparations, finished sequence of the human genome, and next- generation sequencing technologies developed by the 454 Life Sciences Corporation in the fight against cancer. This application aims to develop the Barcoded Tag Pooled Genomic Indexing (BT-PGI) method for highly multiplexed BAC-based mapping of chromosomal aberrations in cancer. The BT-PGI method achieves throughput and resolution by combining two types of multiplexing -- the pooling of BAC clones by the PGI method and the parallel sequencing of short mappable sequence tags by the newly developed 454 sequencing technology. A commercially available BAC library obtained from the well-established MCF-7 breast cancer cell line will be used for'testing and validation of these methods. Specific rearrangements detected in MCF-7 will then be measured in other breast cancer cell lines, and in breast cancer tissue. The biological function of these rearrangements will be tested by cloning and expression in both immortalized and transformed breast epithelial cells. Biological assays will depend upon the rearrangement or breakpoints of interest, but will focus on proliferation, survival, invasion and transformation. The improved understanding of chromosomal rearrangements will open new opportunities for charting the progression of cancer, understanding relevant molecular mechanisms, identifying biomarkers for informed therapy, and identifying targets for therapeutic intervention in breast cancer and other tumors.

描述（由申请人提供）：本申请的总体目的是招募新近可用的基因组技术，例如细菌人工染色体（BAC）文库制备、人类基因组的完成序列和由454生命科学公司开发的用于对抗癌症的下一代测序技术.本申请旨在开发条形码标签合并基因组索引（BT-PGI）方法，用于癌症中染色体畸变的高度多重BAC定位。BT-PGI方法通过结合两种类型的多路复用来实现通量和分辨率-通过PGI方法合并BAC克隆和通过新开发的454测序技术对短可映射序列标签进行平行测序。将使用从完善的MCF-7乳腺癌细胞系获得的市售BAC文库来测试和验证这些方法。在MCF-7中检测到的特异性重排将在其他乳腺癌细胞系和乳腺癌组织中进行测量。将通过在永生化和转化的乳腺上皮细胞中克隆和表达来测试这些重排的生物学功能。生物测定将取决于感兴趣的重排或断裂点，但将集中于增殖、存活、侵袭和转化。对染色体重排的更好理解将为绘制癌症进展图、理解相关分子机制、确定知情治疗的生物标志物以及确定乳腺癌和其他肿瘤治疗干预的靶点提供新的机会。