BAC-FISH Assays for Sensitive Karyotyping of Cancer Cells

用于癌细胞敏感核型分析的 BAC-FISH 检测

基本信息

批准号：
7489852
负责人：
Heinz-Ulrich Guenter Weier
金额：
$ 20.62万
依托单位：
UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-01 至 2010-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7489852
关键词：
Address Aging Bacterial Artificial Chromosomes Behavior Benign Binding Biological Assay Cancer Patient Cancer cell line Case Study Cations Cells Characteristics Chromosomal Rearrangement Chromosomal translocation Chromosome Band Chromosome Banding Chromosome Painting Chromosome abnormality Chromosomes Chromosomes, Human, Pair 13 Chromosomes, Human, Pair 7 Clinical Code Collection Color Complex Condition Coupled Couples Cytogenetic Analysis Cytogenetics DNA DNA Probes DNA Sequence Rearrangement Detection Development Diagnosis Diagnostic Neoplasm Staging Disease Disease Progression Familial disease Fertility Fluorescence Microscopy Fluorescent in Situ Hybridization Frequencies G-Banding Gene Amplification Genes Genome Genomics Homologous Gene Human Human Genome Image In Situ Hybridization Individual Infertility Karyotype Karyotype determination procedure Knowledge Label Laboratories Lead Lesion Libraries Malignant Neoplasms Maps Measures Meiosis Mental Retardation Metaphase Methods Microscope Molecular Cytogenetics Mutation Neoplasm Metastasis Neoplasms Occupational Outcome Patients Personal Satisfaction Phenotype Population Positioning Attribute Predisposition Premalignant Procedures Proteins Publishing Reagent Reproducibility Research Research Personnel Research Project Grants Resolution Scheme Screening procedure Services Slide Specificity Specimen Spectral Karyotyping Support of Research System Techniques Technology Testing Tissues Trisomy Tumor Cell Line Tumor stage Work anticancer research base cancer cell cost genotoxicity improved innovation neoplastic neoplastic cell novel response size success technology development tumor tumor progression

项目摘要

DESCRIPTION (provided by applicant): The detailed cytogenetic analyses of human tumors and cancer tissues, in particular, have revealed the presence of structural chromosome abnormalities in most of cases. The underlying changes are often specific for the type of tumor or cancer and the cells' altered phenotype. Furthermore, investigators could demonstrate a correlation between the type or extent of chromosome changes, disease progression, and outcome. However, present laboratory techniques for the screening for structural abnormalities are very limited with respect to the detection of small, 'cryptic' translocations. For example, Giemsa (G)-banding of metaphase chromosomes or the fluorescence in situ hybridization (FISH)-based techniques of whole chromosome painting (WCP) and Spectral Karyotyping (SKY) analysis typically miss translocations that involve segments of less than 10 megabasepairs (Mbp), i.e., about the size of a chromosome band. We postulate that small cryptic translocations exist undetected in the genomes of individuals with a normal phenotype or diseases such as mental retardation, impaired fertility, precancerous lesions, or early stage tumors. Knowledge about structural alterations and chromosomal imbalances might help clinicians make more accurate predictions regarding the onset and course of a disease. This R21 project will investigate the feasibility to rapidly and inexpensively screen the human genome for the presence of occult cryptic translocations (OCTs). Specifically, we will develop and test FISH assays using collections of validated chromosome-specific bacterial artificial chromosomes (BACs) for the detection of OCTs in human cancer cells. With BAC probes spaced on average 0.9 Mbp apart and covering the entire euchromatic part of the human genome, we expect our 'BAC-FISH' assay to lead to greatly increased sensitivity compared to WCP or banding tests. Our innovative assay for sensitive genome-wide screening for translocations will be developed with cancer cell lines for which limited information about structural abnormalities is available. At the end of this project, we will be well positioned to conduct a larger study of the frequency of OCTs in the normal population as well as tumor cells and to offer BAC-FISH screening service and reagents to research and clinical laboratories for collaborative studies.

描述（由申请人提供）：尤其是人类肿瘤和癌症组织的详细细胞遗传学分析显示，在大多数情况下，存在结构性染色体异常。潜在的变化通常是针对肿瘤或癌症的类型和细胞改变的表型的特异性变化。此外，研究人员可以证明染色体变化，疾病进展和结果的类型或程度之间存在相关性。但是，对于检测小的“隐秘”易位，目前用于筛查结构异常的实验室技术非常有限。例如，GIEMSA（G） - 中期染色体或原位杂交（FISH）基于整个染色体绘画（WCP）的基于原位杂交（FISH）的技术（SKY）分析（SKY）分析通常会错过易位（SKY）分析（SKY）分析（通常会涉及少于10兆巴epairairairairs的易变） MBP），即大约染色体带的大小。我们假设在具有正常表型或疾病的个体的基因组中未发现小的隐性易位，例如智力低下，生育能力受损，癌前病变或早期肿瘤。关于结构改变和染色体失衡的知识可能有助于临床医生对疾病的发作和过程做出更准确的预测。这个R21项目将调查快速和廉价地筛选人类基因组的可行性，以实现神秘的隐性易位（OCT）。具体而言，我们将使用经过验证的染色体特异性细菌人工染色体（BAC）的收集来开发和测试鱼类测定法，以检测人类癌细胞中的OCT。由于BAC探针平均分开0.9 MBP并覆盖了人类基因组的整个正念部分，因此与WCP或频段测试相比，我们的“ BAC-FISH”测定法会大大提高灵敏度。我们对易感基因组筛查的创新分析将使用癌细胞系进行开发，以提供有关结构异常信息的有限信息。在该项目结束时，我们将有能力对正常人群以及肿瘤细胞中OCT的频率进行更大的研究，并为研究和临床实验室提供协作研究的BAC-FISH筛查服务和试剂。