Breast Cancer Proteomic via Laser-Free Microdissection and Gemini Technologies

通过无激光显微切割和 Gemini 技术进行乳腺癌蛋白质组学

基本信息

批准号：
7290319
负责人：
Brian M Balgley
金额：
$ 15.26万
依托单位：
CALIBRANT BIOSYSTEMS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-09-22 至 2009-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7290319
关键词：
Abnormal Cell Adopted Adoption American Joint Committee on Cancer Area Automobile Driving Axillary lymph node group Back Behavior Biochemical Bioinformatics Biological Biological Assay Biological Markers Biology Biopsy Breast Breast Carcinoma Cancerous Carcinoma Cell Extracts Cells Chemicals Classification Clinical Clinical Course of Disease Collaborations Collection Complex Consumption Coupled DNA copy number Data Detection Development Diagnosis Diagnostic Diagnostic Neoplasm Staging Disease Distant Metastasis Ductal Epithelium Early Diagnosis Electrospray Ionization Environment Epidermal Growth Factor Receptor Estrogen Receptors Evaluation Event Fingerprint Freezing Functional disorder Gel Generations Genes Genetic Transcription Goals Heating Heterogeneity Human Human Genome Immunoassay Individual Information Management Institutes Ions Isotopes Laboratories Lasers Lesion Liquid Chromatography Loss of Heterozygosity Malignant Neoplasms Mammary Gland Parenchyma Manuals Mass Spectrum Analysis Measurement Methods Microdissection Molecular Molecular Abnormality Molecular Profiling Monitor Monoclonal Antibodies Morphologic artifacts Mucinous Nature Nodal Normal Cell Numbers Pathology Patients Pattern Peptides Performance Pharmaceutical Preparations Polymorphism Analysis Population Post-Translational Protein Processing Preclinical Drug Evaluation Predictive Factor Preparation Process Progesterone Receptors Protein Analysis Proteins Proteome Proteomics RNA Range Reagent Relative (related person)Reporting Reproducibility Research Role playing therapy Sampling Screening procedure Severities Shotguns Signal Transduction Single Nucleotide Polymorphism Specimen Staging Staining method Stains Standards of Weights and Measures Stroke Structure System Techniques Technology Testing Time Tissue Fixation Tissue Sample Tissue Stains Tissues Tumor Markers Tumor Pathology Two-Dimensional Polyacrylamide Gel Electrophoresis Universities Validation Variant Visual Work anticancer research base breast lesion cDNA Arrays cancer cell cancer proteomics cancer type cell type clinical phenotype comparative comparative genomic hybridization ductal breast carcinoma experience instrumentation laser capture microdissection malignant breast neoplasm medical schools neoplastic cell nervous system disorder novel professor prognostic protein expression receptor response size surface enhanced laser desorption ionization tool tumor two-dimensional

项目摘要

DESCRIPTION (provided by applicant): The generation of biologically relevant proteomics data requires samples consisting of homogenous cell populations, in which no unwanted cells of different types and/or development stages obscure the results. The problem is compounded for the analysis of tissue biopsies, since many different cell types are typically present, and small numbers of abnormal cells may lie within or adjacent to unaffected areas. While methods such as laser capture microdissection (LCM) enable the isolation of homogeneous subpopulations of cells, proteomic analysis of LCM-procured specimens is severely constrained by the very low amounts of sample generated. To avoid the limitations of established proteome techniques for analyzing protein extracts obtained from microdissection-procured tissue specimens, an effective discovery-based proteome platform has recently been developed at Calibrant. This proteome platform, called Gemini, combines a unique multidimensional separation system with customized back-end bioinformatics tools, and allows ultrasensitive analysis of minute protein amounts extracted from cells captured by tissue microdissection. This project further aims to employ a novel, laser-free microdissection technique pioneered by our collaborator, Dr. Zhengping Zhuang at the National Institute of Neurological Disorders and Stroke (NINDS), capable of providing enriched, high quality, and reproducible tissue samples. By combining Calibrant's ability to perform proteomic profiling from minute samples with the technology and expertise offered by Dr. Zhuang (NINDS) in tissue microdissection and tumor pathology, the proposed research represents a synergistic effort toward the evaluation and validation of a novel biomarker discovery paradigm for enabling the proteome analysis of cancer cells and their micro-environment in support of cancer research, diagnosis, and treatment. Application of the resulting biomarker discovery platform for studying the molecular mechanisms associated with breast carcinoma at the global level will be realized through a collaboration with Professor Fattaneh A. Tavassoli (Yale University School of Medicine), who will apply more than 30 years of research experience in breast cancer pathology and biology and provide access to a collection of fresh frozen human breast cancer biopsies for biomarker discovery.

描述（由申请人提供）：生物相关蛋白质组学数据的生成需要由同质细胞群组成的样品，其中没有不同类型和/或发育阶段的不需要的细胞掩盖结果。对于组织活检的分析来说，问题变得更加复杂，因为通常存在许多不同的细胞类型，并且少量的异常细胞可能位于未受影响的区域内或邻近未受影响的区域。虽然激光捕获显微切割 (LCM) 等方法能够分离同质的细胞亚群，但 LCM 获得的样本的蛋白质组分析受到生成样品量极少的严重限制。为了避免现有蛋白质组技术在分析从显微切割获得的组织样本中获得的蛋白质提取物时的局限性，Calibrant 最近开发了一个有效的基于发现的蛋白质组平台。这个蛋白质组平台称为 Gemini，将独特的多维分离系统与定制的后端生物信息学工具相结合，并允许对从组织显微切割捕获的细胞中提取的微小蛋白质量进行超灵敏分析。该项目的进一步目标是采用由我们的合作者、国家神经疾病和中风研究所 (NINDS) 的庄正平博士首创的新型无激光显微切割技术，该技术能够提供丰富、高质量和可重复的组织样本。通过将 Calibrant 从微小样本中进行蛋白质组分析的能力与 Zhuang 博士 (NINDS) 在组织显微切割和肿瘤病理学方面提供的技术和专业知识相结合，拟议的研究代表了对新型生物标志物发现范式的评估和验证的协同努力。能够对癌细胞及其微环境进行蛋白质组分析，以支持癌症研究、诊断和治疗。通过与 Fattaneh A. Tavassoli 教授（耶鲁大学医学院）合作，将应用由此产生的生物标志物发现平台在全球范围内研究与乳腺癌相关的分子机制，该教授将运用 30 多年的研究经验乳腺癌病理学和生物学领域的研究人员，并提供一系列新鲜冷冻人类乳腺癌活检样本以发现生物标志物。