Ultrasensitive Approaches for Complex Animal Proteomics

复杂动物蛋白质组学的超灵敏方法

基本信息

批准号：
8056788
负责人：
Xueping Fang
金额：
$ 42.29万
依托单位：
CALIBRANT BIOSYSTEMS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-02-01 至 2013-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8056788
关键词：
Abnormal Cell Address Anaplastic astrocytoma Animals Archives Area Astrocytoma Automobile Driving Back Behavior Biochemical Bioinformatics Biological Biological Markers Biological Markers Biological Products Biopsy Biotechnology Blood capillaries Brain Businesses Cells Chromatography Clinical Clinical Trials Collaborations Complex Coupling Critical Pathways DNA DNA copy number Data Deposition Detection Detergents Development Diagnosis Diagnostic Disease Disease Marker Disease Progression Drug Approval Processes Electrophoresis Environment Epithelial Evaluation Event Foundations Freezing Functional disorder Gel Genetic Transcription Glioblastoma Glioma Goals Heterogeneity Human Human Genome Hydrophobicity Immunohistochemistry Individual Industry Investigation Ions Isoelectric Focusing Label Lesion Licensing Liquid Chromatography Loss of Heterozygosity Malignant Neoplasms Mass Spectrum Analysis Measurement Medical Membrane Membrane Proteins Methods Microdissection Mining Molecular Molecular Abnormality Molecular Profiling Monitor National Institute of Neurological Disorders and Stroke Nature Normal Cell Organ Ovarian Carcinoma Pathogenesis Peptide Hydrolases Peptides Performance Pharmaceutical Preparations Pharmacologic Substance Phase Phenotype Plasma Polymorphism Analysis Population Positioning Attribute Post-Transcriptional Regulation Post-Translational Protein Processing Preclinical Testing Preparation Process Progress Reports Proteins Proteome Proteomics Provider RNA Relative (related person)Reporting Reproducibility Research Sampling Series Serum Albumin Single Nucleotide Polymorphism Specimen Spectrometry, Mass, Electrospray Ionization Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Staging Structure Surveys System Techniques Technology Tissue Banks Tissue Sample Tissues Two-Dimensional Polyacrylamide Gel Electrophoresis United States Food and Drug Administration Validation Variant Western Blotting Yeasts base biological systems cDNA Arrays cancer cell capillary cell type clinically relevant commercialization comparative comparative genomic hybridization drug development drug discovery expectation gel electrophoresis improved instrumentation knowledge base laser capture microdissection nano neoplastic cell neuropathology novel phase 1 study product development prognostic protein aminoacid sequence protein complex protein expression public health relevance response success symposium technology development technology validation tool tumor

项目摘要

DESCRIPTION (provided by applicant): Biologically and clinically relevant proteomic data can only be generated if organ or tissue samples investigated consist of homogeneous cell populations, in which no unwanted cells of different types and/or development stages obscure the results. One of the main problems with the analysis of tissue samples is the heterogeneous nature of the sample. Many different cell types are typically present in tissue biopsies, and in the case of diseased tissue small numbers of abnormal cells may lie within or adjacent to unaffected areas. Thus, the laser capture/microdissection process has been developed to provide a rapid and straightforward method for procuring homogeneous subpopulations of cells or structures for biochemical and molecular biological analyses. However, current proteomic techniques, including two-dimensional polyacrylamide gel electrophoresis, multidimensional liquid chromatography systems, and gel and gel-free isoelectric focusing approaches such as chromatofocusing, immobilized pH membranes, Rotofor, free-flow electrophoresis, and off-gel electrophoresis, are all operated at the preparative-scale and are incompatible with small cell populations collected from microdissection-procured specimens. Our research goal is therefore to integrate the Gemini proteomic platform with tissue microdissection/preparation techniques as a novel biomarker discovery paradigm for enabling comprehensive and comparative survey of protein expression profiles in targeted cell populations isolated from clinical tissue specimens. The obvious progression in the development, evaluation, and validation of proposed biomarker discovery paradigm includes (i) the implementation of high throughput Gemini proteomic platform using a multiplexed nano-reversed-phase liquid chromatography system, (ii) full assessment of Calibrant's unique tissue proteome capabilities toward comparative characterization of protein expression profiles within microdissected tumor cells of glioma malignancies, and (iii) confirmation of discriminate biomarker candidates among different grades of glioma malignancies using Western blot and immunohistochemistry techniques on a large series of additional tissue specimens. In addition to the clinical proteomic technology development and validation, Phase II efforts will also focus on the identification of disease markers, exploration of molecular relationships among different disease states and phenotypes, and a deeper understanding of molecular mechanisms that drive disease progression. PUBLIC HEALTH RELEVANCE: Our research goal is to integrate the Gemini proteomic platform emerging from the Phase I project with tissue microdissection/preparation techniques as a novel biomarker discovery paradigm for enabling comprehensive and comparative survey of protein expression profiles in targeted cell populations isolated from clinical tissue specimens. The greatest expectations for targeted proteomic research using enriched malignant cells from high quality tissue specimens reside in the identification of diagnostic, prognostic, and predictive biological markers in the clinical setting, as well as the discovery and validation of new protein targets in the biopharmaceutical industry.

描述（由申请人提供）：仅当研究器官或组织样品由均质细胞群组成时，才能生成生物学和临床相关的蛋白质组学数据，其中没有不同类型和/或发育阶段的不需要细胞掩盖结果。分析组织样品的主要问题之一是样品的异质性质。组织活检中通常存在许多不同的细胞类型，在患病组织的情况下，少量的异常细胞可能位于或与未受影响的区域内或附近。因此，已经开发了激光捕获/显微解剖过程，以提供一种快速且直接的方法来采购生化和分子生物学分析的细胞或结构的均匀亚群。然而，当前的蛋白质组学技术，包括二维聚丙烯酰胺凝胶电泳，多维液相色谱系统，以及无凝胶和无凝胶的等电聚焦方法从显微解散的样品收集的种群。因此，我们的研究目标是将Gemini蛋白质组学平台与组织显微解剖/制备技术相结合，作为一种新型的生物标志物发现范式，以实现从临床组织分离的靶向细胞群体中蛋白质表达谱的全面和比较调查。提出的生物标志物发现范式的开发，评估和验证的明显进展包括（i）使用多重纳米相位液相色谱系统的高吞吐量Gemini蛋白质组学平台实施（iii）使用Western印迹和免疫组织化学技术在不同等级的神经胶质瘤恶性肿瘤中确认歧视生物标志物候选物，这是在大量其他组织标本上的。除了临床蛋白质组学技术的开发和验证外，第二阶段的工作还将集中在疾病标志物的鉴定，对不同疾病状态和表型之间的分子关系的探索以及对驱动疾病进展的分子机制的更深入了解。公共卫生相关性：我们的研究目标是将I阶段项目中出现的Gemini蛋白质组学平台与组织显微解剖/制备技术作为一种新型的生物标志物发现范式，以启用与临床组织标本中靶向细胞群体中蛋白质表达谱的全面和比较调查。使用高质量组织标本的富含恶性细胞的目标蛋白质组学研究的最大期望在于鉴定临床环境中诊断，预后和预测性生物学标志物，以及在生物药物行业中发现和验证新蛋白质靶标的新蛋白质。