CITP-Based Selective Tissue Proteome Enrichment

基于 CITP 的选择性组织蛋白质组富集

基本信息

批准号：
7942540
负责人：
CHENG S LEE
金额：
$ 19.58万
依托单位：
UNIV OF MARYLAND, COLLEGE PARK
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-01 至 2013-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7942540
关键词：
Area Astrocytoma Biochemical Biological Biological Markers Blood capillaries Categories Cells Clinical Complex Consumption Coupling Data Databases Detection Development Diagnosis Disadvantaged Disease Dissection Electrophoresis Electrospray Ionization Environment Evaluation Figs - dietary Fractionation Freezing Glioblastoma Injection of therapeutic agent Label Liquid Chromatography Liquid substance Mass Fragmentography Measurement Methods Microdissection Microinjections Molecular Molecular Profiling National Institute of Neurological Disorders and Stroke Non-Receptor Tyrosine Kinase Gene Pathology Pathway interactions Performance Phase Phenotype Plasma Population Procedures Process Proteins Proteome Proteomics Pump Relative (related person)Reproducibility Research Saliva Sampling Site Specimen Speed Staging Staining method Stains Stroke Structure System Techniques Technology Tissue Model Tissue Sample Tissues Urine Validation Variant anticancer research base capillary comparative interest laser capture microdissection nano neoplastic cell nervous system disorder neuropathology novel protein expression public health relevance tumor tumor progression wasting

项目摘要

DESCRIPTION (provided by applicant): In contrast to universally enriching all analytes by a similar degree, the result of the capillary isotachophoresis (CITP) stacking process is that major components may be diluted, but trace compounds are concentrated. Such selective enhancement toward low abundance proteins will drastically reduce the range of relative protein abundances within complex tissue proteomes, and greatly enhance the proteome coverage using the CITP-based proteomic technology. Our proposed research efforts therefore aim to fully characterize, develop, and exploit the use of this differential concentration effect to achieve comprehensive proteome analysis of clinical specimens with limited sample availability. The laser capture microdissection (LCM) process provides a rapid and straightforward method for isolating selected subpopulations of cells for downstream biochemical and molecular analyses. On the basis of cell enrichment, LCM also serves as a targeted sample fractionation approach toward the reduction in protein complexity and relative abundance. The proposed coupling of tissue microdissection for diseased cell enrichment with CITP-based selective analyte concentration not only presents a synergistic strategy for the detection and characterization of low abundance proteins, but also offers a novel biomarker discovery paradigm for enabling the identification of tumor-associated markers, exploration of molecular relationships among different tumor states and phenotypes, and a deeper understanding of molecular mechanisms that drive cancer progression. PUBLIC HEALTH RELEVANCE: By combining our unique bioanalytical capabilities with the expertise of Dr. Zhengping Zhuang at the National Institute of Neurological Disorders and Stroke (NINDS) in tissue microdissection and neuropathology, the proposed research represents a synergistic effort toward the development, evaluation and validation of a novel biomarker discovery paradigm for enabling the proteomic analysis of tumor cells and their micro-environment in support of cancer research, diagnosis, and treatment.

描述（申请人提供）：与以相似程度普遍富集所有分析物相比，毛细管等速电泳（CITP）堆积过程的结果是主要成分可能被稀释，但痕量化合物被浓缩。这种针对低丰度蛋白质的选择性增强将大大减少复杂组织蛋白质组内相对蛋白质丰度的范围，并使用基于 CITP 的蛋白质组技术大大提高蛋白质组覆盖率。因此，我们提出的研究工作旨在充分表征、开发和利用这种差异浓度效应，以在可用样本有限的情况下实现临床样本的全面蛋白质组分析。激光捕获显微切割 (LCM) 过程提供了一种快速、直接的方法来分离选定的细胞亚群以进行下游生化和分子分析。在细胞富集的基础上，LCM 还可以作为一种有针对性的样品分级分离方法，以降低蛋白质的复杂性和相对丰度。所提出的将用于病变细胞富集的组织显微切割与基于 CITP 的选择性分析物浓缩相结合，不仅为低丰度蛋白质的检测和表征提供了协同策略，而且还提供了一种新的生物标志物发现范式，用于识别肿瘤相关标志物，探索不同肿瘤状态和表型之间的分子关系，并更深入地了解驱动癌症进展的分子机制。公共健康相关性：通过将我们独特的生物分析能力与国家神经疾病和中风研究所 (NINDS) 的庄正平博士在组织显微切割和神经病理学方面的专业知识相结合，拟议的研究代表了对开发、评估和验证的协同努力一种新型生物标志物发现范例，可对肿瘤细胞及其微环境进行蛋白质组学分析，以支持癌症研究、诊断和治疗。