OVARIAN CANCER PROTEOME VIA TISSUE MICRODISSECTION AND GEMINI TECHNOLOGIES

通过组织显微切割和 Gemini 技术研究卵巢癌蛋白质组

基本信息

批准号：
7656860
负责人：
Satya Prakash Saxena
金额：
$ 26.03万
依托单位：
CALIBRANT BIOSYSTEMS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-11 至 2010-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7656860
关键词：
Adopted Archives Area Arizona Automobile Driving Back Behavior Benign Biochemical Biochemical Pathway Bioinformatics Biological Biological Markers Biological Markers Biological Products Biopsy Blood capillaries CA-125 Antigen Cancerous Carcinoma Cells Characteristics Classification Clinical Clinical Course of Disease Clinical Trials Collaborations Collection Complex Coupled Critical Pathways Cystadenoma DNA DNA copy number Data Detection Development Diagnosis Diagnostic Diagnostic Neoplasm Staging Disease Distant Metastasis Drug Approval Processes Electrophoresis Electrospray Ionization Environment Epidermal Growth Factor Receptor Epithelial Epithelium Evaluation Event Freezing Functional disorder Gel Genes Genetic Transcription Goals Gynecologic Gynecologic Oncology Group Gynecology Heterogeneity Histologic Human Human Genome Immunohistochemistry Industry Information Management Investigation Ions Isoelectric Focusing Label Lesion Liquid Chromatography Loss of Heterozygosity Malignant - descriptor Malignant Neoplasms Malignant neoplasm of ovary Manuals Mass Spectrum Analysis Measurement Medical Membrane Methods Microdissection Molecular Molecular Abnormality Molecular Profiling Monitor Neoplasm Metastasis Non-Malignant Normal Cell Organ Ovarian Ovarian Carcinoma Ovarian Serous Adenocarcinoma Ovarian Surface Epithelial-Stromal Tumor Ovarian Tissue Paraaortic Pathologist Pathology Patients Pelvic Examination Pelvis Performance Pharmaceutical Preparations Phase Plasma Play Polymorphism Analysis Population Post-Translational Protein Processing Preclinical Drug Evaluation Preclinical Testing Predictive Factor Preparation Process Protein Analysis Proteins Proteome Proteomics RNA Relative (related person)Reporting Reproducibility Research Role Sampling Serous Serum Serum Albumin Severities Shotguns Single Nucleotide Polymorphism Site Small Business Technology Transfer Research Specimen Staging Surface Surveys System TP53 gene Techniques Technology Testing Tissue Banks Tissue Sample Tissues Transvaginal Ultrasound Tumor Markers Two-Dimensional Polyacrylamide Gel Electrophoresis United States Food and Drug Administration Universities Validation Variant Western Blotting World Health Organization anticancer research base cDNA Arrays cancer cell capillary carcinogenesis clinically relevant comparative comparative genomic hybridization expectation human tissue improved laser capture microdissection lymph nodes medical schools neoplastic cell novel product development prognostic protein expression protein profiling public health relevance response success symposium therapeutic target tool tumor uncertain malignant potential neoplasm

项目摘要

DESCRIPTION (provided by applicant): Biologically and clinically relevant proteomics 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. Thus, several tissue microdissection technologies have been developed to provide a rapid and straightforward method for procuring homogeneous subpopulations of cells for biochemical and molecular biological analyses. However, current proteomic techniques, including 2-D PAGE, multidimensional liquid chromatography systems, and gel and gel-free isoelectric focusing approaches such as chromatofocusing, immobilized pH membranes, Rotofor, and free-flow electrophoresis, are all operated at the preparative-scale and are incompatible with small cell populations collected from microdissection-procured specimens. Thus, the reported tissue proteomics studies are mainly based on the analysis of entire tissue sections instead of targeted cell subpopulations. Clearly, development of the capability to enable tissue-based clinical proteomic studies will have far reaching impacts on protein biomarker investigations of disease through interrogation of the archived tissue collections. Our research goal is therefore to combine the novel tissue sample preparation and proteomic technologies that enable comprehensive and comparative survey of protein expression profiles in targeted tumor cell populations isolated from human tissues. By combining Calibrant's unique ability to perform proteomic profiling from minute samples with the expertise offered by Dr. Wenxin Zheng at the University of Arizona Medical College in cancer pathology, gynecology, and tissue microdissection, the proposed research represents a synergistic effort toward the evaluation and validation of a novel biomarker discovery paradigm for enabling the proteomic 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 ovarian carcinoma will be realized also through the collaboration with Dr. Zheng, a gynecologic pathologist, to provide access to a collection of primary ovarian epithelial tumor specimens. PUBLIC HEALTH RELEVANCE:Comprehensive and comparative proteomics studies of microdissected ovarian epithelial tumor specimens are proposed in this STTR Phase I project and are expected to provide significant details at the global level on the molecular mechanisms associated with ovarian epithelial carcinogenesis. Identification of differentially expressed proteins that are characteristic of a clearly defined disease state paves the way for defining the molecular and biochemical pathways by which normal cells progress to cancerous states in addition to nurturing discovery of biological markers and therapeutic targets for ovarian cancer. The greatest expectations for targeted proteomics research using enriched non-malignant or malignant ovarian 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）：只有当研究的器官或组织样本由同质细胞群组成时，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的细胞掩盖结果。因此，已经开发了几种组织显微切割技术，以提供一种快速而直接的方法来获取同质的细胞亚群以进行生化和分子生物学分析。然而，当前的蛋白质组学技术，包括 2-D PAGE、多维液相色谱系统以及凝胶和无凝胶等电聚焦方法（例如色谱聚焦、固定 pH 膜、Rotofor 和自由流动电泳），均以制备规模运行并且与从显微切割获得的样本中收集的小细胞群不相容。因此，报道的组织蛋白质组学研究主要基于整个组织切片的分析，而不是目标细胞亚群的分析。显然，基于组织的临床蛋白质组学研究能力的发展将对通过询问存档的组织集合来对疾病的蛋白质生物标志物研究产生深远的影响。因此，我们的研究目标是将新颖的组织样品制备和蛋白质组学技术结合起来，从而能够对从人体组织中分离的目标肿瘤细胞群中的蛋白质表达谱进行全面和比较的调查。通过将 Calibrant 从微小样本中进行蛋白质组分析的独特能力与亚利桑那大学医学院 Wenxin Cheng 博士在癌症病理学、妇科和组织显微切割方面提供的专业知识相结合，拟议的研究代表了评估和验证的协同努力一种新型生物标志物发现范式，可对癌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。通过与妇科病理学家郑博士的合作，还将实现将由此产生的生物标志物发现平台应用于研究与卵巢癌相关的分子机制，以提供原发性卵巢上皮肿瘤标本的收集。公共健康相关性：STTR I 期项目提出对显微解剖的卵巢上皮肿瘤标本进行全面和比较的蛋白质组学研究，预计将在全球水平上提供与卵巢上皮癌发生相关的分子机制的重要细节。鉴定作为明确定义的疾病状态特征的差异表达蛋白，除了促进卵巢癌生物标志物和治疗靶点的发现之外，还为定义正常细胞进展到癌状态的分子和生化途径铺平了道路。使用来自高质量组织标本的富集非恶性或恶性卵巢细胞进行靶向蛋白质组学研究的最大期望在于在临床环境中鉴定诊断、预后和预测生物标志物，以及发现和验证新的蛋白质靶标在生物制药行业。