OVARIAN CANCER PROTEOME VIA TISSUE MICRODISSECTION AND GEMINI TECHNOLOGIES

描述（由申请方提供）：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蛋白质组学数据，其中没有不同类型和/或发育阶段的多余细胞会掩盖结果。因此，已经开发了几种组织显微切割技术，以提供一种快速和直接的方法，用于获得用于生物化学和分子生物学分析的同质细胞亚群。然而，目前的蛋白质组学技术，包括2-D PAGE，多维液相色谱系统，凝胶和无凝胶等电聚焦方法，如chromatofocusing，固定化pH膜，Rotofor，和自由流动电泳，都是在电泳规模和不兼容的小细胞群体收集从显微解剖采购标本。因此，所报道的组织蛋白质组学研究主要基于对整个组织切片而不是靶细胞亚群的分析。显然，开发能够进行基于组织的临床蛋白质组学研究的能力将通过询问存档的组织集合对疾病的蛋白质生物标志物研究产生深远的影响。因此，我们的研究目标是联合收割机结合新的组织样品制备和蛋白质组学技术，使全面和比较调查的蛋白质表达谱在靶向肿瘤细胞群体分离自人类组织。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郑文新博士在癌症病理学、妇科学和组织显微切割方面的专业知识相结合，这项研究代表了一种协同努力，旨在评估和验证一种新的生物标志物发现范例，用于对癌细胞及其微生物进行蛋白质组学分析。支持癌症研究，诊断和治疗的环境。通过与妇科病理学家郑博士的合作，将所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，以提供对原发性卵巢上皮性肿瘤标本的收集。公共卫生相关性：在STTR I期项目中提出了对显微切割的卵巢上皮性肿瘤标本进行全面和比较蛋白质组学研究，并有望在全球水平上提供与卵巢上皮性癌发生相关的分子机制的重要细节。鉴别明确定义的疾病状态特征的差异表达蛋白质为定义正常细胞发展为癌性状态的分子和生化途径铺平了道路，此外还培育了卵巢癌生物标志物和治疗靶点的发现。使用来自高质量组织标本的富集的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，以及在生物制药行业中发现和验证新的蛋白质靶点。

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.