OVARIAN CANCER PROTEOME VIA TISSUE MICRODISSECTION AND GEMINI TECHNOLOGIES

描述（由申请人提供）：生物学和临床相关的蛋白质组学数据只能在被调查的器官或组织样本由均匀的细胞群组成，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的情况下产生。因此，已经开发了几种组织显微解剖技术，以提供一种快速和直接的方法来获取均匀的细胞亚群，用于生化和分子生物学分析。然而，目前的蛋白质组学技术，包括二维PAGE、多维液相色谱系统、凝胶和无凝胶等电聚焦方法，如色谱聚焦、固定化pH膜、Rotofor和自由流电泳，都是在制备规模上操作的，与从微解剖获得的标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，而不是靶向细胞亚群。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对存档组织收集的询问来研究疾病的蛋白质生物标志物产生深远的影响。因此，我们的研究目标是将新的组织样品制备和蛋白质组学技术相结合，从而能够对从人体组织中分离的靶向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在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、妇科和组织显微解剖方面的专业知识相结合，该提议的研究代表了对一种新的生物标志物发现范式的评估和验证的协同努力，该范式能够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。将由此产生的生物标志物发现平台应用于卵巢癌相关分子机制的研究也将通过与妇科病理学家郑博士的合作来实现，以提供对原发性卵巢上皮肿瘤标本的收集。公共卫生相关性：在这个STTR I期项目中，提出了对微解剖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。鉴别具有明确定义的疾病状态特征的差异表达蛋白，为确定正常细胞向癌症状态发展的分子和生化途径铺平了道路，此外还有助于发现卵巢癌的生物标志物和治疗靶点。利用高质量组织标本中富集的非恶性或恶性卵巢细胞进行靶向蛋白质组学研究，最大的期望在于临床环境中诊断、预后和预测生物标志物的鉴定，以及生物制药行业中新蛋白质靶点的发现和验证。

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.