Tissue Print Micropeels for Molecular Profiling Cancer

描述(由申请人提供)：分子图谱已成为一种重要的策略，用于识别与表征特定癌症的生物学变化相关的标记“签名”。为了充分发挥丰富的新生物标志物信息的潜力，有必要开发出可在临床环境中使用的人类组织和肿瘤标本的分析策略。临床标本是异质性的，组织异质性是在应用分子图谱分析人类癌症时必须解决的主要复杂性来源之一。我们已经开发了一套新颖的“组织印迹”技术，使我们能够在不损害标本的情况下，在人体组织和肿瘤样本的大范围内描绘分子标记。我们首次将我们的新组织印迹技术应用于前列腺癌根治术标本中与包膜侵袭相关的蛋白质标记物的分析。最近，我们发现，在组织印迹收集过程中，我们可以从标本上剥离一层细胞，这个过程不会造成可检测到的组织损伤(由外科病理学家确定)，因此不会干扰常规的临床外科病理。我们还表明，在组织印迹“Micropeel”中收集的细胞足以进行聚合酶链式反应和定量RT-PCR分析，使我们能够对多个分子标记进行评分，并将结果组合成与样本表面相对应的“平铺图案”。 在本计划中概述的项目中，我们将与启元公司的研发团队密切合作，以优化从我们从人的前列腺和乳腺组织/肿瘤标本中收集的组织印迹微皮中的mRNA和DNA的产量。然后，我们将开发用于前列腺针活检的组织印迹微皮采样技术的原则验证试点应用，这是必须完整保存以进行临床诊断的一类标本。我们的长期目标是在临床环境中利用组织印迹技术来简化在活检和手术标本中获得足够的人类癌症代表性的过程，并为这个组织采样平台开发协议，以支持蛋白质组分析和基于聚合酶链式反应的DNA和mRNA分析技术。除了促进基础研究和转译研究外，组织打印平台还可以用作专门临床应用的工具，当标记物特征本身具有潜在的诊断重要性时，提供标本扩展区域的“分子切片”。

DESCRIPTION (provided by applicant): Molecular profiling has emerged as an important strategy for identifying marker "signatures" associated with the biological changes that characterize specific cancers. To realize the full potential of the wealth of new biomarker information, it is essential to develop strategies for profiling human tissue and tumor specimens that are workable in a clinical setting. Clinical specimens are heterogeneous, and tissue heterogeneity is one of the major sources of complexity that must be addressed in the application of molecular profiling to the analysis of human cancers. We have developed a set of novel "tissue print" techniques that allow us to profile the molecular markers over extended areas of human tissue and tumor samples without damaging the specimen. We first applied our new tissue print techniques in the profiling of protein markers associated with capsular invasion in radical prostatectomy specimens. More recently, we have discovered that, during tissue print collection, we can peel a layer of cells off of the specimen and that this process does not cause detectable tissue damage (as determined by surgical pathologists), and thus does not interfere with routine clinical surgical pathology. We have also shown that the cells collected in the tissue print "micropeel" are adequate for PCR and quantitative rt-PCR analysis, allowing us to score multiple molecular markers and assemble the results in "tiling patterns" corresponding to the specimen surface. In the project outlined in this proposal, we will work closely with the research and development team at Qiagen Inc. to optimize the yield of mRNA and DNA from our tissue print micropeels collected from human prostate and breast tissue/tumors specimens. We will then develop a proof-of-principle pilot application of the tissue-print micropeel sampling technique for prostate needle biopsies, one of the classes of specimens that must be conserved intact for clinical diagnosis. Our long term goal is to utilize tissue print techniques in the clinical setting to simplify the process of obtaining an adequate representation of human cancers in biopsies and surgical specimens, and to develop protocols for this tissue sampling platform to support both proteomic analysis and PCR-based DNA and mRNA profiling techniques. In addition to facilitating basic and translational research, the tissue-printing platform can also be utilized as a tool for dedicated clinical applications, to provide "molecular sections" of extended areas of the specimen when the marker profile is itself of potential diagnostic importance.