Chemical cytometry for neoplasia prognosis-Barrett's esophagus

描述(申请人提供)：本申请是对RFA-CA-06-003《癌症研究新兴技术的应用》的响应。本RFA邀请研究项目的申请，以评估新兴分子技术的有用性，这些技术已准备好用于癌症研究的临床或生物学问题。项目应设计为证明该技术是可靠的，并产生可重复性的测量结果。项目还应设计为收集初步数据，以支持该技术在未来以临床或生物学为重点的项目(S)中的使用。该应用程序结合了华盛顿大学化学系和病理学系以及弗雷德·哈钦森癌症研究中心的研究人员的努力。这些研究人员将进行一项初步研究，以评估癌症研究的新技术--化学细胞术。这项技术监测单个细胞中蛋白质和生物胺的表达，并将用于从西雅图巴雷特食道队列患者的活检中分离出的细胞的研究。我们将系统地确定Barrett‘s食道活检组织中细胞成分的差异。我们将从从活检中分离的单个隐窝中解剖20个细胞，并对这些细胞进行化学细胞分析。这项研究将提供对隐窝和干细胞发育成其分化形式过程中蛋白质表达的差异的理解。我们将对从一次活检中获得的三个隐窝中分离的细胞进行化学细胞学分析。这些隐窝将是密切相关的，这项研究将提供关于克隆内变异的信息。我们将对从同一患者的三次活检中分离出的隐窝细胞进行化学细胞分析。本研究将提供有关克隆间变异的信息。我们将生成5名患者的指纹，这将提供关于患者之间差异的信息。作为这些研究的结果，我们将确定必要的细胞或隐窝的数量，以确定肿瘤的特征。基于这些结果，我们将提出一项后续拨款，用于系统验证化学细胞术对腺癌进展的预后，并开发适合大规模临床分析的平台。

DESCRIPTION (provided by applicant): This application is in response to RFA-CA-06-003, Applications of emerging technologies for Cancer Research. This RFA:" invites applications for research projects to evaluate the usefulness of emerging molecular technologies that are ready for initial application to clinical or biological questions in cancer research. Projects should be designed to demonstrate that the technology is robust and yields reproducible measurements. Projects should also be designed to gather preliminary data to support the use of the technology in a future project(s) with a clinical or biological focus." The application combines the efforts of researchers in the Departments of Chemistry and Pathology at the University of Washington and at the Fred Hutchinson Cancer Research Center. These researchers will perform a pilot study to evaluate new technology for cancer research, chemical cytometry. This technology monitors protein and biogenic amine expression in single cells, and will be evaluated for the study of cells isolated from biopsies obtained from patients in the Seattle Barrett's Esophagus Cohort. We will systematically determine the variance in cellular composition within Barrett's esophagus biopsies We will dissect 20 cells from a single crypt isolated from a biopsy and perform chemical cytometry on those cells. This study will provide understanding of the variance in protein expression along a crypt and during development of a stem cell into its differentiated form. We will perform chemical cytometry on cells isolated from three crypts obtained from a single biopsy. These crypts will be closely related and the study will provide information on the intraclonal variation. We will perform chemical cytometry on cells isolated from crypts obtained from three biopsies from the same patient. This study will provide information on the interclonal variation. We will generate fingerprints from 5 patients, which will provide information on the between-patient variance. As a result of these studies, we will have identified the number of cells or crypts necessary to be assayed to characterize the neoplasia. Based on these results, we will propose a follow-up grant for the systematic validation of chemical cytometry for prognosis of the progression to adenocarcinoma, and for the development of a platform appropriate for large-scale clinical analysis.