Tissue Microarray Design and Development for Novel Gene

识别在肿瘤细胞中重要的基因和基因产物是国家癌症研究所的一个高度优先目标。为了确定感兴趣的基因是否在人类恶性肿瘤中表达，研究人员必须能够评估人类肿瘤的实际组织样本。不幸的是，获得大量的良好表征的人类组织样品是困难的，并且为了进行免疫组织化学或原位杂交，从肿瘤制备载玻片需要大量的费用。作为该问题的部分解决方案，病理学家开始将多个组织样本联合收割机组合在一个组织块中，以减少组织学成本以及进行特殊研究所涉及的时间和精力。这种技术已经被改进到可以将数百个组织样本以网格排列放置在单个石蜡组织块中的程度。NHGRI的研究人员（Kohenen，Kallioniemi等人）在《科学》杂志上发表的一篇开创性论文中证明了大规模组织微阵列的实用性。 1999年秋季NCI主办的组织微阵列研讨会之后，召开了一个指导委员会，目的是建立基于病理学的组织微阵列核心设施。指导委员会由来自校内和校外NCI以及NHGRI的代表组成。该项目是2001年旁路预算计划中确定的定义癌细胞特征的非凡机会的直接产物。第一年的目标是在NHGRI类似设施的基础上建立核心微阵列生产设施，并创建第一个全国分布的混合肿瘤微阵列。核心的初始产品是混合肿瘤块，含有最常见的上皮恶性肿瘤（乳腺、结肠、肺、前列腺和卵巢）的代表性样品以及黑色素瘤、神经胶质瘤和淋巴瘤的样品。还包括正常组织和标准细胞系的选择，以使组织点的总数达到500。通过CHTN获得了经分析的人类肿瘤样本，该组织还将处理将载玻片上的阵列分发给校内和校外科学界的研究人员。在第一个生产年度（2001财年）制备了3套5个多肿瘤阵列，总计近3000个载玻片用于分发。在过去的一年（2003财年），我们开发了3个阵列，以实现更集中的一个阵列。我们正在制作前列腺癌阵列、乳腺癌/卵巢癌阵列和肺癌/结肠癌阵列。我们还将继续制作更多样化的多肿瘤阵列，但前列腺样本将被肾癌、头颈部和甲状腺癌的切片所取代。除了制作载玻片外，核心设施还从事技术开发，从基本的组织学程序到阵列的使用，再到阵列的大规模成像。今年，我们收购了一台自动化点样仪，并在自动化点样仪和自动化成像系统的进一步技术开发上进行了合作。除了CHTN的肿瘤阵列外，我们还为NCI和NIH研究人员合作生产更小的定制阵列。

Identifying genes and gene products that are important in tumor cells is a high priority goal of the National Cancer Institute. In order to determine whether a gene of interest is expressed in human malignancy, investigators must be able to evaluate actual tissue samples of human tumors. Unfortunately, access to large numbers of well-characterized human tissue samples is difficult, and there is a large expense for the preparation of glass slides from tumor in order to perform immunohistochemistry or in-situ hybridization. As a partial solution to this problem, pathologists began to combine multiple tissue samples in one tissue block, in order to reduce the histology costs and the time and effort involved to perform the special studies. This technique has been refined to the point where hundreds of tissue samples can be placed in a grid arrangement in a single paraffin tissue block. Investigators in the NHGRI (Kohenen, Kallioniemi and others) demonstrated the utility of large scale tissue microarrays in a seminal paper published in Science. Following a tissue microarray workshop hosted by NCI in Fall, 1999, a steering committee convened with the purpose of establishing a pathology-based tissue microarray core facility. The steering committee is composed of representatives from intramural and extramural NCI and from the NHGRI. This project is a direct outgrowth of the Extraordinary Opportunity to Define the Signatures of Cancer Cells identified in the 2001 Bypass Budget plan. The goals of the first year were to establish a core microarray production facility based on a similar facility in the NHGRI and to create the first mixed tumor microarrays for nationwide distribution. The initial product of the core was a mixed tumor block containing representative samples of the most common epithelial malignancies (breast, colon, lung, prostate, and ovary) as well as samples of melanoma, glioma and lymphoma. A selection of normal tissue and standard cell lines was also included to bring the total number of tissue spots to 500. Anonymized human tumor samples are obtained through the CHTN, and this organization will also handle distribution of arrays on glass slides to investigators in the intramural and extramural scientific community. Three sets of 5 multi-tumor arrays were prepared in the first production year (FY2001), totaling nearly 3000 slides for distribution. This past year (FY2003) we have developed 3 arrays for more focused one. We are making a prostate carcinoma array, a breast/ovarian carcinoma array and a lung/colon carcinoma array. We will also continue to produce a more diverse multitumor array, but the prostate samples will be replaced by sections with renal carcinoma, head and neck and thyroid carcinomas. In addition to slide production, the core facility has been engaged in technology development, from the basic histology procedures to array use to large-scale imaging of arrays. This year we acquired an automated arrayer and have worked collaboratively on further technology development of the automated arrayer and an automated imaging system. In addition to the tumor arrays for the CHTN, we produce smaller custom arrays for NCI and NIH investigators on a collaborative basis.