Tissue Microarray Design for Novel Gene/Product Screenin

鉴定肿瘤细胞中重要的基因和基因产物是国家癌症研究所的首要目标。 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，获得大量特征明确的人体组织样本很困难，并且从肿瘤制备载玻片以进行免疫组织化学或原位杂交需要花费大量费用。 作为这一问题的部分解决方案，病理学家尝试将多个组织样本组合在一个组织块中，然后可以用来减少组织学成本以及进行特殊研究所需的时间和精力。 这项技术已经被改进到可以将数百个组织样本以网格排列方式放置在单个石蜡组织块中。 NHGRI 的研究人员（Kohenen、Kallioniemi 等）在《科学》杂志上发表的一篇开创性论文中展示了大规模组织微阵列的实用性。 1999 年秋季，NCI 主办了一次组织微阵列研讨会后，召开了一个指导委员会，旨在建立基于病理学的组织微阵列核心设施。 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 的类似设施的基础上建立核心微阵列生产设施，并创建第一个在全国范围内销售的混合肿瘤微阵列。 核心设施的空间被分配在先进技术核心中，并聘请了一名组织技术专家来处理建立组织学实验室的技术问题以及进行块切割和染色活动。 由来自院内和院外 NCI 以及 NHGRI 的代表组成的指导委员会确定，核心的第一个产品应该是混合肿瘤块，其中包含最常见的上皮恶性肿瘤（乳腺、结肠、肺、前列腺和卵巢）的代表性样本以及黑色素瘤、神经胶质瘤和淋巴瘤的样本。 还包括精选的正常组织和标准细胞系，使组织斑点总数达到约 700 个。肿瘤样本是通过 CHTN 获得的，该组织还将负责将载玻片上的阵列分发给校内和校外科学界的研究人员。 第一个多肿瘤阵列的当前目标日期是 2000 年 11 月，阵列载玻片将分发用于 beta 测试。 然后，指导委员会将决定核心将准备的未来肿瘤块，但选项包括罕见肿瘤（肾细胞、肝细胞、肉瘤）的特殊块、从与预后相关的集合（例如乳腺癌的 NSABP 集合）准备的块或特定于特定临床试验的块。

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 expence 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 have tried to combine multiple tissue samples in one tissue block, that could then be used 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. 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. Space for the core facility was assigned in the Advanced Technology Core and a histotechnologist was hired to handle the technical aspects of establishing the histology laboratory and to perform block cutting and staining activities. The steering committee, which is composed of representatives from intramural and extramural NCI and from the NHGRI, determined that the first product of the core should be 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 were also to be included to bring the total number of tissue spots to approximately 700. Tumor samples were 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. The current target date for the first multitumor arrays is November, 2000 and arrayed slides will be distributed for beta testing. The steering committee will then make decisions about future tumor blocks that the core will prepare, but options include both specialty blocks of rarer tumors (renal cell, hepatocellular, sarcomas), blocks prepared from collections linked to prognosis (e.g. the NSABP collection of breast cancers) or blocks specific to particular clinical trials.