Tissue Microarray Design and Development for Novel Gene

新基因的组织微阵列设计和开发

• 批准号: 7331339
• 负责人: DAVID E KLEINER
• 金额: --
• 依托单位: DIVISION OF CLINICAL SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7331339
• 关键词: Tissue; Microarray; Design; Development; Novel

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 pathology-based tissue microarray core facility was established. 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. Four sets of multi-tumor arrays were prepared in the first production year (FY2001), totaling more than 3000 slides for distribution. In the current phase of this project we are collaborating with mainly intramural investigators on tumor specific projects, both building arrays as well as staining and analyzing the pathology. This past year (FY2005) we have built arrays of the NCI60 cell lines and xenografts from those cell lines, pediatric tumors, breast carcinoma, non-small cell lung carcinoma, melanoma and general tumor arrays. 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. We have acquired an automated arrayer and have worked collaboratively on further technology development of the automated arrayer and an automated imaging system.

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，获得大量特征良好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大量费用。作为这一问题的部分解决方案，病理学家开始将多个组织样本合并在一个组织块中，以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。继 1999 年秋季由 NCI 主办的组织微阵列研讨会之后，建立了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 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。核心的初始产品是混合肿瘤块，包含最常见的上皮恶性肿瘤（乳腺癌、结肠癌、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 500 个。匿名人类肿瘤样本是通过 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。在第一个生产年（2001 财年）就制备了四套多肿瘤芯片，总共分发了 3000 多张载玻片。在该项目的当前阶段，我们主要与校内研究人员合作开展肿瘤特定项目，包括构建阵列以及染色和分析病理学。去年（2005 财年），我们构建了 NCI60 细胞系和这些细胞系的异种移植物、儿科肿瘤、乳腺癌、非小细胞肺癌、黑色素瘤和普通肿瘤阵列的阵列。除了玻片生产外，核心设施还从事技术开发，从基本组织学程序到阵列使用，再到阵列的大规模成像。我们购买了一台自动阵列仪，并合作开发自动阵列仪和自动成像系统的进一步技术。