Development of FISH Molecular Diagnostics to support Clinical Trials at NCI

染色体病理单元（CPU）于2010年开放，以满足“内部”实验室FISH测试的需求，以支持NCI的临床试验。CPU团队已于2010年11月22日完成了官方CAP认证和CLIA认证。从2011年1月起，实验室开始接收临床病例进行FISH检测。中央处理器自启用以来进行了5000多次测试。这包括超过1120个临床报告测试的FISH分析，超过5000个探针验证测试以及与LP和其他NIH研究人员的合作。FISH诊断是病理学实验室目前提供的病理学分子诊断的一个组成部分。在最近的CAP报告中发现，FISH诊断的敏感性和特异性接近100%，这使其在其他基于PCR和免疫组织化学的可用测试中处于领先地位，这些测试长期存在已知的特异性问题。病理实验室的染色体病理单元（CPU）对福尔马林固定石蜡包埋（FFPE）组织进行荧光原位杂交（FISH）分析，以支持NCI和NIH的积极临床试验。对这种测试的需求很高；CPU每天接受临床肿瘤样本FISH测试的订单，目前支持NCI的17项临床试验。这包括来自整个CCR研究人员社区的请求：分子肿瘤科、外科科、主任办公室、儿科肿瘤科、神经肿瘤科、泌尿肿瘤科。活性检测菜单包括FISH检测，用于实体瘤和血液恶性肿瘤中临床显著的染色体易位和扩增事件。开发/验证的检测包括Her-2在乳腺癌、肺癌和胃肠道癌中的扩增，c-myc在Burkitt和弥漫性大B细胞淋巴瘤中的易位，BCL2在滤泡性淋巴瘤中的易位，脑肿瘤中的1p/19q缺失检测，肺癌和炎性肌纤维母细胞瘤中的ALK易位，肺癌中的FGFR1、PDGFRA和PIK3A扩增，胸腺肿瘤中的NTRK1扩增，肾肿瘤中的TFE-3易位，肺泡软组织肉瘤，EWSR1/FLI1和foxo1易位在儿童肿瘤（尤文氏肉瘤、Rabdomyosarcoma）等中的作用此外，CPU目前正在开发和验证十种新的FISH测试，用于NCI研究人员要求的癌症中其他常见的染色体异常。正在进行的试验包括甲状腺癌的RET重排、前列腺癌的ERG易位、肾肿瘤的TFE-B基因易位（占儿童肾癌的30%）。CPU一直致力于开发新的诊断方法，这将大大推进目前癌症临床测试的现有方法。其中之一是从癌症患者外周血中分离和分子分析循环肿瘤细胞（CTCs），作为检测和监测肿瘤转移性疾病和治疗效率的一种非侵入性活检方法。该单位配备了最先进的实验室，其中包括来自BioView的新型自动荧光成像工作站。蔡司（Zeiss）的apooome显微镜，具有允许FISH分析和厚组织切片成像的apooome功能，SKY-cube用于光谱核型-用于检测癌细胞和各种遗传综合征的染色体易位的现代工具。

The Chromosome Pathology Unit (CPU)was opened in 2010 to fulfill the need in the "in house" laboratory FISH tests required to support clinical trials at NCI. The official CAP accreditation and CLIA certification has been accomplished by the CPU team in November 22, 2010. From January 2011 the Lab started receiving clinical cases for FISH testing. The CPU performed more than 5,000 tests since its opening. This includes FISH assays for over 1120 clinical reported tests, over 5000 tests for probe validation and collaboration with LP and other NIH researchers. FISH diagnostics are an integral part of the Pathology molecular diagnostics currently offered by the Lab of Pathology. The sensitivity and specificity of FISH diagnostics was found to be near 100% in recent reports of CAP that puts them in front line among other available tests based on PCR and immunohistochemistry that have a long-known specificity problem. The Chromosome Pathology Unit (CPU) in the Laboratory of Pathology performs Fluorescence In Situ Hybridization (FISH) assays on formalin-fixed paraffin-embedded (FFPE) tissues to support active clinical trials at the NCI and NIH. The demand for this testing is high; the CPU receives orders for FISH tests on clinical tumor samples daily, and currently supports 17 clinical trials at the NCI. That includes requests from the entire community of CCR investigators: Molecular Oncology Branch, Surgery Branch, Office of the Director, Pediatric Oncology Branch, Neuro-Oncology Branch, Urologic Oncology Branch. The active test menu includes FISH assays for clinically significant chromosomal translocations and amplification events in solid tumors and hematologic malignancies. The developed/ validated tests include Her-2 amplification in Breast, Lung, and GI tract cancer, c-myc translocation in Burkitt and Diffuse Large B Cell Lymphoma, BCL2 translocation in Follicular Lymphoma, 1p/19q deletion tests for brain tumors, ALK translocation in lung cancer and inflammatory myofibroblastic tumors, FGFR1, PDGFRA and PIK3A amplification in lung cancer, NTRK1 amplification in thymic tumors, TFE-3 translocation in renal tumors, alveolar soft part sarcoma, EWSR1/FLI1 and FOXO1translocations in pediatric tumors (Ewing's sarcoma and Rabdomyosarcoma), etc. In addition, the CPU is currently developing and validating ten new FISH tests for additional frequent chromosomal abnormalities in cancers that have been requested by NCI investigators. Among the tests in the pipeline are RET rearrangements in thyroid cancer, ERG translocation in prostate cancer, TFE-B gene translocation in renal tumors (responsible for 30% of pediatric kidney cancer). The CPU has been working on developing novel diagnostics that would significantly advance currently existing approaches in cancer clinical testing. One of them is Isolation and Molecular Profiling of the Circulating Tumor Cells (CTCs) from the peripheral blood of cancer patients as an alternative non-invasive biopsy method for detection and monitoring tumor metastatic disease and treatment efficiency. The Unit has been equipped as state-of-the art laboratory that includes a new automated Fluorescence imaging workstation Duet from BioView. Inc, Zeiss epifluorescence microscope with the ApoTome feature that allows FISH analysis and imaging of thick tissue sections, SKY-cube for spectral karyotyping - a modern tool for the detection of chromosomal translocations in cancer cells and various hereditary syndromes.