Development of FISH Molecular Diagnostics to support Clinical Trials at NCI

染色体病理学部门（CPU）于2010年成立，以满足支持NCI临床试验所需的“内部”实验室FISH检测的需求。CPU团队于2010年11月22日完成了官方CAP认证和CLIA认证。从2011年1月开始，实验室开始接收FISH检测的临床病例。自启用以来，CPU进行了5，000多次测试。这包括超过1120个临床报告测试的FISH检测，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诊断的灵敏度和特异性接近100%，这使得它们在基于PCR和免疫组织化学的其他可用测试中处于第一线，这些测试具有长期已知的特异性问题。病理学实验室的染色体病理学单元（CPU）对福尔马林固定石蜡包埋（FFPE）组织进行荧光原位杂交（FISH）检测，以支持NCI和NIH的积极临床试验。对这项检测的需求很高; CPU每天都会收到对临床肿瘤样本进行FISH检测的订单，目前支持NCI的17项临床试验。这包括CCR研究者整个社区的请求：分子肿瘤学分支、外科分支、主任办公室、儿科肿瘤学分支、神经肿瘤学分支、泌尿肿瘤学分支。活动测试菜单包括实体瘤和恶性血液病中具有临床意义的染色体易位和扩增事件的FISH检测。开发/验证的检测包括乳腺癌、肺癌和胃肠道癌中的Her-2扩增，伯基特和弥漫性大B细胞淋巴瘤中的c-myc易位，滤泡性淋巴瘤中的BCL 2易位，脑肿瘤的1 p/19 q缺失检测，肺癌和炎性肌纤维母细胞肿瘤中的ALK易位，肺癌中的FGFR 1、PDGFRA和PIK 3A扩增，胸腺肿瘤中的NTRK 1扩增，此外，中央政策组目前正在开发和验证10种新的FISH检测方法，用于NCI研究人员要求的癌症中其他常见的染色体异常。正在进行的测试包括甲状腺癌的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，这将大大推进癌症临床测试中现有的方法。其中之一是从癌症患者的外周血中分离和分子分析循环肿瘤细胞（CTC），作为检测和监测肿瘤转移性疾病和治疗效率的替代非侵入性活检方法。该单位已配备了最先进的实验室，包括一个新的自动化荧光成像工作站Duet从BioView。公司，蔡司落射荧光显微镜与ApoTome功能，允许FISH分析和成像的厚组织切片，SKY-cube光谱核型分析-一个现代化的工具，用于检测染色体易位的癌细胞和各种遗传综合征。

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.