Development of FISH Molecular Diagnostics to support Clinical Trials at NCI

开发 FISH 分子诊断以支持 NCI 的临床试验

• 批准号: 8554181
• 负责人: Svetlana Pack
• 金额: $ 52.39万
• 依托单位: DIVISION OF BASIC 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/8554181
• 关键词: 19q; Accreditation; Alveolar Soft Part Sarcoma; BCL2 gene; Biological Assay; Brain Neoplasms; Breast; CCR; Cancer Patient; Carbon Dioxide; Cells; Certification; Chromosomal Duplication; Chromosomal translocation; Chromosome abnormality; Chromosomes; Clinical; Clinical Trials; Collaborations; Communities; DNA; DNA Sequence Rearrangement; Data; Decision Making; Detection; Development; Diagnostic; ERBB2 gene; EWSR1 gene; Effectiveness; Epidermal Growth Factor Receptor; Equipment; Event; Ewings sarcoma; FGFR1 gene; FLI1 gene; FOXO1A gene; Fluorescent in Situ Hybridization; Follicular Lymphoma; Formalin; Gastrointestinal tract structure; Genes; Goals; Hematologic Neoplasms; Housing; Image Analysis; Immunohistochemistry; Immunotherapy; In Situ; In Situ Hybridization; Incubators; Inflammatory Pseudotumor; Inherited; Kidney Neoplasms; Laboratories; Lung; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of prostate; Measures; Medicine; Microscope; Molecular; NTRK1 gene; Operative Surgical Procedures; Outcome; PDGFRB gene; Paraffin Embedding; Pathology; Patients; Pediatric Neoplasm; Pediatric Oncology; Protocols documentation; Reporting; Research; Research Personnel; Roche brand of trastuzumab; Running; Sampling; Sensitivity and Specificity; Solid Neoplasm; Specificity; Spectral Karyotyping; Survival Rate; Syndrome; Testing; Thick; Thymus Neoplasms; Tissues; United States National Institutes of Health; Urologic Oncology; Validation; alternative treatment; base; c-Myc Staining Method; cancer cell; clinically significant; improved; large cell Diffuse non-Hodgkin' s lymphoma; malignant breast neoplasm; molecular oncology; molecular pathology; neuro-oncology; tool; tumor

The Chromosome Pathology Unit (CPU)was set up 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 2,600 tests since its opening. This includes FISH assays for over 860 clinical reported tests, 1740 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, cmyc 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, PDGFR and PIK3A amplification in lung cancer, NTRK1 amplification in thymic tumors, TFE-3 translocation in renal tumors, alveolar soft part sarcoma, EWSR1/FLI1 and FKHR translocations 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 FGFR1, ROS-1 and RET rearrangements in lung cancer, ERG translocation in prostate cancer, TFE-B gene translocation in renal tumors. The Unit has been equipped as state-of-the art laboratory that includes a new 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. All small equipment to measure the DNA quantity, run PCR to validate the FISH data, the CO2 incubator to grow cells were purchased as well.

染色体病理学部门（CPU）成立于2010年，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2，600多次测试。这包括超过860个临床报告测试的FISH检测，1740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cmyc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PDGFR和PIK 3A扩增，胸腺肿瘤中的NTRK 1扩增，TFE-3易位在肾肿瘤，腺泡状软组织肉瘤，EWSR 1/FLI 1和FKHR易位在儿科肿瘤（尤文氏肉瘤和横纹肌肉瘤）等，此外，中央处理局目前正在开发和验证10个新的FISH测试，额外的常见的染色体异常的癌症，已要求由NCI研究人员。正在进行的测试包括肺癌中的FGFR 1，ROS-1和RET重排，前列腺癌中的ERG易位，肾肿瘤中的TFE-B基因易位。该单位已配备了最先进的实验室，包括一个新的蔡司落射荧光显微镜与ApoTome功能，允许荧光原位杂交分析和成像的厚组织切片，天空立方体光谱核型分析-一个现代化的工具，检测染色体易位癌细胞和各种遗传综合征。还购买了用于测量DNA量、运行PCR以验证FISH数据、用于培养细胞的CO2培养箱的所有小型设备。