Molecular Diagnostics Core Laboratory

分子诊断核心实验室

• 批准号: 9154319
• 负责人: Mark Raffeld
• 金额: $ 63.51万
• 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/9154319
• 关键词: Alleles; American; Autoimmune Diseases; Azathioprine; B-Lymphocytes; BCL1 Oncogene; BCL2 gene; BRAF gene; Behavior; Biological Assay; Biological Markers; Capillary Electrophoresis; Classification; Clinical; Clinical Trials; Clonality; Collaborations; Cutaneous; Cutaneous Melanoma; DNA; Detection; Development; Diagnosis; Diagnostic; Disease; ERBB2 gene; EWS-FLI1 fusion protein; Early Diagnosis; Eligibility Determination; Epidermal Growth Factor Receptor; Future; Genes; Genotype; Glioblastoma; Glioma; Human Herpesvirus 4; Human Herpesvirus 8; Immunotherapy; Inflammatory Bowel Diseases; KRAS2 gene; Laboratories; Laboratory Research; Link; Lymphoma; Lymphoproliferative Disorders; MGMT gene; Malignant Neoplasms; Malignant neoplasm of brain; Malignant neoplasm of lung; Methods; Methylation; Molecular; Molecular Diagnostic Testing; Molecular Target; Mutation; Mutation Analysis; National Heart, Lung, and Blood Institute; National Human Genome Research Institute; National Institute of Allergy and Infectious Disease; PIK3CA gene; Pathology; Patients; Pediatric Neoplasm; Plasma; Process; Prognostic Marker; Proteins; Proto-Oncogene Proteins c-akt; Recurrence; Reporting; Research; Research Personnel; Resources; Reverse Transcriptase Polymerase Chain Reaction; STAT3 gene; Sampling; Satellite Viruses; Structure; T-Cell Lymphoma; T-Lymphocyte; Technology; Testing; Transcript; Translational Research; United States National Institutes of Health; Viral Load result; Yang; austin; base; bisulfite; childhood sarcoma; circulating DNA; clinically relevant; college; epidermal growth factor receptor VIII; gene panel; improved; melanoma; molecular oncology; neoplastic cell; next generation sequencing; novel; prognostic; programs; promoter; pyrosequencing; research clinical testing; response; targeted treatment; temozolomide; therapeutic target; tumor; tumor DNA

The Molecular Diagnostics Laboratory is currently the only CLIA and College of American Pathology approved clinical laboratory within the NCI certified for performing molecular oncology testing on pathology materials from NIH patients. In 2014 the molecular diagnostics laboratory processed 1774 unique clinical samples from NCI/NIH patients, and performed 6742 tests. The laboratory utilizes a variety of technologies to perform these assays including conventional DNA PCR, RT-PCR, qPCR, allele suppression PCR, capillary electrophoresis, pyrosequencing, bisulfite pyrosequencing, and small panel next generation sequencing (NGS). The assays we perform include tests to identify B and T-cell clonality, lymphoma specific translocations (e.g., BCL-1/IG, BCL-2/IG), translocations associated with pediatric sarcomas (e.g., EWS-FLI-1, EWS-ERG, EWS-WT-1, PAX-7/FKHD, PAX-3/FKHD, SYT-SSX-1, and SYT-SSX-2), cancer associated viruses (e.g., EBV, HTLV, HHV-8), and mutations associated with a variety of cancers (e.g., EGFR, KRAS, NRAS, BRAF, PIK3CA, IDH1/2, AKT, MYD88, ERBB2, STAT3, and STAT5b). Quantitative PCR is performed for HTLV 1/2 to follow viral load in ATL clinical trials, and for the EGFRvIII to assess expression of this cancer-specific transcript for clinical trial eligibility. MGMT methylation analysis is performed using bisulfite pyrosequencing to provide predictive information regarding response to temozolomide in the glioblastoma setting. IDH1/2 is performed to assist in glioma diagnosis and as a prognostic marker. The laboratory also offers a 50 gene NGS panel for clinical use, interrogating some of the more commonly involved genes in cancer. This panel is currently our standard CLIA level mutation panel when it is necessary to assess mutations in more than one gene as is the case in lung cancer. The laboratory's developmental research effort has been focused on the application of next generation sequencing (NGS) technologies to assist in cancer genotyping and diagnostics. In addition to the 50 gene panel currently in clinical use, the laboratory is developing more focused tumor specific panels that will better cover the mutational spectra of specific tumor types. The laboratory has also initiated a program in the use of circulating tumor DNA (ctDNA) as a biomarker for disease response and early detection. These two efforts (NGS and ctDNA) are linked as the genotyping is not only used to identify potential therapeutic targets, but it is also used to identify targets for the circulating DNA studies. Early studies in melanoma patients (collaboration with Dr. S. Rosenberg) indicate the potential use of this technology in predicting responses to immunotherapy and recurrence, and we are now extending these studies in both the cutaneous cutaneous melanoma setting and in other cancers, initiating collaborations with other NCI PIs. We see these efforts as having great potential to move into clinical use here at the NCI, and elsewhere, in the near future. Among our more notable laboratory initiated studies reported in 2014 was the identification of STAT3 and STAT5b mutations in gamma delta hepatosplenic T-cell lymphoma, a rare lymphoma subtype associated with patients with autoimmune disease (especially inflammatory bowel disease) treated with azathioprine. This is the first report of a specific mutation being associated with this rare lymphoma. The molecular diagnostics laboratory also supports translational research of NCI and NIH researchers. Among the NCI and NIH investigators and clinicians that have utilized the laboratory's resources in 2014 are: Dr. A. Apolo (NCI), Dr. Fred Barr (NCI), Dr. Austin Barrett (NHLBI), Dr. William Gahl (NHGRI), Dr. Steven Holland (NIAID), Dr. Elaine Jaffe (NCI), Dr. Udai Kammula (NCI) Dr. Daniel Kastner (NHGRI) Dr. Amy Klion (NHLBI) Dr. Robert Kreitman (NCI) Dr. Markku Mietinnen (NCI), Dr. Kenneth Olivier (NIAID), Dr. Stefania Pittaluga (NCI), Dr. Martha Quezado (NCI), Dr. Koneti Rao (NIAID) ,Dr. Steven Rosenberg (NCI), Dr. Helen Su (NHLBI), Dr. Jeffrey Cohen (NIAID), Dr. Thomas Uldrick (NCI), Dr. Gulbu Uzel (NIAID), Dr. Katherine Warren (NCI), Dr. Wyndham Wilson (NCI), Dr. James Yang (NCI), Dr. Neil Young (NHLBI).

该分子诊断实验室目前是NCI认证的唯一CLIA和美国病理学学院，该实验室在NCI认证的临床实验室中，用于对NIH患者的病理材料进行分子肿瘤测试。 2014年，分子诊断实验室处理了NCI/NIH患者的1774个独特的临床样本，并进行了6742次测试。实验室利用各种技术来执行这些测定法，包括常规的DNA PCR，RT-PCR，QPCR，等位基因抑制PCR，毛细管电泳，Pyrosequencing，Bisulfite Pyrosequencing和小型面板下一代测序（NGS）。 The assays we perform include tests to identify B and T-cell clonality, lymphoma specific translocations (e.g., BCL-1/IG, BCL-2/IG), translocations associated with pediatric sarcomas (e.g., EWS-FLI-1, EWS-ERG, EWS-WT-1, PAX-7/FKHD, PAX-3/FKHD, SYT-SSX-1, and SYT-SSX-2），癌症相关病毒（例如EBV，HTLV，HHV-8）以及与多种癌症相关的突变（例如EGFR，KRAS，NRAS，NRAS，BRAF，PIK3CA，IDH1/2，IDH1/2，AKT，AKT，MYD88，ERBB2，STAT3，STAT3，STAT3和Stat5B）。对HTLV 1/2进行定量PCR，以在ATL临床试验中跟随病毒载量，并为EGFRVIII评估该癌症特异性转录本以表达临床试验资格。使用Bisulfite Pyrosequencing进行MGMT甲基化分析，以提供有关胶质母细胞瘤环境中对替莫唑胺的反应的预测信息。进行IDH1/2以协助神经胶质瘤诊断和预后标记。该实验室还提供了一个50个基因NGS面板供临床使用，询问了一些更常见的癌症基因。当需要像肺癌那样评估多个基因的突变时，该小组是我们的标准CLIA水平突变面板。实验室的发展研究工作集中在下一代测序（NGS）技术方面的应用，以帮助癌症基因分型和诊断。除了目前用于临床用途的50个基因面板外，实验室还开发了更集中的肿瘤特定面板，可以更好地覆盖特定肿瘤类型的突变光谱。该实验室还启动了一项计划，用于使用循环肿瘤DNA（CTDNA）作为疾病反应和早期检测的生物标志物。这两项努力（NGS和CTDNA）是连接的，因为基因分型不仅用于识别潜在的治疗靶标，而且还用于识别循环DNA研究的靶标。对黑色素瘤患者的早期研究（与S. Rosenberg博士的合作）表明，该技术在预测对免疫疗法和复发的反应中的潜在用途，我们现在在皮肤皮肤黑色素瘤的环境和其他癌症中扩展了这些研究，并在其他癌症中启动了与其他NCI PIS的合作。我们认为，这些努力具有在不久的将来在NCI和其他地方进入临床用途的巨大潜力。在2014年报道的我们更著名的实验室发起研究中，是发现与甲酸酯治疗的自身免疫性疾病（尤其是炎性肠道疾病）相关的罕见的淋巴瘤亚型STAT3和STAT5B突变，这是一种罕见的淋巴瘤亚型。这是与这种罕见淋巴瘤相关的特定突变的第一个报告。分子诊断实验室还支持NCI和NIH研究人员的转化研究。在2014年利用实验室资源的NCI和NIH调查人员和临床医生中，A。Apolo博士（NCI），Fred Barr博士（NCI），Austin Barrett博士（NHLBI）博士（NHLBI）博士William Gahl（NHGRI）博士，NHGRI（NHGRI），Steven Holland（Niaid）博士（NIAID（NIAID）博士，Elaine Jaffe（nci niel jaffe） Kastner（NHGRI）Amy Klion博士（NHLBI）Robert Kreitman博士（NCI）Markku Mietinnen博士（NCI），Kenneth Olivier博士（NIAID），Stefania Pittaluga博士（NCI），Martha Quezado（NCI） Steven Rosenberg（NCI），Helen Su（NHLBI）博士，Jeffrey Cohen博士（Niaid）博士，Thomas Uldrick博士（NCI），Gulbu Uzel（Niaid）博士，Katherine Warren（NCI）博士，NCI（NCI），Wyndham Wilson（NCI），Wyndham Wilson（NCI），詹姆斯·Yang（NCI）。