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 2013 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. In the last year, the laboratory developed and began to offer for clinical use a 50 gene NGS panel 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 the last year 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 2013 are: Dr. A. Apolo (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. Kenneth Olivier (NIAID) Dr. Stefania Pittaluga (NCI) Dr. Martha Quezado (NCI) Dr. Koneti Rao (NIAID) Dr. Steven Rosenberg (NCI) Dr. Helen Su (NHLBI) Dr. Thomas Uldrick (NCI) Dr. Gulbu Uzel (NIAID) Dr. Katherine Warren (NCI) Dr. Wyndham Wilson (NCI) Dr. James Yang (NCI) Dr. Neil Young (NHLBI).
Showing the most recent 10 out of 93 publications
