Ultradeep sequencing detects GNAQ and GNA11 mutations in cell‐free DNA from plasma of patients with uveal melanoma. (14th February 2013)

• Record Type: Journal Article
• Authors: Metz, Claudia H. D.; Scheulen, Max; Bornfeld, Norbert; Lohmann, Dietmar; Zeschnigk, Michael
• Is Part Of: Cancer medicine. Volume 2:Number 2(2013:Apr.)
• Page Start: 208
• Page End: 215
• Publication Date: 2013-02-14
• Subjects: 616.994005
• DOI: 10.1002/cam4.61
• Languages: English
• ISSNs: 2045-7634

Abstract

<abstract abstract-type="main" id="cam461-abs-0001"> <title>Abstract</title> <p>Elevated levels of cell‐free DNA (cfDNA) are frequently observed in tumor patients. Activating mutations in exon 4 (R183) and exon 5 (Q209) of <italic>GNAQ</italic> and <italic>GNA</italic><italic>11</italic> are almost exclusively found in uveal melanoma, thus providing a highly specific marker for the presence of circulating tumor DNA (ctDNA). To establish a reliable, noninvasive assay that might allow early detection and monitoring of metastatic disease, we determined the proportion of <italic>GNAQ</italic> or <italic>GNA</italic><italic>11</italic> mutant reads in cfDNA of uveal melanoma patients by ultradeep sequencing. Cell‐free DNA from 28 uveal melanoma patients with metastases or extraocular growth was isolated and quantified by real‐time polymerase chain reaction (PCR) (7–1550 ng DNA/mL plasma). <italic>GNAQ</italic> and <italic>GNA</italic><italic>11</italic> regions of interest were amplified in 22 of 28 patients and ultradeep sequencing of amplicons was performed to detect even low proportions of mutant reads. We detected Q209 mutations (2–38% mutant reads) in either <italic>GNAQ</italic> or <italic>GNA</italic><italic>11</italic> in the plasma of 9 of 22 metastasized patients. No correlation between the proportion of mutant reads and the concentration of cfDNA could be detected. Among the nine ctDNA‐positive patients, four had metastases in bone, whereas no metastases were detected in the 13 ctDNA‐negative patients at this location (<italic>P</italic> = 0.025). Furthermore, ctDNA‐positive patients tended to be younger at initial diagnosis and show larger metastases. The results show that ultradeep amplicon sequencing can be used to detect tumor DNA in plasma of metastasized uveal melanoma patients. It remains to be shown if this approach can be used for early detection of disseminated tumor disease.</p> </abstract>