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Sarah Lake, Bertil Damato, Helen Kalirai, Lisa Olohan, Xuan Liu, Sarah Coupland; Identification of regulators of uveal melanoma metastasis through whole exome sequencing. Invest. Ophthalmol. Vis. Sci. 2013;54(15):4530.
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© ARVO (1962-2015); The Authors (2016-present)
The drivers of uveal melanoma (UM) metastasis are not yet fully understood despite studies identifying a potential function for genes including BAP1, MDA-9, DDEF1 and ID2. This study aimed to identify genes that are mutated in primary, metastasising, UM and which may therefore be additional, key regulators of UM metastasis.
DNA from 12 clinically and histologically well-defined UM was sequenced following exome capture (50Mb SureSelect, Agilent) using the SOLiD system (Applied Biosystems). Tumours were stratified according to their risk of developing metastasis using an accelerated failure time model and included six monosomy 3 UM with high-risk of metastasis, and six disomy 3 UM with low metastatic risk.1 A systems biology approach (GeneGo, Metacore) was used to identify mutated genes that occurred in all high-risk UM and which were therefore likely to influence metastatic cell behaviour. Bidirectional, chain-termination, sequencing was used to confirm the presence of mutations. Expression of the encoded proteins was assessed by immunohistochemistry.
Seven genes had non-synonymous mutations in their coding regions. These were: LAMB2 (chr 3), USP19 (chr 3), ULBP3 (chr 6), ARFGEF1 (chr 8), EPHA1 (chr 7), FOS (chr 14) and KIF23 (chr 15). A statistically-significant difference in the frequency of mutations in the high-risk UM compared to the low-risk UM was observed (p<0.003-0.046). These mutations were not present in dbSNP (http://www.ncbi.nlm.nih.gov/projects/SNP/). No mutations of BAP1 were identified in any UM. Copy number alterations and/or insertion/deletions of genes were not consistently seen in the high-risk UM. Protein expression of the mutated genes was detected in UM, regardless of metastatic risk, for USP19, ULBP3, ARFGEF1, EPHA1 and KIF23. No expression of FOS or LAMB2 protein was detected in UMs.
Investigations are underway to determine whether the mutations detected in the coding regions of USP19, ULBP3, ARFGEF1, EPHA1 and KIF23 alter the biological function of the encoded proteins to promote metastatic progression. References 1. Eleuteri A DB, Coupland SE and Taktak AFG. Enhancing survival prognostication in patients with choroidal melanoma by integrating pathologic, clinical and genetic predictors of metastasis. Int J Biomedical Engineering and Technology 2012;8:18-35.
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