Importance: Uveal Melanomas (UM) micro-metastasis can be present prior to diagnosis and relapse after treatment. Earlier detection resulted in an increased incidence of small (T1 and T2) tumors allowing for novel eye-preserving treatment strategies but reducing available tumor tissue needed for prognostic genomic profiling, creating the need for minimal-invasive detection and novel prognostication methods. Objective: To determine whether tumor presence can be confirmed using metabolite patterns in blood plasma and to evaluate if these patterns differ between high risk (BRCA1-associated protein-1, BAP1), intermediate risk (Splicing Factor 3b Subunit 1, SF3B1) and low risk (Eukaryotic Translation Initiation Factor 1A X-Linked, EIF1AX) mutated tumors. Design: Retrospective observational study including discovery (n=53) and replication (n=42) convenience sample sets compared to unaffected control-participants (n=46) as well as across mutation-based subgroups. Setting: Patients from two tertiary referral centers specialized in ocular oncology: The Rotterdam Eye Hospital and the Erasmus MC Cancer Institute were included. Participants: Sex-matched controls and patients were included based on their prognostic relevant secondary driver mutations. Peripheral blood plasma was collected at diagnosis, prior to treatment. Exclusion criteria were the presence of other malignancies or co-occurrence of systemic diseases at time of diagnosis. Main outcome and measure: Metabolite profiles of patients and control-participants were generated as mass/charge (m/z) features using ultra-high performance liquid chromatography mass-spectrometry. After normalization, discriminatory feature patterns were determined using a random forest classifier and leave-one-out cross-validation. Results: We detected differential metabolic patterns with a sensitivity of 0.95 and 0.90 and a specificity of 0.98 and 0.98 in the positive and negative ion modes, respectively. The accuracy of the model for classifying the subgroups was insufficient for the discovery (0.600 and 0.614 in the positive and negative ion modes, respectively) and replication cohort (0.544 and 0.672 in the positive and negative ion modes, respectively). Conclusion and relevance: Minimally invasive metabolomics does not discriminate between the prognostic relevant BAP1, SF3B1 and EIF1AX mutated UM-subgroups. However, this technique has the potential to allow for minimal invasive screening as it distinguishes metabolite patterns in peripheral blood derived plasma of UM-patients from control-participants.

The authors have declared no competing interest.

This study was funded by CORR (Combined Ophthalmic Research Rotterdam) and the BAYER Ophthalmology research award. The Combined Ophthalmic Research Rotterdam Biobank of the Department of Ophthalmology and the Rotterdam Eye Hospital is supported by the Combined Ophthalmic Research Rotterdam Foundation.

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Medical Ethics Committee of Erasmus MC gave ethical approval for this work

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All data produced in the present study are available upon reasonable request to the authors