Figure 4.

Longitudinal information from AH liquid biopsy for Case 33. Fundus photos, tumor fraction (TFx), cfDNA quantification, estimated clinical tumor volume from B-scan measurements, and genomic profiles for each clinical timepoint at which an AH sample was evaluated. This eye was non-responsive to treatment, and ultimately required secondary enucleation (ES) due to an apical tumor recurrence with persistently active seeds. A dramatic decrease in cfDNA quantity was observed over time, consistent with the decrease in the main tumor volume. This indicates that the total number of tumor cells that were actively shedding cfDNA into the AH decreased. However, TFx remained at a high level throughout treatment, reflecting the persistent seeding that still contributes to tumor-derived cfDNA in the AH. Without much physiological cfDNA to dilute the tumor-derived cfDNA, the TFx remained high. Genomic profiles consistently demonstrated the same 3 SCNAs that were present at diagnosis, as well as a new large-scale 2p gain (*) and 19q loss in the AH of the enucleated eye, suggesting clonal evolution at the time of recurrence.