Prospective implementation of an aqueous humor liquid biopsy platform informs clinical diagnosis and management of retinoblastoma and other intraocular lesions

ABSTRACT

LBSeq4Kids is a clinically validated liquid biopsy platform combining low passage whole genome sequencing (LP-WGS) for copy number alterations (CNAs) and a custom cancer targeted sequencing panel (TSP) to detect sequence variants in cell-free DNA from the aqueous humor (AH) of the eye, cerebrospinal fluid, and plasma. We present LBSeq4Kids results from a prospective cohort of 60 ocular oncology patients, including 41 with retinoblastoma (RB),13 with non-malignant RB simulating lesions and six with other intraocular malignancies. Ninety-four percent of baseline RB samples obtained at diagnosis were positive for CNAs by LP-WGS and 83% were positive for pathogenic variants by TSP analysis. All samples obtained at clinical recurrence were positive for ctDNA whereas none of the eyes in remission had a positive finding. The presence of CNAs detected by serial sampling in patients being treated for RB was correlated with clinical disease status. None of the patients with RB-simulating lesions had a positive finding by LP-WGS. The sensitivity of the assay to detect ctDNA in the setting of active RB was 98%. LBSeq4Kids represents a groundbreaking improvement for intraocular malignancies and is highly effective in informing accurate diagnosis, risk stratification, response to therapy, and surveillance.