Cytologic and Molecular Diagnostics for Vitreoretinal Lymphoma: Current Approaches and Emerging Single-Cell Analyses

Abstract

Vitreoretinal lymphoma (VRL) is a rare ocular malignancy that manifests as diffuse large B-cell lymphoma. Early and accurate diagnosis is essential to prevent mistreatment and to reduce the high morbidity and mortality associated with VRL. The disease can be diagnosed using various methods, including cytology, immunohistochemistry, cytokine analysis, flow cytometry, and molecular analysis of bulk vitreous aspirates. Despite these options, VRL diagnosis remains challenging, as samples are often confounded by low cellularity, the presence of debris and non-target immunoreactive cells, and poor cytological preservation. As such, VRL diagnostic accuracy is limited by both false-positive and false-negative outcomes. Missed or inappropriate diagnosis may cause delays in treatment, which can have life-threatening consequences for patients with VRL. In this review, we summarize current knowledge and the diagnostic modalities used for VRL diagnosis. We also highlight several emerging molecular techniques, including high-resolution single cell-based analyses, which may enable more comprehensive and precise VRL diagnoses.

Circulating cell-free DNA and IL-10 from cerebrospinal fluids aid primary vitreoretinal lymphoma diagnosis

“…However, the false negative rate of vitreous biopsy cytology or immune cytology is approximately 70% ( 8 , 9 ). Flow cytometry of vitreous fluid increases the diagnostic sensitivity, up to 82% ( 10 , 11 ).…”

Refractory Primary Vitreoretinal Lymphoma Involving the Spinal Cord with a Temporary Complete Response to Tirabrutinib

“…To address these unmet needs, several emerging molecular techniques, including high-resolution single-cell-based analyses and gene expression profiling analyses, have been developed. These techniques can identify actionable genomic alterations from small-volume, intraocular liquid biopsies and determine the biological characteristics of PVRL (11,12). MYD88 mutations, especially L265P, are very frequent in PVRL, and their detection allows a definitive diagnosis of lymphoma to be made, even with poor-quality samples (13).…”

Primary Vitreoretinal Lymphoma: Current Diagnostic Laboratory Tests and New Emerging Molecular Tools

“…Despite being considered a valuable tool for VRL detection, with elevated sensitivity (82.4%) and specificity (100%) [ 41 ], it has to be taken into account that flow cytometry requires a large number of viable cells for diagnosis [ 37 , 42 ].…”

“…Hence, the low number of intact neoplastic cells in vitreous fluid, which is often combined with the presence of numerous inflammatory/reactive cells, unfortunately represents a critical limitation for the use of this technique in PVRL diagnosis [ 37 , 42 ].…”