# Whole-exome sequencing is feasible on a fresh-frozen skin sample of intravascular large B cell lymphoma

## TL;DR

Researchers successfully used whole-exome sequencing on a skin sample from a rare lymphoma to identify key mutations, showing a feasible method for studying this challenging disease.

## Contribution

Demonstrates the feasibility of using fresh-frozen skin samples for whole-exome sequencing in intravascular large B-cell lymphoma.

## Key findings

- High-quality DNA was obtained from a fresh-frozen cutaneous sample of IVLBCL.
- Oncogenic mutations reflecting the post-germinal center origin of IVLBCL were identified.
- Restricting analysis to subcutaneous tissue improved cancer cell fraction and sequencing success.

## Abstract

Intravascular large B-cell lymphoma (IVLBCL) is a rare aggressive extranodal non-Hodgkin lymphoma. The predominant, if not exclusive, growth of neoplastic cells within the lumina of small-sized vessels represents the hallmark of the disease. Diagnosis is challenging due to the absence of marked lymphadenopathy, the highly heterogeneous clinical presentation, and the rarity of the condition. Clinical presentation is characterized by variable combinations of nonspecific signs and symptoms (such as fever and weight loss), organ-specific focal manifestations due to altered perfusion, and hemophagocytic syndrome. The rarity of this entity and the paucity of neoplastic cells in biopsy samples hamper the study of recurrent molecular abnormalities. The purpose of this study was to explore the feasibility of a different approach to recover a sufficient amount of DNA of acceptable quality to perform next-generation sequencing studies. Here, we report the findings of whole-exome next-generation sequencing performed on a fresh-frozen cutaneous sample of IVLBCL, paired with the patient saliva used as germline DNA. To increase the cancer cell fraction, only the subcutaneous tissue was selected. With this approach, we obtained high-quality DNA and were able to identify oncogenic mutations specific for this entity and recapitulating its post-germinal center origin, even if the tumor fraction was low. Molecular studies performed on fresh-frozen cutaneous sample are feasible in IVLBCL, especially when analysis is restricted to the subcutaneous tissue. Wide adoption of this reproducible and cost-effective approach may foster further studies, which may be of help in supporting diagnosis, providing pathogenetic insights, and guiding treatment decisions.

The online version contains supplementary material available at 10.1007/s10238-024-01308-0.