Circulating tumor DNA in lymphoma: technologies and applications

Abstract

Lymphoma, a malignant tumor derived from lymphocytes and lymphoid tissues, presents with complex and heterogeneous clinical manifestations, requiring accurate patient classification for appropriate treatment. While invasive pathological examination of lymph nodes or lymphoid tissue remains the gold standard for lymphoma diagnosis, its utility is limited in cases of deep-seated tumors such as intraperitoneal and central nervous system lymphomas. In addition, biopsy procedures carry an inherent risk of complications. Computed tomography (CT) and positron emission tomography/computed tomography (PET/CT) imaging are essential for treatment assessment and monitoring, but lack the ability to detect early clonal evolution and minimal residual disease (MRD). Liquid biopsy-based analysis of circulating tumor DNA (ctDNA) offers a non-invasive alternative that allows for repeated sampling and overcomes the limitations of spatial heterogeneity and invasive biopsies. ctDNA provides genetic and epigenetic insights into lymphoma and serves as a dynamic, quantifiable biomarker for diagnosis, risk stratification, and treatment response. This review comprehensively summarizes common genetic variations in lymphoma and systematically evaluates ctDNA detection technologies, including PCR-based assays and next-generation sequencing (NGS). Applications of ctDNA detection in noninvasive genotyping, risk stratification, therapeutic response monitoring, and MRD detection are discussed across various lymphoma subtypes, including diffuse large B-cell lymphoma, Hodgkin lymphoma, follicular lymphoma, and T-cell lymphoma. By integrating recent research findings, the review highlights the role of ctDNA profiling in advancing precision medicine, enabling personalized therapeutic strategies, and improving clinical outcomes in lymphoma.

Keywords: Circulating tumor DNA; Liquid biopsy; Lymphoma; Minimal residue disease; Next-generation sequencing.

Fig. 1

Genetic features of ctDNA in lymphoma. Quantitative and qualitative analysis of ctDNA provide insights into cellular turnover, genetic and epigenetic alterations, immunoglobulin gene rearrangements, and fragmentomics, revealing underlying physiological and pathological processes in lymphoma

Fig. 2

Overview of lymphoma types, subtypes, genetic changes, and impacted signaling pathways in this review. Orange: lymphoma subtypes; light blue: the cell-of-origin classification of DLBCL; green: genetic subtypes of lymphoma; dark gray: genetic hallmarks of lymphoma; light gray: deregulated biological pathways. Abbreviations: DLBCL, diffuse large B-cell lymphoma; MCL, mantle cell lymphoma; PCNSL, primary central nervous system lymphoma; BL, Burkitt lymphoma; PTCL, peripheral T-cell lymphoma; FL, follicular lymphoma; cHL, classical Hodgkin lymphoma. N/A, not available

Fig. 3

Summary of key ctDNA detection technologies in lymphoma. Abbreviations: PCR, polymerase chain reaction; BEAMing, beads, emulsion, amplification and magnetics; ASO-PCR, allele specific oligonucleotide polymerase chain reaction; CNVs, copy number variants; ddPCR, droplet digital polymerase chain reaction; NGS, next generation sequencing; IgHTS, immunoglobulin high-throughput sequencing; VDJ, variable, diversity, joining; WGS, whole genome sequencing; WES, whole exome sequencing; CAPP-seq, cancer personalized profiling by deep sequencing; SNVs, single nucleotide variants; PhasED-seq, phased variant enrichment and detection sequencing; TBS, Targeted bisulfite sequencing

Fig. 4

Clinical applications of ctDNA testing in lymphoma. ctDNA profiling provides a noninvasive liquid biopsy method to capture comprehensive molecular characteristics, overcoming the limitations of traditional tissue sampling. Applications include diagnosis, genotyping, outcome prediction, response assessment, and MRD monitoring in lymphoma

Fig. 5

Key challenges in implementing ctDNA testing in clinical practice