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Observational Study
. 2025 Apr 8;9(7):1692-1701.
doi: 10.1182/bloodadvances.2024014136.

Molecular clustering on ctDNA improves the prognostic stratification of patients with DLBCL compared with ctDNA levels

Riccardo Moia  1 , Donatella Talotta  1 , Lodovico Terzi Di Bergamo  2 , Mohammad Almasri  1 , Riccardo Dondolin  1 , Matin Salehi  2 , Chiara Cosentino  1 , Roberta Soscia  3 , Irene Della Starza  3 , Alessio Bruscaggin  2 , Annalisa Andorno  4 , Francesca Mercalli  4 , Stefania Cresta  4 , Riccardo Bomben  5 , Tamara Bittolo  5 , Filippo Vit  5 , Pietro Bulian  5 , Antonella Zucchetto  5 , Riccardo Bruna  1 , Giulia Maria Rivolta  1 , Mattia Schipani  1 , Eleonora Secomandi  1 , Sreekar Kogila  1 , Matteo Bellia  1 , Samir Mouhssine  1 , Jana Nabki  1 , Bashar Al Deeban  1 , Joseph Ghanej  1 , Luca Cividini  1 , Nawar Maher  1 , Federica Melle  6 , Giovanna Motta  7 , Monica Leutner  8 , Angela Lorenzi  9 , Abdurraouf Mokhtar Mahmoud  1 , Wael Al Essa  1 , Clara Deambrogi  1 , Silvia Rasi  1 , Luigi Petrucci  3 , Renzo Luciano Boldorini  4 , Alice Di Rocco  3 , Ilaria Del Giudice  3 , Michele Spina  10 , Stefano Palazzolo  11 , Fabio Canal  12 , Vincenzo Canzonieri  11 , Maurizio Martelli  3 , Stefano Pileri  6 , Valter Gattei  5 , Robin Foà  3 , Davide Rossi  2   13   14 , Gianluca Gaidano  1
Affiliations
Observational Study

Molecular clustering on ctDNA improves the prognostic stratification of patients with DLBCL compared with ctDNA levels

Riccardo Moia et al. Blood Adv. .

Abstract

Circulating tumor DNA (ctDNA) levels can help predict outcomes in diffuse large B-cell lymphoma (DLBCL), but its integration with DLBCL molecular clusters remains unexplored. Using the LymphGen tool in 77 DLBCL cases with both ctDNA and tissue biopsy, a 95.8% concordance rate in molecular cluster assignment was observed, showing the reproducibility of molecular clustering on ctDNA. A multicenter, prospective cohort of 166 patients with newly diagnosed DLBCL was analyzed for ctDNA levels and molecular clusters using cancer personalized profiling by deep sequencing. Patients with ctDNA levels of <2.5 log10 haploid genome equivalents (hGE)/mL had a 4-year progression-free survival (PFS) and overall survival (OS) of 71.7% and 85.7%, respectively, compared with 50.3% and 61.0% for those with higher ctDNA levels (P = .0018 and P = .0017). Recursive partitioning showed that patients with ctDNA levels of ≥2.5 log10 hGE/mL were further stratified by clusters ST2/BN2. In this group, ST2/BN2 patients associated with a favorable outcome with a 4-year PFS and OS of 87.5% and 100%, respectively, compared to 38.0% and 47.1% for other clusters (P = .003 and P = .001). Combining ctDNA levels and ST2/BN2 clusters improved outcome prediction. Low-risk patients (n = 51), characterized by ctDNA levels of <2.5 log10 hGE/mL and/or BN2/ST2 cluster, had a 4-year PFS and OS of 75.3% and 87.8%, respectively. High-risk patients (n = 115), with ctDNA levels of ≥2.5 log10 hGE/mL and no BN2/ST2 cluster, had a 4-year PFS and OS of 38.0% and 47.1%, respectively. Adding cluster assignment to ctDNA levels improved the model's C statistics (0.63 vs 0.59 for PFS; 0.68 vs 0.63 for OS). Liquid biopsy thus provides a multilayered approach for outcome prediction in DLBCL.

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Conflict of interest statement

Conflict-of-interest disclosure: The authors declare no competing financial interests.

Figures

None
Graphical abstract
Figure 1.
Figure 1.
Molecular characterization of patients with DLBCL. (A) Case-level genomic profile of the 77 patients with DLBCL analyzed on ctDNA and on the tissue biopsy. Each column represents 1 patient and each row represents 1 genomic abnormality. Mutations identified on ctDNA are represented in gray, and mutations identified on the tissue biopsy are red. (B) Sankey plot showing the comparison of the assignment of each molecular cluster in the tissue biopsy and on ctDNA. (C) Case-level genomic profile of the 166 DLBCLs analyzed on ctDNA. Each column represents 1 patient and each row represents 1 genomic abnormality. Cell of origin (COO) and/or molecular clusters are plotted above each heat map. GC, germinal center; NA, not available.
Figure 1.
Figure 1.
Molecular characterization of patients with DLBCL. (A) Case-level genomic profile of the 77 patients with DLBCL analyzed on ctDNA and on the tissue biopsy. Each column represents 1 patient and each row represents 1 genomic abnormality. Mutations identified on ctDNA are represented in gray, and mutations identified on the tissue biopsy are red. (B) Sankey plot showing the comparison of the assignment of each molecular cluster in the tissue biopsy and on ctDNA. (C) Case-level genomic profile of the 166 DLBCLs analyzed on ctDNA. Each column represents 1 patient and each row represents 1 genomic abnormality. Cell of origin (COO) and/or molecular clusters are plotted above each heat map. GC, germinal center; NA, not available.
Figure 2.
Figure 2.
Prognostic impact of ctDNA levels and molecular clusters. Kaplan-Meier estimates of (A) PFS and (B) OS according to ctDNA levels. Patients with ctDNA levels of <2.5 log10 hGE/mL are represented by the blue curve and patients with ctDNA levels of ≥2.5 log10 hGE/mL are represented by the red curve. (C) Recursive partitioning plot according to ctDNA levels and assignment to clusters ST2/BN2. Kaplan-Meier estimates of (D) PFS and (E) OS in patients with ctDNA levels of ≥2.5 log10 hGE/mL according to ST2/BN2 molecular clusters. Patients assigned to ST2/BN2 are represented by the blue curve. Patients not assigned to ST2/BN2 are represented by the red curve.
Figure 3.
Figure 3.
Prognostic impact of ctDNA levels and ST2/BN2 molecular clusters. Kaplan-Meier estimates of (A) PFS and (B) OS according to ctDNA levels and ST2/BN2 clusters. Patients with ctDNA of <2.5 log10 hGE/mL and/or assigned to BN2/ST2 clusters are represented by the blue curve. Patients with ctDNA of ≥2.5 log10 hGE/mL and not assigned to cluster BN2/ST2 cluster are represented by the red curve. Multivariate analysis of (C) PFS and (D) OS including ctDNA levels, BN2/ST2 cluster, International Prognostic Index (IPI) score, and cell of origin.
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References

    1. Sehn LH, Salles G. Diffuse large B-cell lymphoma. N Engl J Med. 2021;384(9):842–858. - PMC - PubMed
    1. Tilly H, Morschhauser F, Sehn LH, et al. Polatuzumab vedotin in previously untreated diffuse large B-cell lymphoma. N Engl J Med. 2022;386(4):351–363. - PMC - PubMed
    1. Swerdlow SH, Campo E, Pileri SA, et al. The 2016 revision of the World Health Organization classification of lymphoid neoplasms. Blood. 2016;127(20):2375–2390. - PMC - PubMed
    1. Alizadeh AA, Eisen MB, Davis RE, et al. Distinct types of diffuse large B-cell lymphoma identified by gene expression profiling. Nature. 2000;403(6769):503–511. - PubMed
    1. Chapuy B, Stewart C, Dunford AJ, et al. Molecular subtypes of diffuse large B cell lymphoma are associated with distinct pathogenic mechanisms and outcomes. Nat Med. 2018;24(5):679–690. - PMC - PubMed