Patient-derived lymphoma spheroids reveal predictive markers of glofitamab resistance in relapsed/refractory B-NHL
Abstract
Abstract Bispecific antibodies (bsAbs) such as glofitamab represent a promising therapeutic approach for relapsed/refractory B-cell non-Hodgkin lymphoma (R/R B-NHL), but resistance mechanisms remain poorly understood. This study aimed to identify predictive markers of bsAb resistance based on the response of 3-dimensional patient-derived lymphoma spheroids (PDLS) established from 39 R/R B-NHL samples. PDLS were treated with glofitamab for 3 days, and B-cell depletion was quantified to assess the ex vivo treatment response. Comprehensive immune profiling was performed on patient samples using multiparametric flow cytometry, single-cell RNA sequencing, codetection by indexing spatial proteomics, and functional assays. High responders to glofitamab possessed CD8+ T cells with consistently higher cytotoxic and activation signatures across effector differentiation states, whereas low responders showed enrichment of exhausted CD8+ T cells with enhanced expression of exhaustion markers (T-cell immunoglobulin and ITIM domain [TIGIT], LAG3, and PD1). Furthermore, low responders exhibited elevated functional CD4+ T follicular helper (Tfh) cells in close proximity to malignant B cells, thus promoting their survival through interleukin-21 and C-X-C motif chemokine ligand 13 signaling pathways. Analysis of pretreatment RNA-sequencing data from 48 patients with R/R B-NHL confirmed that high Tfh cell abundance is associated with poor glofitamab response. In PDLS, anti-TIGIT cotreatment enhanced glofitamab efficacy in low responders, and Tfh cell depletion experiments confirmed that reducing Tfh cell activity increased B-cell depletion. Together, these findings identify CD8+ T-cell exhaustion and functionally activated Tfh cells as key factors associated with glofitamab resistance in R/R B-NHL. This work supports their potential use as predictive biomarkers for selecting patients with higher probability of response and provides a foundation for future combination therapeutic strategies.
Article Details
Authors (25)
Paul Marcoux
1Université de Toulouse, INSERM, Centre de Recherches en Cancerologie de Toulouse, Toulouse, France
Fabien Gava
1Université de Toulouse, INSERM, Centre de Recherches en Cancerologie de Toulouse, Toulouse, France
Marie Tosolini
Pauline Gravelle
Christina Schniederjohann
6Department of Hematology, Oncology and Clinical Immunology, Medical Faculty and University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, Germany
Sonia Quertinmont
1Université de Toulouse, INSERM, Centre de Recherches en Cancerologie de Toulouse, Toulouse, France
Neus Serrat
11Fundació Clínic per a la Recerca Biomèdica, Institut d'Investigacions Biomèdiques August Pi i Sunyer, Barcelona, Spain
Fanny Bouquet
13F. Hoffmann-La Roche, Basel, Switzerland
Sylvia Herter
Karin Tarte
15Institut National de la Santé et de la Recherche Médicale, Unité Mixte de Recherche U127, Université Rennes, Etablissement Français du Sang Bretagne, LabEx IGO, Rennes, France
Mikael Roussel
15Institut National de la Santé et de la Recherche Médicale, Unité Mixte de Recherche U127, Université Rennes, Etablissement Français du Sang Bretagne, LabEx IGO, Rennes, France
Pierre Sesques
17Department of Hematology, University Hospital of Lyon, Lyon, France
Caroline Bret
18Institut de Génétique Humaine, Centre Hospitalier Universitaire, Montpellier, France
Cédric Rossi
19Clinical Hematology, Dijon University Hospital, Dijon, France
Pierre Aubert
20Department of Hematology, Grenoble Alpes, Centre Hospitalier Universitaire Grenoble Alpes, Grenoble, France
Franck Morschhauser
Centre Hospitalier Universitaire de Lille, Groupe de Recherche sur les formes Injectables et les Technologies Associées, Lille, France
Guillaume Cartron
CHU Montpellier UMR5535, Montpellier, France
Wolfgang Huber
Sascha Dietrich
Loic Ysebaert
13Service Hématologie, Institut Universitaire du Cancer de Toulouse-Oncopole, Toulouse, France
Pierre Brousset
Peter-Martin Bruch
6Department of Hematology, Oncology and Clinical Immunology, Medical Faculty and University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, Germany
Patricia Pérez-Galán
Christine Bezombes
1Université de Toulouse, INSERM, Centre de Recherches en Cancerologie de Toulouse, Toulouse, France
Camille Laurent