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333 CD70 Deregulation in Follicular Lymphoma at Diagnosis Is Associated with Relapse and Opens New Avenues for Dual CD19-CD70 CAR-T Therapy

Program: Oral and Poster Abstracts
Type: Oral
Session: 622. Lymphomas: Translational – Non-Genetic: Novel Immune Evasion Strategies in B Cell Lymphomas
Hematology Disease Topics & Pathways:
Research, Fundamental Science, Translational Research, Lymphomas, Non-Hodgkin lymphoma, B Cell lymphoma, Chimeric Antigen Receptor (CAR)-T Cell Therapies, Diseases, Indolent lymphoma, Immune mechanism, Cell expansion, Treatment Considerations, Biological therapies, Lymphoid Malignancies, Biological Processes, Technology and Procedures, Gene editing
Saturday, December 7, 2024: 4:30 PM

Ferran Araujo-Ayala1,2*, Maria Ros2,3*, Mireia Bachiller, PhD4*, Raluca Alexandru5*, Pablo Mozas, MD, PhD6*, Salut Colell7*, Judith Mateos-Jaimez8*, Cristina López, PhD2,7*, María Villalba-Esparza, PhD5*, Ferran Nadeu, PhD7*, Andrea Rivero, MD9*, Cèlia Dobaño-López, PhD8*, Juan Garcia Valero, PhD2,7*, Neus Serrat, MD, PhD10*, Marta Gimenez-Alejandre, PhD7*, Ariadna Giró7*, Daniel J Hodson, MD-PhD11, Armando Lopez-Guillermo, MD6,12, Elias Campo, MD, PhD13, Dolors Colomer14,15*, Alba Maiques-Diaz7*, Jose Ignacio Martin-Subero, PhD7,16*, Silvia Bea7,14,17,18*, Laura Magnano, MD, PhD19*, Sonia Guedan20*, Carlos Eduardo De Andrea, MD, PhD5* and Patricia Perez Galan, PhD2,8*

1Fundació de Recerca Clínic Barcelona - Institut d’Investigacions Biomèdiques August Pi I Sunyer (FRCB-IDIBAPS), Barcelona, ESP
2Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), Madrid, Spain
3Fundació de Recerca Clínic Barcelona - Institut d’Investigacions Biomèdiques August Pi I Sunyer (IDIBAPS), Barcelona, Spain
4Fundació de Recerca Clínic Barcelona - Institut d’Investigacions Biomèdiques August Pi I Sunyer (IDIBAPS), Barcelona, ESP
5Clínica Universidad de Navarra, Pamplona, Spain
6Department of Hematology, ICAMS, Hospital Clínic de Barcelona, Barcelona, Spain
7Fundació de Recerca Clínic Barcelona-Institut d'Investigacions Biomèdiques August Pi i Sunyer (FRCB-IDIBAPS), Barcelona, Spain
8Fundació de Recerca Clínic Barcelona-Institut d'Investigacions Biomèdiques August Pi i Sunyer (FRCB-IDIBAPS), Barcelona, ESP
9Department of Hematology, Hospital Clínic, Barcelona, Barcelona, Spain
10Fundació de Recerca Clínic Barcelona - Institut d’Investigacions Biomèdiques August Pi i Sunyer (FRCB-IDIBAPS), Bacelona, ESP
11Wellcome-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, United Kingdom
12University of Barcelona, Barcelona, Spain
13Hematopathology Section, Department of Pathology, Hospital Clínic of Barcelona, Barcelona, Spain
14Hematopathology Section, Pathology Department, Hospital Clínic de Barcelona, Barcelona, Spain
15Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), Barcelona, Spain
16Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain
17Universitat de Barcelona, Barcelona, Spain
18Centro de Investigación Biomédica en Red de Cáncer, CIBERONC, Madrid, Spain
19Hospital Clínic de Barcelona, Barcelona, Spain
20Fundació de Recerca Clínic Barcelona - Institut d’Investigacions Biomèdiques August Pi I Sunyer (FRCB-IDIBAPS), Barcelona, Spain

Follicular lymphoma (FL) is the most common indolent B-cell lymphoma. Although achieving a complete response with first-line treatment, most of the patients eventually relapse, which worsens the prognosis and increases the risk of histological transformation to aggressive lymphoma. Therefore, there is the need to better stratify patients at diagnosis and develop tailored therapies for high-risk patients.

Here we assessed the immune profile (Nanostring® nCounter) in formalin-fixed paraffin-embedded (FFPE)-derived biopsies in a series of FL patients at diagnosis, homogenously treated with immunochemotherapy (mostly R-CHOP), and followed at Hospital Clínic de Barcelona. By comparing patients who experienced relapse (n=20) with those who did not relapse after a median follow-up of 14.1 years (n=12), we found 31 differentially expressed genes, 25 of them upregulated in the relapsed group. Among them, we focused on the role of CD70 in FL pathogenesis. CD70 mRNA was higher in patients who eventually relapsed and was associated with inferior progression-free survival (PFS). Moreover, when analyzing the presence of recurrent somatic mutations CD70low cases more frequently bear mutations in CARD11, CIITA, and TNFRSF14 genes.

We next sought to determine the specific subpopulations expressing CD70 by multiplex immunofluorescence (Vectra Polaris). Interestingly, we demonstrated that CD70 is mainly expressed in tumor cells, and it is associated with inferior PFS of the patients. Moreover, a fraction of T cells from the tumor microenvironment also expressed CD70. Intriguingly, higher levels of CD70 were detected in both CD4+ and CD8+ T cells in the relapsed group, including T regulatory and T follicular helper cells. CD70 interacts with CD27, its only known ligand, and can be mutually activated. While CD27 RNA levels were comparable between relapsed and non-relapse groups, when analyzing the CD27 protein, those patients who will eventually relapse, display high CD27 protein levels in B and TFH cells, while low CD27 levels in CD8+ and CD4+ T helper non-follicular cells. Furthermore, we found a correlation between the percentage of CD70+ cells within the B cell population and CD70+ in CD4+ and CD8+ cells (R=0.51, p < 0.05).

To investigate the role of CD70 in FL pathogenesis, we generated CD70 knock-out (CD70-KO) cell lines (n=2) using CRISPR/Cas9. Then T cells from healthy donors were co-cultured with the original FL cell line (CD70+) or the generated CD70-KO FL cells, and we observed that CD70+ tumor cells promoted CD70 expression in T cells. Moreover, CD70 CRISPR/Cas9 in primary samples (n=3), showed that CD70-KO tumor cells exhibit reduced response to proliferative stimuli (CD40L, IL-4 and IL-21). Finally, to advance towards personalized therapies for these high-risk patients and taking advantage of the CAR-T program at our institution, we generated a dual CD19-CD70 CAR-T combining an approved academic product, ARI-0001 (Martinez-Cibrian N, Br J Haematol 2024), and a truncated CD27 protein. We demonstrated that this novel CAR-T is effective in both cell lines and ex vivo primary FL samples, improving tumor depletion in CD19dim samples. Nevertheless, we observed that CAR-T activity is limited by fratricide, which may be overcome by knocking-out CD70 expression in CAR-T cells. Ongoing work includes the optimization of the CRISPR protocol during the CAR-T generation and in vivo studies.

In summary, this multipronged study has allowed us to uncover the prognosis potential of CD70, its biological role in FL patients and the generation of a novel dual CD19-CD70 CAR-T with therapeutic potential in FL as well as other B-NHL.

Disclosures: Mozas: Kyowa Kirin: Honoraria, Other: Travel Grant; AstraZeneca: Honoraria, Other: Travel Grant; Beigene: Honoraria, Other: Travel Grant; Janssen: Honoraria, Other: Travel Grant; Abbvie: Honoraria, Other: Travel Grant; Takeda: Honoraria, Other: Travel Grant. Nadeu: Janssen: Honoraria; AbbVie: Honoraria; AstraZeneca: Honoraria; SOPHiA Genetics: Honoraria; Diagnóstica Longwood: Patents & Royalties: IgCaller algorithm to Diagnóstica Longwood; Gilead: Research Funding. Hodson: Astra Zeneca, GSK: Research Funding. Lopez-Guillermo: Roche: Consultancy, Research Funding; Genmab: Consultancy, Other: Safety Committee clinical trials; Abbvie: Consultancy; Incyte: Consultancy; Janssen: Consultancy; Takeda: Consultancy; Gilead/Kite: Consultancy, Research Funding.

*signifies non-member of ASH