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1594 NOTCH1 Mutations Are Associated with Therapy-Resistance in Patients with B-Cell Lymphoma Treated with CD20xCD3 Bispecific Antibodies

Program: Oral and Poster Abstracts
Session: 621. Lymphomas: Translational – Molecular and Genetic: Poster I
Hematology Disease Topics & Pathways:
Research, Adult, Bispecific Antibody Therapy, Translational Research, Non-Hodgkin lymphoma, Lymphomas, Bioinformatics, B Cell lymphoma, Diseases, Treatment Considerations, Biological therapies, Lymphoid Malignancies, Technology and Procedures, Study Population, Human
Saturday, December 7, 2024, 5:30 PM-7:30 PM

Emil Ramsø Kyvsgaard1,2,3*, Morten Grauslund4*, Lene Sjö5,6*, Linea Melchior7*, Trine Grantzau4*, Lise Mette Gjerdrum8*, Trine Trab9*, Lærke Sloth Andersen10*, Anne Ortved Gang, MD11*, Marie Fredslund Breinholt12*, Michael Boe Møller13*, Jacob Haaber Christensen, MD, PhD14*, Thomas Stauffer Larsen, MD, PhD15,16*, Michael Roost Clausen, MD, PhD17,18*, Anna Caroline Hasselbalch Riley, MD1,19*, Carsten Utoft Niemann, MD, PhD19,20*, Kirsten Gronbaek, MD, PhD, PR20,21,22,23, Martin Hutchings, MD, PhD19,24 and Simon Husby, MD, PhD2*

1Phase 1 Unit, Rigshospitalet, Copenhagen, Denmark
2Rigshospitalet, Department of Hematology, Copenhagen, Denmark
3Grønbæk Group, Biotech Research and Innovation Centre, BRIC, University of Copenhagen, Copenhagen, Denmark
4Dept, of Pathology, Rigshospitalet, Copenhagen, Denmark, Copenhagen, DNK
5Department of Pathology, Rigshospitalet, Copenhagen, Denmark
6University of Copenhagen, Copenhagen, DNK
7Dept. of Pathology, Rigshospitalet, Copenhagen Ø, DNK
8Zealand University Hospital, Roskilde, DNK
9Dept. Of Hematology, Rigshospitalet, Copenhagen, Denmark, Copenhagen, DNK
10Dept. of Hematology, Rigshospitalet, Copenhagen, Denmark, Copenhagen, DNK
11Copenhagen University Hospital, Herlev, Denmark
12Dept. of Pathology, Herlev Hospital, Copenhagen, Denmark
13Pathology, Odense University Hospital, Odense, Denmark
14Dept. of Hematology, Odense University Hospital, Odense, Denmark
15Department of Hematology, Odense University Hospital, Odense, Denmark, Odense, Denmark
16Department of Clinical Research, University of Southern Denmark, Odense, Denmark
17Aarhus University Hospital, Vejle, DNK
18Vejle Hospital, Vejle, Denmark
19Department of Hematology, Rigshospitalet, Copenhagen, Denmark
20Department of Clinical Medicine, Faculty of Health Science, University of Copenhagen, Copenhagen, Denmark
21Rigshospitalet, University of Copenhagen, Copenhagen N, Denmark
22Department of Hematology, Copenhagen University Hospital, Rigshospitalet, Copenhagen, Denmark
23The Danish Stem Cell Center (Danstem), Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
24Department of Clinical Medicine, University of Copenhagen, Copenhagen, Denmark

Introduction:

CD20xCD3 bispecific antibody therapy (BsAb) have been approved for treatment of relapsed/refractory diffuse large B cell lymphomas (DLBCL) and follicular lymphoma (FL) with ≥2 prior lines of treatment. Approximately 50% of patients do not achieve long lasting remission when treated with single agent CD20xCD3 BsAb. Mechanisms of treatment resistance to these novel T-cell engaging antibodies are unclear. Previously, tumor mutations in TP53 have been associated with poor response to both immunochemotherapy (i.e. R-CHOP) and CD19 CAR-T cell therapy in patients with B-cell lymphoma. Genetic tumor aberrations have not yet been examined thoroughly in patients with long follow-up treated with CD20xCD3 BsAb.

Methods:

We conducted a retrospective study including patients with R/R B cell lymphoma treated at Rigshospitalet Copenhagen and Vejle University Hospital, both in Denmark from 2017-2023 as part of phase 1/2 clinical trials with CD20/CD3-targeted BsAb. Pre-treatment formalin-fixed and paraffin-embedded (FFPE) archival specimens were collected from 106 patients, of which 56 had sufficient tumor involvement and DNA quantity for clinical grade diagnostic next-generation sequencing (NGS). For analysis of clonal development under CD20xCD3 BsAb exposure, we collected paired relapse biopsies from 17 patients. A custom NGS lymphoma gene-panel covering 59 genes recurrently mutated in lymphoid malignancies were used with a median coverage >10,000x for all samples.

Results:

We examined pre-treatment tumors from 56 patients with B-cell NHL who received CD20xCD3 BsAbs from 2017-2023. Median age was 70 years, 60.7% were male, and the median number of prior lines of therapy was three. Median follow-up time was 24.2 months.

The most frequently altered genes in pre-treatment tumors were CREBBP (47%), KMT2D (47%), TP53 (30%), TNFRSF14 (23%), BCL2 (25%), and GNA13 (18%).

In a univariate Cox model examining mutations in pre-CD20xCD3 BsAb treatment biopsies, NOTCH1 mutations were associated with significantly inferior progression-free survival (PFS) (crude hazard ratio (HR): 3.46, p = 0.026), while TP53 mutations did not impact overall response rate (ORR) or survival.

Examining CRS grade ≥2, we found that mutations in GNA13 (n =10) and CARD11 (n=9) were negatively associated with CRS grade ≥2 (p = 0.004 and p = 0.006). When adjusting for multiple testing, both mutations remained significant (q = 0.060 and q = 0.067). Mutations in CARD11 or GNA13 were not associated with inferior overall response rate (ORR) or PFS.

The most common mutations at relapse after CD20xCD3 exposure were CREBBP (41%), STAT6 (24%), BCL2 (24%), NOTCH1 (24%), KMT2D (18%) and BCL2 (18%).

Analyzing clonal evolution during CD20xCD3 BsAb, we identified gross expansion of NOTCH1 mutated clones in 29% of the paired relapse biopsies (these NOTCH1 mutations were undetectable in pre-treatment biopsies). Of the four new NOTCH1 clones identified at relapse, three were in exon 34. All exon 34 clones were detected in germinal center B cell like DLBCL samples. All patients with NOTCH1-mutated tumors (either in biopsy before or after CD20xCD3 BsAb; total 8 patients [14%] of the cohort) died within 6 months of lymphoma progression after initial detection of NOTCH1 aberration. Surprisingly, three patients were observed to have diminishing TP53 clone size at post-CD20xCD3 BsAb relapse.

Conclusions: NOTCH1 mutations predict poor survival and are a resistance mechanism to CD20xCD3 BsAb, while TP53 mutations do not seem to have an adverse effect on response or survival. We suggest investigation of NOTCH1 mutations in patients before undergoing treatment with CD20 targeted T-cell engaging therapies.

Disclosures: Grauslund: Amgen, Astra Zeneca, Thermo Fisher Scientific: Consultancy; Merck: Research Funding. Gjerdrum: consultancy fees from Kyowa Kirin: Consultancy. Trab: Janssen: Research Funding. Larsen: Roche: Consultancy; Gilead: Consultancy; Genentech: Research Funding. Clausen: AbbVie: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Travel Expenses; Gilead: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; AstraZeneca: Consultancy, Honoraria, Other: Travel Expenses; Genmab: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Travel Expenses; Incyte: Consultancy, Honoraria; Roche: Consultancy, Honoraria, Other: Travel Expenses; Pfizer: Other: Travel Expenses; Janssen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Travel Expenses. Niemann: Novo Nordisk: Research Funding; CSL Behring, Genmab, Takeda, Beigene, MSD, Lilly: Consultancy; AbbVie, Janssen, AstraZeneca, Novo Nordisk Foundation, Octapharma: Consultancy, Research Funding. Gronbaek: Otsuka Pharma: Membership on an entity's Board of Directors or advisory committees; Janssen: Research Funding. Hutchings: AstraZeneca: Membership on an entity's Board of Directors or advisory committees, Research Funding; Takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; BMS/Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; AbbVie: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Genmab: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Genentech: Research Funding; Incyte: Research Funding; Janssen/J&J: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Merck: Honoraria, Membership on an entity's Board of Directors or advisory committees; Novartis: Research Funding.

*signifies non-member of ASH