CD20-specific nanoCAR T-cells incubated with CD70+ JD033T organoid line

CD70-specific nanoCAR T-cells incorporating 4-1BB as costimulatory domain show the highest efficacy

CD70-specific nanoCAR T-cells incubated with CD70+ JD033T organoid line

Patients with newly diagnosed large B-cell lymphoma (LBCL) and International Prognostic Index (IPI) scores ≥3 have inferior outcomes with rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP). We evaluated efficacy and safety of epcoritamab plus R-CHOP in newly diagnosed LBCL in Arm 1 of the phase 1b/2 EPCORE NHL-2 trial. Adults with CD20+ LBCL and IPI scores 3-5 received R-CHOP (6 cycles) plus epcoritamab for a total of 1 year of treatment. Primary endpoint was investigator-assessed overall response rate (ORR). Key secondary endpoints included complete response (CR) rate, duration of CR (DOCR), progression-free survival (PFS), overall survival (OS), minimal residual disease (MRD) negativity, and safety. Forty-seven patients were treated. Median age was 64 years, 89% had Ann Arbor stage IV, and 53% had bulky disease (≥7 cm). ORR was 98% (95% CI, 89-100) and CR rate 85% (95% CI, 72-94). At median follow-up of 44.2 months, median DOCR, PFS and OS were not reached (NR; 95% CI, NR-NR); estimated 2-year PFS and OS rates were 80% (95% CI, 65-89) and 87% (95% CI, 73-94), respectively. MRD-negativity was achieved in 100% of MRD-evaluable patients. Most common adverse events were neutropenia (74%), anemia (72%), and cytokine release syndrome (CRS; 60%). CRS events were predominantly grade 1-2; all resolved. Epcoritamab plus R-CHOP induced high response rates in patients with newly diagnosed LBCL and IPI scores 3-5. These promising findings support the ongoing phase 3 evaluation of this treatment regimen. This trial was registered at ClinicalTrials.gov (NCT04663347).

There is an unmet need for effective, off-the-shelf therapies for relapsed or refractory aggressive B cell non-Hodgkin lymphoma (B-NHL). Part 2 of the current study was an open-label, nonrandomized, phase 1 study of escalating doses of the CD19-4-1BBL co-stimulatory molecule, englumafusp alfa, in combination with glofitamab in patients with relapsed or refractory B-NHL. Obinutuzumab pretreatment was administered 7 days before the first glofitamab dose. Glofitamab step-up dosing in cycle 1 was followed by 11 cycles of glofitamab plus englumafusp alfa. Englumafusp alfa was administered at escalating doses, with the initial dose on cycle 2 day 8 (C2D8) or cycle 1 day 10 (C1D10). Primary objectives were to establish the maximum tolerated dose, and safety and tolerability. A total of 134 patients were enrolled, including 109 with aggressive B-NHL and 25 with indolent B-NHL. The maximum tolerated dose of englumafusp alfa was not reached; one dose-limiting toxicity occurred (grade 5 Pneumocystis jirovecii pneumonia). Adverse events were reported in 98.5% of all patients, with grade 3/4 adverse events in 59.0%. Grade 5 adverse events occurred in ten patients. In the subgroup of C2D8 patients with aggressive B-NHL (n = 83), overall response and complete metabolic response rates were 68.7% and 56.6%, respectively; among those without previous exposure to chimeric antigen receptor T cell therapy (n = 41), the corresponding rates were 73.2% and 65.9%. Pharmacodynamic changes following englumafusp alfa administration supported its co-stimulatory mode of action. These data demonstrate that the addition of englumafusp alfa to glofitamab is associated with encouraging efficacy and robust pharmacodynamic modulation, as well as a safety profile consistent with glofitamab monotherapy, in patients with relapsed or refractory B-NHL. CTIS identifier: 2022-502616-37-00 ; ClinicalTrials.gov identifier: NCT04077723 .

Differentially expressed genes in nanoCAR T-cells from cluster CD8WT vs CD4WT

BACKGROUND:Circulating nucleic acids in blood plasma form an attractive, minimally invasive resource to study human health and disease. In this study, we aimed to identify cell-free RNA alterations that can distinguish cancer patients from cancer-free individuals. METHODS:We first performed mRNA capture sequencing on 266 blood plasma samples from cancer patients and controls, including a discovery set of 208 donors across 25 cancer types and a replication set of 58 donors across three cancer types. We first conducted group-level comparisons and then compared individual patient profiles to a reference control population in a one-versus-many approach. This approach was further evaluated in independent cohorts: a prostate cancer plasma cohort (n = 180), a non-malignant disease plasma cohort (n = 125), a lymphoma plasma cohort (n = 65), and a bladder cancer urine cohort (n = 24), each including both patients and controls. RESULTS:Here we show that cancer patients exhibit both cancer type-specific and general cell-free RNA alterations. However, differentially abundant RNAs vary widely among patients and across cohorts, hampering robust biomarker identification. By comparing individual patient profiles to control populations, we identify so-called biomarker tail genes, which strongly deviate from controls. The number of these genes per sample distinguishes cancer patients from control samples. Independent cohorts also confirm the potential of this approach. CONCLUSIONS:Our findings demonstrate substantial heterogeneity in cell-free RNA alterations among cancer patients and propose that patient-specific changes can be exploited for classification.

CD70KO improves the functionality of CD70-specific nanoCAR T-cells.

Abstract The purpose of this study was to assess whether transcripts exhibiting strongly deviating abundance in plasma mRNA profiles can reliably differentiate prostate cancer from non-cancer states. To do so, we focused on biomarker tail genes (BTG), defined as protein-coding genes whose cell-free transcript abundance in an individual sample deviates by at least three standard deviations from a healthy control reference distribution. We applied the BTG identification and classification workflow to blood plasma samples from individuals with newly diagnosed prostate cancer (n = 132; 62 early-stage, 70 late-stage) and healthy donors (n = 48). Classification thresholds were established using a train-test cross validation approach (70%), and performance was evaluated in held-out validation samples (30%) and in a separate non-malignant cohort including healthy donors (n=37) and patients with non-malignant conditions (n=88). Across training and validation analyses, a consensus set of 247 prostate cancer BTG enabled discrimination between prostate cancer samples and healthy controls, with sensitivity and specificity reaching 100% in the validation cohort of male donors. Of note, the number of BTG per plasma sample was not associated with disease stage, and classification remained highly accurate in age-matched subsets, indicating limited influence of age on results. When applied to a cohort of individuals with diverse non-malignant conditions, the established BTG threshold yielded 94.4% specificity, and none of patients with benign prostatic hyperplasia were misclassified (n=5). To explore redundancy within the BTG set, we evaluated both cluster-derived subsets and algorithmically selected minimal panels. Multiple small subsets, including a ten-gene panel identified by a greedy selection strategy, achieved perfect classification within the validation cohort, demonstrating that strong discriminatory power is retained even when BTG sets are substantially reduced. In conclusion, blood plasma BTG constitute a highly accurate signature for prostate cancer detection, and the ability of small BTG subsets to reproduce full-set performance highlights opportunities for targeted assays. Further evaluation in broader populations and across cancer stages will refine the potential of BTG-based approaches for early detection and monitoring. (The last two authors contributed equally to this work) Citation Format: Annelien Morlion, Philippe Decruyenaere, Kathleen Schoofs, Jasper Anckaert, Nickolas J. Ramirez, Justine Nuytens, Eveline Vanden Eynde, Kimberly Verniers, Celine Everaert, Guy Brusselle, Steven Callens, Filomeen Haerynck, Dimitri Hemelsoet, Eric Hoste, Jo Lambert, Nicolaas Lumen, Fritz Offner, Koen Paemeleire, Vanessa Smith, Lies Van den Eynde, Jo Van Dorpe, Amber Vanhaecke, Hans Van Vlierberghe, An Mariman, Olivier Thas, Jo Vandesompele, Pieter Mestdagh. Biomarker tail genes in blood plasma cell-free RNA enable accurate detection of prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 2601.

BACKGROUND:Chemotherapy-free regimens, such as rituximab and lenalidomide, are attractive first-line treatment options for follicular lymphoma, but combinations providing deeper, more durable responses are needed. We aimed to assess 3-year activity and safety of epcoritamab, a subcutaneously administered CD3 × CD20 bispecific antibody, plus rituximab-lenalidomide as first-line treatment for follicular lymphoma. METHODS:EPCORE NHL-2 is an open-label, phase 1b/2, clinical trial. Arm 6 of the study was conducted at 20 hospitals in six countries in Europe (Spain, the Netherlands, Czech Republic, Sweden, Belgium, and Denmark) and the USA in patients aged 18 years and older with CD20+, histologically confirmed grade 1-3A follicular lymphoma, and an Eastern Cooperative Oncology Group performance status of 0-2. Patients received intravenous rituximab 375 mg/m2 once per week in cycle 1 (28 days per cycle) and every 4 weeks in cycles 2-6; oral lenalidomide 20 mg daily on days 1-21 of cycles 1-12; and subcutaneous epcoritamab in a two-step-up dosing regimen of a 0·16 mg priming dose, followed by a 0·8 mg intermediate dose, then full 48 mg doses in cycle 1, 48 mg once per week in cycle 2, and 48 mg every 4 weeks in subsequent cycles for up to 2 years. The primary endpoint was investigator-assessed overall response rate by Lugano Response Criteria for Malignant Lymphoma. The full analysis set and the safety set included all patients who received one or more doses of trial drug. This study is registered with ClinicalTrials.gov (NCT04663347) and is ongoing (closed to new participants). FINDINGS:Between Sept 23, 2021, and May 3, 2022, 45 patients were assessed for eligibility in Arm 6, 41 of whom were eligible and included in the full analysis and safety sets. 21 (51%) of 41 patients were male and 20 (49%) were female; 27 (66%) of patients were White, one (2%) was Asian, one (2%) was of another race, and 12 (29%) did not have race reported. At a median follow-up of 35·9 months (IQR 35·8-36·6) the overall response rate was 95% (95% CI 84-99; 39 of 41 patients). The most common grade 3-4 adverse events were neutropenia (20 [49%] of 41 patients), infections (12 [29%]), COVID-19 (six [15%]), and alanine aminotransferase increase (five [12%]). Serious adverse events occurred in 29 (71%) patients, including cytokine release syndrome (13 [32%]), serious infections (13 [32%]; one grade 2), pyrexia (three [7%]), organising pneumonia (one [2%]), pleural effusion (one [2%]), pneumonitis (one [2%]), pulmonary embolism (one [2%]), maculopapular rash (two [5%]), toxic skin eruption (one [2%]), atrial fibrillation (one [2%]), diarrhoea (one [2%]), dehydration (one [2%]), ovarian epithelial cancer (one [2%]), cerebrovascular accident (one [2%]), and thrombophlebitis (one [2%]). Treatment-related deaths occurred in three (7%) patients due to septic shock, progressive multifocal leukoencephalopathy, and COVID-19. INTERPRETATION:Epcoritamab plus rituximab-lenalidomide showed high activity in first-line treatment for follicular lymphoma, with 95% of patients having a response over 3 years of follow-up, supporting this treatment regimen as a promising approach that warrants further analysis to ascertain its role in this setting. FUNDING:Genmab and AbbVie.

Differentially expressed genes in bulk RNAseq data from CD70-4_1BB:ζ KO vs CD70-4_1BB:ζ WT

CD70KO has no effect on CD20-specific nanoCAR T-cells.

Differentially expressed genes per cluster

CD70 WT and KO CD70-specific nanoCAR T-cells have a distinct transcriptional profile after production.

Differentially expressed genes in nanoCAR T-cells from cluster CD8KO vs CD4KO

The treatment of relapsed/refractory (R/R) diffuse large B-cell lymphoma (DLBCL) remains challenging, with inadequate responses to salvage chemoimmunotherapy limiting patients’ ability to receive potentially curative treatments like autologous stem cell transplantation (ASCT). Epcoritamab, a subcutaneous CD3×CD20 bispecific antibody, has demonstrated antitumor activity in R/R DLBCL as a monotherapy and in combination with chemotherapy. In Arm 4 of the EPCORE® NHL-2 phase 1b/2 trial (NCT04663347), transplant-eligible patients with CD20+ R/R DLBCL received epcoritamab plus rituximab, dexamethasone, cytarabine, oxaliplatin/carboplatin (R-DHAX/C). Patients could continue epcoritamab until ASCT or progression. Twenty-nine patients received epcoritamab plus R-DHAX/C; 72% had stage IV disease; 66% had primary refractory disease. As of January 15, 2025 (median follow-up 40.4 months), overall response rate (primary endpoint) was 79%, and complete response rate was 69%. Sixteen patients (55%) proceeded to ASCT and five remained on epcoritamab monotherapy. At 36 months, an estimated 70% of responses were ongoing, 59% of patients were progression-free, and 76% were alive. Common treatment-emergent adverse events (TEAE) were thrombocytopenia (90%), anemia (66%), and neutropenia (59%). Cytokine release syndrome occurred in 45% of patients; all were grade 1–2 and resolved after a median of 2 days. Immune effector cell-associated neurotoxicity syndrome occurred in one patient. No fatal TEAE or clinical tumor lysis syndrome were observed. Epcoritamab plus R-DHAX/C achieved deep, durable responses with manageable safety. Over half of patients proceeded to ASCT, a potentially curative treatment. These findings suggest the potential of epcoritamab combined with standard chemoimmunotherapy as an effective salvage treatment for patients with R/R DLBCL.

CD70 expression on nanoCAR T-cells.

Differentially expressed genes in nanoCAR T-cells from cluster CD4KO vs CD4WT

Differentially expressed genes in nanoCAR T-cells from cluster CD8KO vs CD8WT

Background Tisagenlecleucel is approved in the United States and Europe for adults with r/r FL after ≥2 lines of prior therapy. We report the final analysis from the phase 2 ELARA trial (NCT03568461) with >5-y median follow-up (mFU). Methods Pts with r/r FL (grade 1-3A) and ≥2 prior lines of systemic therapy (including an anti-CD20 monoclonal antibody [Ab] and alkylating agent) received a single tisagenlecleucel infusion (0.6-6 × 108 CAR+ viable T cells). Bridging therapy was permitted. Long-term efficacy and safety outcomes were evaluated. Cellular kinetics were assessed by qPCR. Results As of May 28, 2025, 97 pts were infused (mFU, 61.0 mo [range: 3.1–67.0]). At baseline, pts with high-risk (HR) disease included 60% with high FLIPI score of ≥3, 62% with POD24, 64% with bulky disease (>7 cm or 3 lesions >3 cm), 68% with double refractory to prior CD20 Ab and alkylating agent, and 21% with high tumor burden. The ORR (86.2%) and CRR (68.1%) were consistent with prior data (Dreyling M, Blood 2024). The mDOR was not reached (NR; 95% CI: 35.8–NE); estimated 4-y DOR in all responders was 61% (95% CI, 48.8–71.1) and 71.2% (95% CI, 57.7–81.1) in pts with CR. The mPFS was 53.2 mo (95% CI,18.2–NE); estimated 5-y PFS was 46% (95% CI, 35.0–56.3) in all infused pts and 59.8% (95% CI, 46.2–71.1) in pts with CR. In pts with HR disease, 5-y estimated PFS was 35.5% (high FLIPI), 41.1% (POD24), 45.1% (bulky disease), 50.5% (double refractory). In pts with high tumor burden, 5-y PFS was NE due to small subgroup size. Among all infused pts, median OS and estimated 5-y OS were NR and 74.1% (95% CI, 63.0–82.3). Among pts with HR disease, estimated 5-y OS was 64.4% (high FLIPI), 74.6% (POD24), 71.1% (bulky disease), 79.8% (double refractory), and 65.5% (high tumor burden). CAR transgene persistence (Tlast) was observed up to 60.9 mo; median Tlast was 8.6 mo (range: 0.6–60.9). Blood and lymphatic system disorders occurring >1 y after infusion were reported in 11 (13.1%) pts (neutropenia [6.0%], anemia [4.8%], and thrombocytopenia [3.6%]), and infection and infestations in 34 (40.5%) pts (COVID-19 [17.9%] and pneumonia [11.9%]). A total of 11 second primary malignancies were reported in 7 (7.2%) pts: 2 events each of basal cell carcinoma, myelodysplastic syndrome, squamous cell carcinoma of skin, and 1 event each of acute myeloid leukemia, bladder transitional cell carcinoma, Bowen’s disease, malignant melanoma, and metastatic squamous cell carcinoma. In total, 22 pts died during the study due to disease progression (n=8), AEs (n=13, mostly infections), and euthanasia. Conclusions After >5 y mFU, tisagenlecleucel continues to demonstrate durable responses and prolonged survival in pts with r/r FL including pts with HR disease. No new safety signals were reported. More than 75% of pts were alive, and approximately half remained progression-free at this final analysis, indicating the curative potential of tisagenlecleucel in r/r FL.