Introduction: CAR T-cell therapies have transformed the treatment (tx) landscape of patients (pts) with LBCL. However, more than 50% of pts experience progression in part due to CD19 tumor antigen escape. Dual targeting of two validated B cell antigens, CD20 and CD19, may overcome relapses associated with antigen loss. JNJ-90014496 (JNJ-4496) is a CD20/CD19 bi-specific CAR T-cell therapy currently under clinical investigation in pts with relapsed/refractory (R/R) LBCL in an ongoing phase 1b study (NCT05421663). Initial results showed objective response (OR) and complete response (CR) rates in CAR T-cell-naive pts of 91% and 75% at all doses, and 96% and 77%, respectively, at the recommended phase 2 dose (RP2D) of 75 million (M) CAR+ T-cells. No Grade 3/4 cytokine release syndrome (CRS) and only 1 Grade 3 immune effector cell-associated neurotoxicity syndrome (ICANS) were observed at the RP2D in 25 pts (Patel K et al., EHA 2025). We report biomarker analyses from pts with R/R LBCL in this phase 1b study and the relationship between these biomarkers and clinical outcomes.

Methods: JNJ-4496 dose levels included a weight-based dose of 2.0M CAR+ T-cells/kg (n=18), a fixed dose of 150M (n=8), and a fixed step-down dose of 75M CAR+ T-cells (n=25). Biomarker analyses conducted in this study include peripheral blood and tumor-based assessments. Peripheral blood assessments included CAR T-cell expansion (quantitative polymerase chain reaction-based transgene and flow cytometry assays); B-cell depletion and recovery (TBNK [BD Biosciences, CA], and custom B-cell flow cytometry panels); and cytokine analysis (V-plex proinflammatory panel [Meso Scale Discovery, MD]). Tumor-based assessments included CD20 and CD19 target expression analysis by immunohistochemistry (IHC). Gene expression analysis using RNA sequencing assessed LymphoMAP tumor microenvironment (TME) archetypes and their association with tx response.

Results: JNJ-4496 cellular kinetics showed comparable CAR T-cellular expansion across all three dose levels tested in 51 pts with R/R LBCL. In addition to the expansion of total CD3+ CAR T-cells, comparable expansion of CD4+ and CD8+ CAR T-cells, and similar B-cell depletion were observed across all doses. Elevated levels of cytokines associated with T-cell cytotoxicity coincided with the onset of CRS or ICANS and correlated with severity grade. Of fixed doses evaluated, overall lower median cytokine levels were observed at the RP2D. Higher median peak CAR T-cell levels were normalized to pre-infusion tumor burden in pts with CR. Tumor samples from 43 pts showed high levels of baseline CD20 expression (H-score >200 in 79% of pts), but variable CD19 expression by IHC (median [interquartile range] 190 [168 to 220]). Using an H-score cutoff of 100, 16% pts had low or heterogenous expression levels of target antigen: 9% had low levels of either CD20 or CD19 expression and 7% showed low levels of both target antigens. OR and CR rates were 100% and 90%, respectively, in 31 pts with both CD20 and CD19 high/medium (H-score >100) and were 82% and 73% in 11 pts with at least 1 antigen low (H-score <100). Initial findings from 24 pts with available gene expression data showed 100% OR rate across all LymphoMAP archetypes, with CR rates in lymph node (92%), fibroblast/macrophage (86%), and T-exhausted (TEX, 80%) archetypes.

Conclusions: Comparable CAR T-cell expansion and peripheral B-cell depletion were observed across all doses. These findings, combined with lower cytokine levels at the 75M fixed dose, which may be indicative of an improved safety profile, support the selection of this dose as the RP2D of JNJ-4496 in pts with R/R LBCL. High response rates were observed in pts with varying levels of CD19 or CD20, including those with low expression levels. In addition, LymphoMAP analysis showed high response rates across archetypes including the TEX archetype, characterized by an abundance of CD8+ T-cells and markers of exhaustion, suggesting that JNJ-4496 may be able to overcome an immunosuppressive TME and provide improved tx response compared with previously approved CD19 CAR T-cell therapies, in which TEX was associated with poor outcomes (Li X, et al. Cancer Cell 2025). As such, these initial findings could potentially explain the high clinical response rates observed with JNJ-4496 in pts with R/R LBCL and will be further evaluated in additional pts recruited in this and future studies.

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2025
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