CD19-CART (Shanghai Children's Medical Center)

CD19 CAR-T therapy has achieved remarkable responses in relapsed/refractory non-Hodgkin lymphoma (NHL). However, challenges persist, with refractory responses or relapses after CAR-T administration linked to CD19 loss or downregulation. Given the co-expression of CD19 and BCMA in NHL, we hypothesized that dual targeting could enhance long-term efficacy. We optimized different dual-targeting approaches, including co-transduction of two lentiviral vectors, bicistronic, tandem, and loop and pool strategies, based on our academic anti-CD19 (ARI0001) and anti-BCMA (ARI0002h) CAR-T cells. Comparison with anti-CD19/CD20 or anti-CD19/CD22 dual targeting was also performed. We demonstrate that anti-CD19/BCMA CAR-T cells can be effectively generated through the co-transduction of two lentiviral vectors after optimization to minimize competition for cellular resources. Co-transduced T cells, called ARI0003, effectively targeted NHL tumor cells with high avidity, outperforming anti-CD19 CAR-T cells and other dual-targeting approaches both in vitro and in vivo, particularly in low CD19 antigen density models. ARI0003 maintained effectiveness post-CD19 CAR-T treatment in xenograft models and in spheroids from relapsed CART-treated patients. ARI0003 CAR-T cells were effectively manufactured under Good Manufacturing Practice conditions, with a reduced risk of genotoxicity compared to other dual-targeting approaches. A first-in-human phase 1 clinical trial (CARTD-BG-01; this study was registered at ClinicalTrials.gov [NCT06097455]) has been initiated to evaluate the safety and efficacy of ARI0003 in NHL.

CD19‐targeted chimeric antigen receptor T‐cell (CAR‐T) immunotherapy has transformed the management of relapsed/refractory large B‐cell lymphoma (LBCL), yet durable remissions are observed in less than half of treated patients. The tumor microenvironment (TME) is a key and understudied factor impacting CD19 CAR‐T therapy outcomes. Using NanoString nCounter transcriptome profiling (n = 24) and multiplex immunohistochemistry (mIHC, n = 15), we studied the TME in pretreatment biopsies from patients with LBCL undergoing CD19 CAR‐T therapy. Patients who achieved complete response (CR) after CAR‐T therapy demonstrated higher expression of genes associated with T‐cell trafficking and function, whereas those who did not achieve CR had higher expression of genes associated with macrophages and T‐cell dysfunction. Distinct patterns of immune infiltration and fibrosis in the TME were associated with CAR‐T therapy outcomes, and these findings were corroborated using artificial intelligence‐assisted image analyses. Patients who achieved CR had a lower proportion of the biopsy occupied by an interspersed immune infiltrate and a higher proportion of hypocellular/fibrotic regions. Furthermore, mIHC revealed lower density of CD4+ T cells and higher densities of both macrophages and tumor cells expressing PD‐L1 in non‐CR patients. Spatial analysis revealed that PD‐1+ T cells were in close proximity to PD‐L1+ macrophages or PD‐L1+ tumor cells in patients who did not compared to those who did achieve CR after CAR‐T therapy. These findings suggest that morphologic patterns in the TME and engagement of the PD‐1/PD‐L1 axis in pretreatment biopsies may impact CD19 CAR‐T immunotherapy response in patients with LBCL.