Fig. 1: Characterization of P-BCMA-ALLO1 gene editing. | Nature Communications

Fig. 1: Characterization of P-BCMA-ALLO1 gene editing.

From: TSCM-predominant allogeneic anti-BCMA CAR-T therapy for relapsed/refractory multiple myeloma: preclinical characterization and interim results from a phase 1 trial

Fig. 1

A T cells isolated from HDs were edited using the piggyBac DNA Delivery System and the CC Gene Editing system in a one-step electroporation process. This approach facilitated the stable integration of the CAR into the genome, along with inactivation of TCRαβ and MHC-1. Post-editing, cells were selected, expanded, purified to remove TCR- cells, and cryopreserved for future clinical use. B FCM analysis of CAR-T cells at harvest indicates that the majority of P-BCMA-ALLO1 cells were β2M-TCR-, with further enrichment following TCR- purification. C All P-BCMA-ALLO1 products exhibited high percentages of CAR+, TCR-, and β2M- cells. D The average yield of selected CAR+ cells at harvest was 7.0 ± 4.1 times the initial T cell count. E P-BCMA-ALLO1 cells demonstrated high viability 24 h post-thaw (n = 27 for (C–E). P-BCMA-ALLO1 are significantly less alloreactive compared to donor-matched unedited CAR-T without CC editing (β2M+TCR+) by either (F) the percentage of proliferating CAR-T (CTV-) in co-culture with irradiated PBMC in a MLR assay (n = 3 donors) or (G) the percent killing of CAR-T cells (n = 3 donors/targets) by two lots of donor-specific alloreactive T cells (effectors) at different effector to target (E:T) ratios. Statistical analyses in (F) and (G) was conducted using two-tailed student’s t-test and two-tailed paired t-test, respectively. All error bars in this figure represent standard deviation. FACS contour plots are from a representative sample. *: p < 0.05, **: p < 0.01, ****: p < 0.0001. Data shown are for research lots. Source data are provided as a Source Data file.

Back to article page