TEST:

nCounter® CAR-T Characterization Panel

"In vivo efficacy was evaluated using a systemic NALM-6 challenge in immune-deficient NSG mice at HIP CD19 CAR T cell doses of 5e5 and 5e6 cells per animal...Outperforming donors also showed reduced expression of MHC class II of alpha and beta heterodimers (HLA-DQA1 and HLA-DQB1). The information collected across phenotypic and functional comparisons will be compared with clinical performance."

Conclusions Our data suggest that the poor outcome of some LBCL pts receiving CAR T is due to the presence of high monocytes levels expressing a four-gene signature in an inflammatory microenvironment. Additionally, the pre-manufacturing combined analysis of the gene signature and of PB AMC, could be used to plan trials evaluating CAR T cells versus other newly approved therapies such as bispecific antibodies.

Discussion Our study shows that anti-BCMA CAR-T therapy has provided apparent therapeutic advantage over the existing drugs in response rate and long-term survival. Compared with medullary disease, BCMA-specific CAR T cell-based therapy showed limited and discrepant efficacy in EMD, perhaps due to the the properties of EMD-specific microenvironment.

Interestingly, expression of top genes in CAR T from one the non-response group (A07) was different than other group although it showed good response in ex vivo cytotoxicity.While the three individual assays by themselves would not be sufficient to predict for response, taken together, the multiple criteria from ex vivo cytotoxicity assay, T cell phenotypic analysis and gene expression analysis is capable of identifying response to this second generation CAR T product. This study will continue to analyze the CAR T from upcoming phase II clinical trial and perform robust corelative analysis to predict the clinical outcomes based on multiple characteristics of CAR T products while striving to identify statistical weight behind these assays to develop a more reliable prediction model through use of machine learning.

Our study highlights that unmanipulated leukapheresis share peculiar immunophenotypic and transcriptional features that correlate with CAR T-cell expansion and survival of pts treated with Tisa-cel and Axi-cel. Despite these results warrant functional validation and confirmation in larger cohort (ongoing), the gene signature identified may represent a pre-manufacturing predictive biomarker of CAR T-cell efficacy that might preferentially drive their employment versus other newly approved therapies, such as bispecific T-cell engagers, at the single patient level.

We anticipate that this information will prove useful across many aspects of the development, manufacturing and clinical application of cellular therapies, and we hope that the findings will lead to improvements in the safety and efficacy of cell therapy products. Data from this project will be made publicly available to the scientific community.