Figure 2 from GPR65 Inactivation in Tumor Cells Drives Antigen-Independent CAR T-cell Resistance via Macrophage Remodeling
Year: 2025
Type: other
Abstract: Cross-species integrative analysis ranks GPR65 as the top CAR T response driver in B-ALL. A, Venn diagram of the CaRS derived independently by comparing h.CR (responders, n = 19) and h.NR (nonresponders, n = 8) from bone marrow RNA-seq profiles of patients and RNA-seq of in vitro cultured mouse m.PR and m.CR tumor cells (n = 3 per group). Statistical significance of overlap computed using the two-tailed Fisher exact test. B, Principal component analysis plot integrating human and mouse RNA-seq profiles performed using the expression of 307 human CaRS genes. PC, principal component. C, S-curve of combined gene ranking using a total of 8,116 genes for human (h.CR vs. h.NR) and mouse (m.CR vs. m.PR; responders vs. nonresponders). The top four upregulated and downregulated genes are highlighted, and the top hit GPR65 is highlighted in red. D, Gene expression heatmap of 41 genes shared between human and mouse CaRS in both patient and mouse responders (h.CR and m.CR) and nonresponders (h.NR and m.PR) ranked by combined Z-score from C. GPR65 is highlighted in red. E, ROC curve plot for evaluation of the shared CaRS (41 genes) score to predict the response of patients with B-ALL and B-ALL mouse to CD19 CAR T-cell therapy. CI, confidence interval. F, GSEA plot of GPR65 targets from bone marrow RNA-seq profiles of B-ALL patient responders vs. nonresponders treated with CD19 CAR T cells. Responders (N = 19) are defined as patients who had durable (>1 year) complete remissions, and nonresponders (N = 8) are defined as patients who did not respond to treatment and had no evidence of CD19 antigen loss. P value was estimated using the Wilcoxon rank-sum test. G, Boxplot of GPR65 activity calculated using the NetBID2 algorithm using RNA-seq profiles of adult patients with B-ALL treated with blinatumomab. Responders (N = 19) are patients who achieved complete remission by morphology using standard International Working Group criteria, and nonresponders (N = 20) are patients who did not respond to blinatumomab therapy and had no evidence of CD19 antigen loss. P value was estimated using the Wilcoxon rank-sum test. Boxplot shows a summary of the data interims of the minimum, maximum, sample median, and the first and third quartiles. H, qRT-PCR analysis showing relative Gpr65 levels in m.PR and m.CR tumor cells. Representative of three experiments. Statistical significance was calculated using an unpaired t test. All error bars represent mean ± SEM. ****, P < 0.0001.
Language: English
Topic: CAR-T cell therapy research
Subfield: Oncology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W4410047571