Journal: Cell Reports Medicine
Article Title: Multimodal targeting of metastatic renal cell carcinoma via CD70-directed allogeneic CAR-NKT cells
doi: 10.1016/j.xcrm.2025.102321
Figure Lengend Snippet: Biomarker profiling of primary RCC patient samples reveals therapeutic potential for CAR-NKT cell treatment (A) Experiment design to profile primary RCC patient tumor samples using flow cytometry. 15 RCC primary samples were included for analyses. (B) Bar graphs showing the cell cluster proportions of the 15 primary RCC tumor samples. (C) Comparison of mRNA expression levels in RCC tumor cells versus normal kidney tissues (upper bars) and corresponding log2 fold change between tumor and normal tissues (lower bars). Data were obtained from The Human Protein Atlas (HPA). nTPM, normalized transcript per million. (D) CD70 mRNA expression in various human cancer types. Raw data were collected from the Genomic Data Commons (GDC). FPKM-UQ, fragments per kilobase of transcript per million mapped reads-upper quartile. (E–I) Biomarker profiling of RCC tumor cells. (E) Fluorescence-activated cell sorting (FACS) detection of CD70 expression on tumor cells. Data from three representative samples are shown. CD70 hi indicates high (≥50%) CD70 expression; CD70 med/low indicates medium to low (<50%) CD70 expression. (F) Quantification of CD70 expression among the 15 primary RCC tumor cell samples. (G) FACS detection of NK ligand expression on tumor cells. Data from one representative sample are shown. (H) Quantification of NK ligand expression among the 15 primary RCC tumor cell samples. (I) Comparison of NK ligand expression between CD70 hi ( n = 9) and CD70 med/low ( n = 6) tumor cell samples. (J–O) Biomarker profiling of RCC TME cells. (J) FACS detection of PD-1 expression and Granzyme B and Perforin production in T cells. TAM/MDSC low , samples with <20% TAM/MDSCs among total immune cells; TAM/MDSC high , samples with >20% TAM/MDSCs among total immune cells; Iso, isotype staining. (K) Comparison of PD-1 expression and Granzyme B and Perforin production in T cells between TAM/MDSC low ( n = 6) and TAM/MDSC high ( n = 9) RCC samples. (L) FACS detection of CD1d expression on the indicated RCC TME cells. (M) Quantification of (L) ( n = 15). (N) FACS detection of CD70 expression on the indicated RCC TME cells. (O) Quantification of (N) ( n = 15). (P) Schematics showing the multimodal targeting of RCC tumor, microenvironment, and alloreactive T cells by CAR70-NKT cells. Data are presented as the mean ± SEM. ns, not significant; ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗∗ p < 0.0001 by Student’s t test (I and K) or one-way ANOVA (M and O).
Article Snippet: Gene mRNA expression levels in RCC tumor cells versus normal kidney tissues , The Human Protein Atlas , https://www.proteinatlas.org/about/download.
Techniques: Biomarker Discovery, Flow Cytometry, Comparison, Expressing, Fluorescence, FACS, Staining