Article Figures & Data
Figures
- Figure 1.
BRAF inhibition is associated with increased melanoma antigen expression in tumors of patients with metastatic melanoma. Tumors were harvested and mRNA levels of gp100, MART-1, TYRP-1, and TYRP-2 in patients with metastatic melanoma undergoing treatment with a selective inhibitor of BRAFV600E were assayed. A, expression levels of the respective MDAs are shown as fold increase over pretreatment value and are plotted on a log scale in a box and whiskers plot (n = 12). The bottom and top of the 25th and 75th percentile, respectively, for all patients, with the bar indicating the median value. The whiskers indicate the extremes with open circles representing data points greater than 1.5 times the interquartile range. Immunohistochemistry (B; ×40 magnification) and immunofluorescence (C) staining for the melanoma antigen MART-1 in pretreatment and on-treatment biopsies (patient 2) was conducted to confirm that protein expression correlated with mRNA expression. All microscopy was repeated at least 3 times with representative examples shown. P values are from a 2-tailed Student t test with a μ of 1, which corresponds to no change in mRNA levels on treatment. *, P ≤ 0.05.
- Figure 2.
BRAF inhibition is associated with increased CD8+ T-cell infiltrate in tumors of patients with metastatic melanoma. Tumors biopsied pretreatment and on-treatment were stained for H&E and immunohistochemistry (IHC) was conducted for CD8+ T cells with a representative patient shown (A). CD8+ T-cell counts were conducted in a blinded fashion by a dedicated dermatopathologist (B). Average CD8+ T-cell counts are plotted for each patient with error bars representing the standard deviation of four measurements. CD8+ counts from all patients (n = 11) are expressed in a box and whiskers plot both pre- and on-treatment (C).
- Figure 3.
BRAF inhibition is associated with decreased immunosuppressive cytokines and markers of T-cell cytotoxicity but increased T-cell exhaustion markers and PDL1 in tumors of patients with metastatic melanoma. Tumors were harvested and mRNA levels of IL-6, IL-8, IL-10, and TGF-β (A), perforin (n = 11), GranzymeB (n = 11), TIM-3 (n = 14) and PD1 (n = 14; B), and PDL1 (n = 11; C) in patients with metastatic melanoma undergoing treatment with a selective inhibitor of BRAFV600E were assayed. All patients are expressed in a box and whiskers plot. Open circles represent data points greater than 1.5 times the interquartile range. P values indicated are from a 2-tailed Student t test with a μ of 1, which represents no change in mRNA value with respect to the pretreatment value. *, P ≤ 0.05. Immunohistochemistry (×40 magnification) for the perforin, GranzymeB, and TIM-3 (patient 6; D) and PDL1 (patient 12; E) in pretreatment and on-treatment biopsies was conducted to confirm that protein expression correlated with mRNA expression. The dotted line = tumor–stroma interface and the inset is the isotype-specific control.
- Figure 4.
Melanoma antigen expression and CD8+ T-cell infiltrate are decreased at time of progression and restored through MEK inhibition. A, tumors were harvested at time of progression and at time of treatment with combined BRAF inhibition and MEK inhibition for patient 3. mRNA levels of the melanoma antigens gp100, MART-1, TYRP-1, and TYRP-2 were assayed. B, immunohistochemistry (IHC) was conducted for CD8+ T cells on patient tumor samples. C, CD8+ T cells were identified and counted by a dedicated pathologist. Average CD8+ T-cell counts are plotted with error bars representing the SD of 4 measurements. *, P ≤ 0.05.
Additional Files
Supplementary Data
Files in this Data Supplement:
- Supplementary Figure Legend - PDF file - 69K
- Supplementary Figure 1 - PDF file - 112K, Inhibition of BRAFV600E increases expression of MART-1 in tumors of patients with metastatic melanoma.
- Supplementary Figure 2 - PDF file - 349K, Inhibition of BRAFV600E is not associated with a significant increase in HLA expression in patients with metastatic melanoma.
- Supplementary Figure 3 - PDF file - 19K, Melanoma antigen expression is decreased at time of progression.
- Supplementary Table 1 - PDF file - 50K, Patient Characteristics.
- Supplementary Table 2 - PDF file - 30K, Inhibition of BRAFV600E increases expression of MDAs in tumors of patients with metastatic melanoma.
Volume 19, Issue 5
