Fig. 7: Pharmacological reversal of NGFRhi state restores T cell sensitivity.
From: Reversal of pre-existing NGFR-driven tumor and immune therapy resistance
a T cell titration in the presence or absence of tyrphostin (AG-879) in M009R.X1.CL-TR cells. b Quantification of MART-1 T cell cytotoxicity in TR cell lines relative to Ctrl T cells in the presence of absence of AG-879. Error bars represent S.D. of six independent experiments. Statistical analysis by Kruskal–Wallis test; **p < 0.01. c Western blot analysis on D10-TR cells treated with 250 nM of HSP90 inhibitor ganetespib for the indicated days. GAPDH was used as a loading control (see Source data for other replicate). d Quantification of NGFR expression by flow cytometry after 3 days of ganetespib treatment in vitro. Error bars represent S.D. of five independent experiments. Statistical analysis by Mann–Whitney, **p < 0.01. e Colony formation assay of D10-TR cells, treated with control (Ctrl) or MART-1 T cells in a 1:1 ratio and/or 250 nM ganetespib for 72 h. Quantification in f. f Quantification of cytotoxicity in TR cell lines relative to Ctrl T cells (1:1 ratio tumor cell: T cell) in the presence of absence of ganetespib. Two different sets of TCRs are shown. Error bars represent S.D. of six independent replicates. Statistical analysis by Mann–Whitney test; **p < 0.01. g Immunohistochemistry scoring of NGFR on D10-TR tumors after vehicle or 100 mg/kg ganetespib treatment in vivo during 6 days for n = 3 mice. Treatment was given on day 1 and day 4. Statistical analysis by unpaired t-test; *p < 0.05. h Tumor growth curve of D10-TR tumor cells after treatment with Ctrl T cells (n = 5), MART-1 T cells (n = 5), ganetespib 100 mg/kg + Ctrl T cells (n = 7), and ganetespib 100 mg/kg + MART-1 T cells (n = 8). Randomization, T cell injection, and start of ganetespib treatment occurred on day 7. Ganetespib was given twice weekly. Error bars represent S.D. Statistical analysis by Kruskal–Wallis test; ns not significant, *p < 0.05, **p < 0.01. i Survival curve belonging to h. Statistical analysis by Log-Rank Mantel–Cox test, **p < 0.01. Source data are provided as a Source Data file.
