Extended Data Fig. 10: Characterization of the PDX PA1443 and RP-6306/gemcitabine in combination drive tumor regression in CCNE1-amplified CDX models.
From: CCNE1 amplification is synthetic lethal with PKMYT1 kinase inhibition
a, Distribution of the gene-level copy number in the CCNE1-amplified pancreatic cancer (PA1443) patient-derived xenograft (PDX). Highlighted is the amplicon containing CCNE1. b, Whole cell lysates from FT282-hTERT TP53R175H parental (WT) and CCNE1-high (C3) cell lines and PA1443 PDX tumor tissue were immunoblotted with antibodies to cyclin E and vinculin. Representative of two immunoblots. c, Tumor tissues of PA1443 PDX implanted in BALB/c nude mice were prepared for FFPE and stained with a cyclin E1 antibody. d, Changes in body weight in tumor-bearing PA1443 PDX implanted in BALB/c nude mice treated either with RP-6306 or vehicle. RP-6306 was administered orally BID at 2.5 mg kg−1 for the duration of the experiment. Results are expressed as mean % body weight ± s.e.m. (n = 8). P value relative to vehicle was determined with a one-way ANOVA test. Only P values < 0.05 are indicated. ns, not significant (P value > 0.05). e, g, Same data as presented in Fig. 5d, e with tumour growth values plotted in log scale for OVCAR3 (e) and HCC1569 (g). The percent regression of the RP-6306 10 mg kg−1 and Gemcitabine 20 mg kg−1 combination arm is indicated. Results are expressed as mean tumor volume ± s.e.m. (OVCAR3 n = 7 (vehicle), 6 (10 mg kg−1 RP-6306), 7 (20 mg kg−1 gemcitabine), 7 (10 mg kg−1 RP-6306 + 20 mg kg−1 gemcitabine); HCC1569 n = 7 (vehicle), 7 (10 mg kg−1 RP-6306), 7 (20 mg kg−1 gemcitabine), 7 (10 mg kg−1 RP-6306 + 20 mg kg−1 gemcitabine)). Please refer to Fig. 5 for TGI and P values. f, h, Growth traces for individual OVCAR3 (f) and HCC1569 (h) tumors from the experiments shown in Fig. 5d, e from mice treated with the gemcitabine/RP-6306 combination. For comparison, tumor growth data for two mice treated with vehicle are shown. i, j, Changes in body weight in tumor-bearing OVCAR3 (i) and HCC1569 (j) CB-17 SCID and SCID-beige mice treated with either RP-6306, gemcitabine or both. Gemcitabine was delivered once weekly intraperitoneally and RP-6306 was given orally BID for 21 d after which all treatments were stopped, and body weight monitored for the remainder of the experiment. Results are expressed as mean ± s.e.m. (OVCAR3 n = 7 (vehicle), 6 (10 mg kg−1 RP-6306), 7 (20 mg kg−1 gemcitabine), 7 (10 mg kg−1 RP-6306 + 20 mg kg−1 gemcitabine); HCC1569 n = 7 (vehicle), 7 (10 mg kg−1 RP-6306), 7 (20 mg kg−1 gemcitabine), 7 (10 mg kg−1 RP-6306 + 20 mg kg−1 gemcitabine)) . Also indicated are P values relative to vehicle were determined with a one-way ANOVA test. Only P values < 0.05 are indicated. For gel source data, see Supplementary Fig. 1.
