![Figure 2. AZD5363 is highly active in vitro and in vivo. (A) AZD5363 treatment induces apoptosis. Five days of inhibitor treatment results in a significant increase of apoptosis in BJAB, whereas no induction of apoptosis is detectable in K422 and HBL-1 cells. Data are expressed as means ± standard deviation of at least 3 independent experiments. (B) AKT inhibitor treatment reduces cell proliferation. CFSE dilutions are measured 3 and 5 days after treatment with AZD5363. In K422 and BJAB cells, proliferation is markedly decreased by AZD5363. Representative results of 3 independent experiments are shown. (C) Tumor growth curves for K422 and WSUDLCL2 xenograft mouse models. AZD5363 treatment (red) significantly reduces tumor growth compared with the vehicle control (blue) in both xenograft mouse models (P = .029 for AZD5363 [n = 9] vs vehicle [n = 8] on day 18 in K422; P = .036 for AZD5363 [n = 9] vs vehicle [n = 9] on day 11 in WSUDLCL2; 1-tailed 2-sample Student t tests). Treatment was initiated after animals developed macroscopic signs of tumors (day 19 after engraftment in K422; day 32 after engraftment in WSUDLCL2). Error bars indicate the standard error of the mean. (D) Western blotting for p-AKT and PTEN expression in the patient-derived GCB DLBCL cell line UPF4D. Lysates from HT and DB cells are used as a positive control for p-AKT and PTEN expression, respectively. (E) AZD8835 and AZD5363 viability assay in UPF4D cells in vitro. PI3Kα/δ inhibition does not affect viability of UPF4D cells compared with the positive control cell line TMD8. In contrast, AKT inhibition induces even stronger cytotoxicity in UPF4D cells compared with the positive control line BJAB. Data are expressed as means ± standard deviation of at least 3 independent experiments. (F) AZD5363 induces apoptosis in UPF4D cells after 24 hours measured by Annexin-V/PI staining. Data are expressed as means ± standard deviation of at least 3 independent experiments. (G) Tumor growth curves of the PDX mouse model WEHA. AZD5363 treatment (red) significantly decreases tumor growth compared with vehicle only treated mice (blue) (P = 7.86 × 10−7 for AZD5363 [n = 9] vs vehicle [n = 9] on day 11; 1-tailed 2-sample Student t test). Treatment was initiated after animals developed macroscopic signs of tumors (day 13 after engraftment). Error bars indicate the standard error of the mean. *P < .05; **P < .01; ***P < .001.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/130/3/10.1182_blood-2016-12-758599/4/blood758599f2.jpeg?Expires=1725167929&Signature=i4uVB13lFBdY7Yys72ZqmUx4yhYo0NyB3miF1h86RTlg9E2wx6NRCOzYm73qYj2vbjshVRWFG6j9XzEdAdpLw4rHFsS-d9iao7JeF1gKCk-7dR2UlgLHIQUFMT94yvjq3jekSmaNIx7w0CUNhSGTtddFCcMDq6CAF4~YA5imtpIqvq76xQV5CYeUxbyPgZX6GyN~JyjhBxE1dlQZKOyql0S4PD5txJtUr4MlCsx7-az~kdmsiyP0UcIqa0CSKncUjLa25jMVIP6JWOy~CcXBZWv0bOMl0l68PdQW1DkFeDfDFCuD-IFXW75rtfS5BdeNFz69-jisRmThN6AHIdRsrQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
AZD5363 is highly active in vitro and in vivo. (A) AZD5363 treatment induces apoptosis. Five days of inhibitor treatment results in a significant increase of apoptosis in BJAB, whereas no induction of apoptosis is detectable in K422 and HBL-1 cells. Data are expressed as means ± standard deviation of at least 3 independent experiments. (B) AKT inhibitor treatment reduces cell proliferation. CFSE dilutions are measured 3 and 5 days after treatment with AZD5363. In K422 and BJAB cells, proliferation is markedly decreased by AZD5363. Representative results of 3 independent experiments are shown. (C) Tumor growth curves for K422 and WSUDLCL2 xenograft mouse models. AZD5363 treatment (red) significantly reduces tumor growth compared with the vehicle control (blue) in both xenograft mouse models (P = .029 for AZD5363 [n = 9] vs vehicle [n = 8] on day 18 in K422; P = .036 for AZD5363 [n = 9] vs vehicle [n = 9] on day 11 in WSUDLCL2; 1-tailed 2-sample Student t tests). Treatment was initiated after animals developed macroscopic signs of tumors (day 19 after engraftment in K422; day 32 after engraftment in WSUDLCL2). Error bars indicate the standard error of the mean. (D) Western blotting for p-AKT and PTEN expression in the patient-derived GCB DLBCL cell line UPF4D. Lysates from HT and DB cells are used as a positive control for p-AKT and PTEN expression, respectively. (E) AZD8835 and AZD5363 viability assay in UPF4D cells in vitro. PI3Kα/δ inhibition does not affect viability of UPF4D cells compared with the positive control cell line TMD8. In contrast, AKT inhibition induces even stronger cytotoxicity in UPF4D cells compared with the positive control line BJAB. Data are expressed as means ± standard deviation of at least 3 independent experiments. (F) AZD5363 induces apoptosis in UPF4D cells after 24 hours measured by Annexin-V/PI staining. Data are expressed as means ± standard deviation of at least 3 independent experiments. (G) Tumor growth curves of the PDX mouse model WEHA. AZD5363 treatment (red) significantly decreases tumor growth compared with vehicle only treated mice (blue) (P = 7.86 × 10−7 for AZD5363 [n = 9] vs vehicle [n = 9] on day 11; 1-tailed 2-sample Student t test). Treatment was initiated after animals developed macroscopic signs of tumors (day 13 after engraftment). Error bars indicate the standard error of the mean. *P < .05; **P < .01; ***P < .001.
