Journal: Frontiers in Oncology

Article Title: HER3 Differentiates Basal From Claudin Type Triple Negative Breast Cancer and Contributes to Drug and Microenvironmental Induced Resistance

doi: 10.3389/fonc.2020.554704

Figure Lengend Snippet: Differential HER3 response to AKT and PI3K inhibitors in Basal and Claudin type TNBC. (A) Biochemical assessment of the effect of AKT (GDC0068) and PI3K (GDC0077) inhibition in a panel of Basal and Claudin type Triple Negative Breast Cancer (TNBC) showing expression and upregulation of total and phosphorylated HER3 and downstream signaling in the Basal type TNBC cells (BT-20, HCC70, and MDA-MB-468). Cells were treated for 48h with the respective kinase inhibitors at 1uM (B) HER3 quantification in the TNBC cell lines tested in A and their corresponding mutational status of PTEN, PI3K, KRAS, and BRAF (C) . (D) Biochemical signalling analysis in response to NRG- 1α and -1β in MDA-MB-468 showing increased signalling with NRG- 1β isoform.

Article Snippet: Cells were seeded into 96-well plates followed by treatment with individual kinase inhibitors GDC0068 (Selleckchem), GDC0077, Gefitinib (Selleckchem), Neratinib (Selleckchem), Lapatinib (LC Laboratories), Sapitinib (FisherScientific), and the HER3 Ab (R&D Systems), in the presence or absence of NRG-1α or -1β.

Techniques: Inhibition, Expressing

Journal: Frontiers in Oncology

Article Title: HER3 Differentiates Basal From Claudin Type Triple Negative Breast Cancer and Contributes to Drug and Microenvironmental Induced Resistance

doi: 10.3389/fonc.2020.554704

Figure Lengend Snippet: Neuregulin but not Hepatocyte Growth Factor decreases sensitivity to kinase inhibitors in Basal type Triple Negative Breast Cancer (TNBC). (A) Viability assay assessing the effect of Neuregulin-1β (50 ng/ml) in the presence of the small molecule inhibitors GDC0068 (1uM) and GDC0077 (1uM) after 96h treatment in a panel of TNBC cell lines in BT-20, HCC-70, MDA-MB-468, BT-549, and MDA-MB-231 and (B) Dose dependent effects of NRG- 1α and -1β isoforms [Low Dose (LD) 5 ng/ml, Medium Dose (MD) 50 ng/ml, and high dose (HD) 100 ng/ml] in the presence or absence of GDC0068 (1uM) or GDC0077 (1uM) for 96h in HCC-70, MDA-MB-468, and MDA-MB-231 TNBC. (C) Effect of HGF treatment (50 ng/ml) after 96h in the presence or absence of GDC0068/GDC0077 at 1uM. Drug response graphs and Heat map graphs show CellTiter-Glo luminescence viability measurements at the end of the experiments compared to untreated control. Experiments were performed in triplicate. Data are means ± SD]. Drug response graphs were analyzed using two-way analysis of variance (ANOVA)/Tukey’s multiple comparison test, *<0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001].

Article Snippet: Cells were seeded into 96-well plates followed by treatment with individual kinase inhibitors GDC0068 (Selleckchem), GDC0077, Gefitinib (Selleckchem), Neratinib (Selleckchem), Lapatinib (LC Laboratories), Sapitinib (FisherScientific), and the HER3 Ab (R&D Systems), in the presence or absence of NRG-1α or -1β.

Techniques: Viability Assay, Control, Comparison

Journal: Frontiers in Oncology

Article Title: HER3 Differentiates Basal From Claudin Type Triple Negative Breast Cancer and Contributes to Drug and Microenvironmental Induced Resistance

doi: 10.3389/fonc.2020.554704

Figure Lengend Snippet: Gefitinib in combination with AKT/PI3K inhibitors sensitizes Basal type Triple Negative Breast Cancer (TNBC). (A) HCC-70, MDA-MB-468, and MDA-MB-231 TNBC cells treated in the presence of NRG-1β (50 ng/ml) does not significantly decrease sensitivity to gefitinib (200 nM) after 96 h. (B) Viability assay of TNBC cells as in (A) treated with the small molecule inhibitors GDC0068 (1uM), GDC0077 (1uM), gefitinib (200 nM), or a combination of the drugs +/- NRG1β (50 ng/ml) for 96h. In the HCC-70 and MDA-MB-468 cell lines, combination therapy demonstrated decreased viability compared to the MDA-MB-231 cell line which showed no significant difference in response, regardless of treatment (C) Biochemical analysis of the p/EGFR and p/HER3 showing decreased but not abolished phosphorylated proteins indicating a potent and sustained signaling in presence of NRG1β in the Basal type TNBC cells, HCC-70, and MDA-MB-231; Cells were treated for 48h at the drug concentrations shown in (B) . The Claudin type MDA-MB-231 cell line showed no HER3 phosphorylation. Drug response graphs show CellTiter-Glo luminescence viability measurements at the end of the experiments compared to untreated control and analyzed using two-way analysis of variance (ANOVA)/Tukey’s multiple comparison test, *<0.05, ***P<0.001]. Experiments were performed in triplicate. Data are means ± SD].

Article Snippet: Cells were seeded into 96-well plates followed by treatment with individual kinase inhibitors GDC0068 (Selleckchem), GDC0077, Gefitinib (Selleckchem), Neratinib (Selleckchem), Lapatinib (LC Laboratories), Sapitinib (FisherScientific), and the HER3 Ab (R&D Systems), in the presence or absence of NRG-1α or -1β.

Techniques: Viability Assay, Phospho-proteomics, Control, Comparison

Journal: Frontiers in Oncology

Article Title: HER3 Differentiates Basal From Claudin Type Triple Negative Breast Cancer and Contributes to Drug and Microenvironmental Induced Resistance

doi: 10.3389/fonc.2020.554704

Figure Lengend Snippet: Pan HER targeting sensitizes Basal type but not Claudin type Triple Negative Breast Cancer (TNBC) cells to AKT and PI3K inhibition. (A) Biochemical assessment of downstream signaling in the PI3K/AKT signaling pathway after combination therapy with neratinib (300 nM) and GDC0068 (1 uM) or GDC0077 (1 uM) after 48 h, showing deceased signaling in the Basal TNBC cell lines but not in the Claudin type TNBC. (B) Viability assay of TNBC cells treated with the small molecule inhibitors GDC0068, GDC0077, neratinib, or a combination of the drugs for 96h [drug treatments as in (A) ]. In the BT-20, HCC-70, and MDA-MB-468 cell lines, combination therapy demonstrated decreased viability compared to the MDA-MB-231and BT-549 cell line which showed no added benefit of combination therapy. Drug response graphs show CellTiter-Glo luminescence viability measurements at the end of the experiments compared to untreated control and analyzed using two-way analysis of variance (ANOVA)/Tukey’s multiple comparison test, *<0.05, **P < 0.01, ***P<0.001]. Experiments were performed in triplicate. Data are means ± SD].

Article Snippet: Cells were seeded into 96-well plates followed by treatment with individual kinase inhibitors GDC0068 (Selleckchem), GDC0077, Gefitinib (Selleckchem), Neratinib (Selleckchem), Lapatinib (LC Laboratories), Sapitinib (FisherScientific), and the HER3 Ab (R&D Systems), in the presence or absence of NRG-1α or -1β.

Techniques: Inhibition, Viability Assay, Control, Comparison

Journal: Neoplasia (New York, N.Y.)

Article Title: PIK3CA mutant cervical cancer is selectively suppressed by PI3Kα inhibition (Alpelisib/BYL-719 and Inavolisib/GDC-0077) and cooperates with HPV directed T cell therapy

doi: 10.1016/j.neo.2026.101305

Figure Lengend Snippet: Inhibition of cervical cancer cell lines by PI3K α inhibitors. The maximum dose for each cell line was selected, and the percentage inhibition was compared with the corresponding control. (A) Alpelisib (BYL-719). (B) Inavolisib (GDC-0077). All cell lines have PIK3CA mutations except SiHa. **** = p < 0.0001 by one-way ANOVA and Dunnett’s multiple comparison test. <20% inhibition interpreted as no or negative response.

Article Snippet: GDC-0077 was also purchased from MedChem Express (Cat# HY-101562.

Techniques: Inhibition, Control, Comparison