rorγ inverse agonist sr2211  (Tocris)

Journal: bioRxiv

Article Title: RORγ bridges cancer-driven lipid dysmetabolism and myeloid immunosuppression

doi: 10.1101/2023.11.19.567414

Figure Lengend Snippet: a, Experimental design: adult wt C3H/HeJ mice were fed with HCD for 8 weeks before being injected with K1735-M2 tumor cells. When the tumor volume reached ∼0.80-1.00 cm 3 , typically 20-24 days after tumor cell injection, primary tumors were surgically removed. Twenty-eight days after tumor removal, mice were sacrificed for sample collection and analysis. SR2211 was administered to a first group of mice 10 days before tumor removal (SR2211_PRE). A second group of mice received SR2211 treatment 5 days after tumor removal (SR2211_POST). b, Lung metastatic areas (n=5) in HCD-fed K1735-M2 mice treated with either SR2211 or vehicle control. Representative images are shown. Scale bar, 1mm. c, Lung metastatic areas in NCD- or HCD-fed MN/MCA1 wt mice treated with vehicle, SR2211, anti-PD1, or SR2211 plus anti-PD1 (n=5). Representative images are shown. Scale bar, 1mm. One experiment was performed. Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. One-way ANOVA with Tukey’s multiple comparisons test.

Article Snippet: Starting from day 10 after tumor cell injection, MN/MCA1-bearing mice received intraperitoneal treatment with the following reagents: anti-IL6 monoclonal antibody (BioXcell, Clone MP5-20F3) at a dose of 200 μg twice a week; anti-IL1 (anakinra, Sobi, Stockholm, Sweden) at a dose of 200 μg twice a week; anti-PCSK9 monoclonal antibody mAb1 (kindly provided by Amgen Inc, Thousand Oaks, CA, USA) at a dose of 200 μg per mouse twice a week (MN/MCA1 tumor) or once a week (KP mice); anti-CSF1R monoclonal antibody (BioXcell, Clone AFS98) at an initial dose of 400 μg per mouse, followed by twice-weekly doses of 200 μg for the duration of the experiment; anti-CCR2 inhibitor (Tocris) at a dose of 75 μg per mouse twice a week; and the RORγ inverse agonist SR2211 (Tocris) at a dose of 50 μg per mouse twice a week.

Techniques: Injection, Control

Journal: bioRxiv

Article Title: RORγ bridges cancer-driven lipid dysmetabolism and myeloid immunosuppression

doi: 10.1101/2023.11.19.567414

Figure Lengend Snippet: MN/MCA1 mice were fed NCD or HCD. a-d, Frequency of RORγ + CCR2 + cells and relative RORγ expression levels in: a, M-MDSCs from tumors (n=6); b, lung IMs (n=4), AMs (n=4), and M- MDSCs (n=5); c, BM M-MDSCs (n=5); d, Spleen M-MDSCs (n=5). e, Frequency of RORγ + CCR2 - cells and relative RORγ expression levels in lung AMs (n=4), BM or spleen M-MDSCs (n=5). f, Frequency of RORγ + CMPs and GMPs cells and relative RORγ expression levels (n=4). g, HEK293 cells were co-transfected with CMV-RORγ and IL17A_prom-Luc plasmids. Plasmid 1: RORγ gene is constitutively expressed under the control of CMV promoter. Plasmid 2: Luciferase reporter gene is under the control of the minimal promoter of Il17a gene, whose transcription is driven by RORγ. h-j, HEK293 cells, co-transfected with plasmids 1 and 2, were stimulated with SR0987, desmosterol, cholesterol, or LDL ( h ), or with sera from NCD- or HCD-fed MN/MCA1 TB or TF mice (n=4) ( i ), or exposed to single or combinations of cholesterol, SR2211 and TCM ( j ). Luciferase activity was quantified as relative light units (RLU). Data are representative of five ( a, b, d ), two ( c, e, h-j ), or three ( f ) independent experiments. Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 between selected relevant comparisons. a-f, Unpaired two-tailed t -test; h-j, One-way ANOVA with Tukey’s multiple comparisons test.

Article Snippet: Starting from day 10 after tumor cell injection, MN/MCA1-bearing mice received intraperitoneal treatment with the following reagents: anti-IL6 monoclonal antibody (BioXcell, Clone MP5-20F3) at a dose of 200 μg twice a week; anti-IL1 (anakinra, Sobi, Stockholm, Sweden) at a dose of 200 μg twice a week; anti-PCSK9 monoclonal antibody mAb1 (kindly provided by Amgen Inc, Thousand Oaks, CA, USA) at a dose of 200 μg per mouse twice a week (MN/MCA1 tumor) or once a week (KP mice); anti-CSF1R monoclonal antibody (BioXcell, Clone AFS98) at an initial dose of 400 μg per mouse, followed by twice-weekly doses of 200 μg for the duration of the experiment; anti-CCR2 inhibitor (Tocris) at a dose of 75 μg per mouse twice a week; and the RORγ inverse agonist SR2211 (Tocris) at a dose of 50 μg per mouse twice a week.

Techniques: Expressing, Transfection, Plasmid Preparation, Control, Luciferase, Activity Assay, Two Tailed Test

Journal: bioRxiv

Article Title: RORγ bridges cancer-driven lipid dysmetabolism and myeloid immunosuppression

doi: 10.1101/2023.11.19.567414

Figure Lengend Snippet: a-h , NCD- or HCD-fed MN/MCA1 wt mice treated with the RORγ inhibitor SR2211 or vehicle control (n=4). a, Lung metastatic area; representative images are shown, scale bar, 1 mm; b, tumor growth (volume and weight); c, total blood cholesterol; d, FACS quantification of TAMs and relative CCR2 expression; e , TNFα, MHC-II, and CD206 expression by TAMs (MFI); f, frequency of IM (top) and AM (bottom) subsets and relative expression (MFI) of CCR2, MHC-II, and CD206; g, CMP and GMP frequencies in BM; h , IFNγ, PD-1 and CTLA4 expression (MFI) in intratumoral CD8 + T cells. i-l, HCD-fed K1735-M2-bearing wt mice treated with SR2211 or vehicle control. i, Tumor weight (n=6); j, FACS quantification of TAMs and tumor-infiltrating M-MDSCs (n=4); k, IM and AM frequencies and relative TNFα expression (MFI) (n=5); l, IFNγ, PD1, and CTLA4 expression (MFI) by lung CD8 + T cells (n=5). m, Tumor growth (volume and weight) in NCD- or HCD-fed wt mice treated with vehicle, SR2211, anti-PD1, or SR2211 plus anti-PD1 (n=5). One experiment was performed. Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 between selected relevant comparisons. a, b (right) , c-h, k, l, m (right), One-way ANOVA with Tukey’s multiple comparisons test. i, j, Unpaired two-tailed t -test. b (left) , m (left), Two-way ANOVA with Tukey’s multiple comparisons test

Article Snippet: Starting from day 10 after tumor cell injection, MN/MCA1-bearing mice received intraperitoneal treatment with the following reagents: anti-IL6 monoclonal antibody (BioXcell, Clone MP5-20F3) at a dose of 200 μg twice a week; anti-IL1 (anakinra, Sobi, Stockholm, Sweden) at a dose of 200 μg twice a week; anti-PCSK9 monoclonal antibody mAb1 (kindly provided by Amgen Inc, Thousand Oaks, CA, USA) at a dose of 200 μg per mouse twice a week (MN/MCA1 tumor) or once a week (KP mice); anti-CSF1R monoclonal antibody (BioXcell, Clone AFS98) at an initial dose of 400 μg per mouse, followed by twice-weekly doses of 200 μg for the duration of the experiment; anti-CCR2 inhibitor (Tocris) at a dose of 75 μg per mouse twice a week; and the RORγ inverse agonist SR2211 (Tocris) at a dose of 50 μg per mouse twice a week.

Techniques: Control, Expressing, Two Tailed Test

Journal: Cell

Article Title: A multiscale map of the stem cell state in pancreatic adenocarcinoma

doi: 10.1016/j.cell.2019.03.010

Figure Lengend Snippet: (A and B) 3D growth of KPf/fC cells (A) and KPR172H/+C cells (B) in the presence of the SR2211 or vehicle (n=3).

Article Snippet: In vivo and in vitro drug therapy The RORγ inverse agonists SR2211 (Cayman Chemicals, 11972, or Tocris, 4869) was resuspended in DMSO at 20 mg/ml or 50 mg/ml, respectively, then mixed 1:20 in 8% Tween80-PBS prior to use.

Techniques:

Journal: Cell

Article Title: A multiscale map of the stem cell state in pancreatic adenocarcinoma

doi: 10.1016/j.cell.2019.03.010

Figure Lengend Snippet: (A-B) Analysis of flank KPf/fC tumor-bearing NSG mice treated with SR2211 or vehicle for 2 weeks. Strategy (A). Flank tumor growth following treatment with vehicle or SR2211 for 2 weeks (B). Fold change in tumor volume relative to volume at enrollment (n=4–6).

Article Snippet: In vivo and in vitro drug therapy The RORγ inverse agonists SR2211 (Cayman Chemicals, 11972, or Tocris, 4869) was resuspended in DMSO at 20 mg/ml or 50 mg/ml, respectively, then mixed 1:20 in 8% Tween80-PBS prior to use.

Techniques: