Journal: Cancer Biology & Medicine

Article Title: Nuclear PHGDH regulates macrophage polarization through transcriptional repression of GLUD1 and GLS2 in breast cancer

doi: 10.20892/j.issn.2095-3941.2024.0398

Figure Lengend Snippet: Inhibition of PHGDH enhances the M2-like macrophage polarization phenotype and immunosuppressive function. (A, B) Overexpression and knockdown of PHGDH validated at the mRNA level by qRT-PCR and the protein level by Western blot in THP1 cells. (C, D) Validation of overexpression and knockdown of PHGDH at the mRNA level by qRT-PCR and the protein level by Western blot in BMDM cells. (E, F) Effects of PHGDH overexpression or knockdown in tumor-associated macrophages on polarization function and (G, H) immunosuppressive function ( t -test, * P < 0.05, ** P < 0.01, *** P < 0.001) . (I) Correlation analyses of the transcriptional levels of macrophage-associated markers ( TGF-β and TNF-α ) with PHGDH in human breast cancer, according to data from the TCGA database. (J–L) PHGDH overexpressing BMDMs (OE-PHGDH) and control BMDMs (OE-NC) were mixed with EO771 cells and grafted into C57BL/6 mice ( n = 7). (J) Schematic diagram of the cell transfer experiment and tumor image. (K) Growth curve and tumor weight ( t -test, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns: not significant). (L) Representative immunohistochemistry images of paraffin-embedded xenograft sections. Scale bar, 20 μm.

Article Snippet: The mouse breast cancer cell line EO771 (ATCC, Cat. CRL-3461) was obtained from the ATCC and grown according to the standard protocols on the ATCC website.

Techniques: Inhibition, Over Expression, Knockdown, Quantitative RT-PCR, Western Blot, Biomarker Discovery, Control, Immunohistochemistry

Journal: Journal of Translational Medicine

Article Title: Pharmaceutical targeting Th2-mediated immunity enhances immunotherapy response in breast cancer

doi: 10.1186/s12967-022-03807-8

Figure Lengend Snippet: IPD enhances the antitumor activity of cytotoxic CD8 + T cells in breast cancer. A Representative contour plots and quantification of CD8 + T cells ratios in EO771 and 4T1 tumors (n = 5, t test). B , C Flow cytometry analysis of IFN-γ + and TNF-α + CD8 + T cells from EO771 tumor (B) and 4T1 tumor (C) (n = 5, t test). D Representative immunofluorescence images of CD8 + T cells (green) and granzyme B (red) in EO771 tumors. Scale bar, 25 μm. E , F Representative histogram of Ki-67 expression and percentages of Ki-67 + CD8 + cells in EO771 (E) and 4T1 (F) tumors (n = 5, t test). (G and H) Representative contour plots of PD-1 + Tim-3 + CD8 + T cells of EO771 (G) and 4T1 (H) tumors. Mean ± SEM; ** p < 0.01; *** p < 0.001; ns, not significant

Article Snippet: MDA-MB-231 cell (human breast cancer cell line), EO771 cells (a C57BL/6 murine breast cancer cell line), EMT6 cells (a BALB/c murine breast cancer cell line), and 4T1 cells (a BALB/c murine breast cancer cell line) were obtained from the American Type Culture Collection.

Techniques: Activity Assay, Flow Cytometry, Immunofluorescence, Expressing

Journal: Journal of Translational Medicine

Article Title: Pharmaceutical targeting Th2-mediated immunity enhances immunotherapy response in breast cancer

doi: 10.1186/s12967-022-03807-8

Figure Lengend Snippet: Inhibiting Th2-mediated immunity reshapes the immune landscape in the TME. A , B Representative quantification of myeloid-derived suppressor cell (MDSC), tumor-associated macrophage (TAM), and regulatory T cell (Treg) of CD45 + live cells in EO771 (A) and 4T1 (B) tumors (n = 5, two-way ANOVA). C T-SNE plot of TILs overlaid with color-coded clusters in EO771 tumors. D T-SNE plot of tumor-infiltrating leukocytes overlaid with color-coded clusters from vehicle-treated or IPD-treated mice. E Frequency of clusters of indicated immune cell subsets from vehicle-treated and IPD-treated mice (n = 5, t-test). F Macrophage-associated density t-SNE plots and quantification of an equal number of CD45 + tumor-infiltrating leukocytes in vehicle-treated versus IPD-treated mice. G , H Relative MHC II:CD206 ratio of TAMs in EO771 and 4T1 tumors from vehicle-treated versus IPD-treated mice (n = 5, t test). Mean ± SEM; * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant

Article Snippet: MDA-MB-231 cell (human breast cancer cell line), EO771 cells (a C57BL/6 murine breast cancer cell line), EMT6 cells (a BALB/c murine breast cancer cell line), and 4T1 cells (a BALB/c murine breast cancer cell line) were obtained from the American Type Culture Collection.

Techniques: Derivative Assay

Journal: Journal of Translational Medicine

Article Title: Pharmaceutical targeting Th2-mediated immunity enhances immunotherapy response in breast cancer

doi: 10.1186/s12967-022-03807-8

Figure Lengend Snippet: IPD synergizes with anti-PD1 treatment in breast cancer. A EO771 (left) and 4T1 (right) tumor growth treated with vehicle, IPD, anti-PD1, or IPD + anti-PD1 (n = 5, two-way ANOVA). B Kaplan-Meir survival analysis of 4T1 tumor-bearing mice with indicated treatments (n = 10, log-rank test). C Lung metastatic nodules from 4T1 tumor-bearing mice treated with indicated treatments (n = 5, one-way ANOVA). D Quantification of CD8 + T cells ratios in EO771 and 4T1 tumors treated with indicated treatments (n = 5, one-way ANOVA). E , F Representative flow cytometry images and percentages of IFN-γ + CD8 + T cells from EO771 (E) and 4T1 (F) tumors (n = 5, one-way ANOVA). Mean ± SEM; * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: MDA-MB-231 cell (human breast cancer cell line), EO771 cells (a C57BL/6 murine breast cancer cell line), EMT6 cells (a BALB/c murine breast cancer cell line), and 4T1 cells (a BALB/c murine breast cancer cell line) were obtained from the American Type Culture Collection.

Techniques: Flow Cytometry

Journal: mAbs

Article Title: Activity of murine surrogate antibodies for durvalumab and tremelimumab lacking effector function and the ability to deplete regulatory T cells in mouse models of cancer

doi: 10.1080/19420862.2020.1857100

Figure Lengend Snippet: Complete response rate of anti-mouse PD-L1 clone 80 and anti-mouse CTLA-4 mAbs across eight mouse syngeneic tumor models

Article Snippet: Syngeneic mouse colon cancer (CT26, MC38), breast cancer (EMT6, 4T1, EO771), melanoma (B16F10), renal cell cancer (RENCA), lung cancer (LL/2 lewis lung), or fibrosarcoma (MCA205) tumor cells were engrafted in 6- to 8-week-old female Balb/C (CT26, EMT6, 4T1, RENCA) or female C57BL/6 (B16F10, LL/2 lewis lung, MC38, EO771, MCA205) mice (Envigo).

Techniques:

Journal: mAbs

Article Title: Activity of murine surrogate antibodies for durvalumab and tremelimumab lacking effector function and the ability to deplete regulatory T cells in mouse models of cancer

doi: 10.1080/19420862.2020.1857100

Figure Lengend Snippet: Anti-tumor efficacy and pharmacodynamic profiling of mAbs. Shown are in vivo efficacy and pharmacodynamic effects of anti-mouse PD-L1 and CTLA-4 surrogates in mouse syngeneic tumor models that are sensitive to checkpoint inhibition. (a–c) Kaplan-Meier survival curves of mice engrafted with EMT6, CT26, or MCA205 tumors and treated with anti–PD-L1 or anti–CTLA-4 mAbs. (d–g) Pharmacodynamic activity of monotherapy or combination anti-PD–L1 plus anti–CTLA-4 therapy in EMT6, CT26, and MCA205 models as measured by CD8 or conventional CD4 T-cell proliferation (Ki67 positivity). (h–j) Proportion of FoxP3+ Tregs in tumors, tumor-draining lymph nodes, and spleens of mice engrafted with CT26 tumors

Article Snippet: Syngeneic mouse colon cancer (CT26, MC38), breast cancer (EMT6, 4T1, EO771), melanoma (B16F10), renal cell cancer (RENCA), lung cancer (LL/2 lewis lung), or fibrosarcoma (MCA205) tumor cells were engrafted in 6- to 8-week-old female Balb/C (CT26, EMT6, 4T1, RENCA) or female C57BL/6 (B16F10, LL/2 lewis lung, MC38, EO771, MCA205) mice (Envigo).

Techniques: In Vivo, Inhibition, Activity Assay