Journal: Discover. Oncology

Article Title: Breast cancer stem cells generate immune-suppressive T regulatory cells by secreting TGFβ to evade immune-elimination

doi: 10.1007/s12672-023-00787-z

Figure Lengend Snippet: Tumor-initiating CSCs and immunosuppressive Treg cells have positive correlation in breast cancer. A Plots showing correlation between NANOG and FOXP3, OCT4 and FOXP3, from GSE25066 breast cancer patient dataset (left). Plots demonstrating positive correlation between cancer stemness marker ALDH1A1 and suppressive Treg signature FOXP3 in brain cancer glioma (dataset used: Tumor Brain Lower Grade Glioma (2022-v2)-tcga-532-tpm-gencode36) (middle) and prostate cancer (dataset used: Tumor Prostate Adenocarcinoma-TCGA-497-rsem-tcgars) (right). Datasets were obtained from ‘R2: Genomics Analysis and Visualization Platform’ and ‘TCGA’ database. “r” is correlation co-efficient. B Kaplan-Meier (KM) plot depicting low overall survival (OS) probability of breast tumor patients harboring high expression of CSC marker NANOG and Treg signature gene FOXP3. C Representative Immunohistochemistry (IHC) images showing elevated expression of stemness markers OCT4, SOX2, NANOG, and Treg signature gene FOXP3 in low-grade vs. high-grade breast tumor tissues (n = 4 in each group). Scale bar = 50 µM and magnification 40X. D Bar diagram showing number of OCT4, SOX2, NANOG, and FOXP3 positive cells in low-grade vs. high-grade breast tumor tissues as determined by IHC-staining. E Bar graphs showing incidence of higher cancer stemness markers OCT4, SOX2, NANOG, and ALDH1A1 along with suppressive Treg marker FOXP3 in TNBC tissues than ER + -luminal breast tumor tissues. Bar graphs were plotted using RNA profiling data of TNBC dataset (GSE30682) and ER + -luminal breast tumor dataset (GSE5460) available in ‘R2: Genomics Analysis and Visualization Platform’. F Flow-cytometry data showing percent CD44 + CD24 − CSC population in ER + -luminal MCF-7, triple-negative MDA-MB-231, and MDA-MB-468 breast cancer cells. G Plot showing a positive correlation between ALDH1A1 and FOXP3 in TNBC dataset (GSE76714) available in ‘R2: Genomics Analysis and Visualization Platform’. H Bar graph demonstrating occurrence of higher Treg cell percentage in TNBC patients than non-TNBC patients. Data were represented as the mean ± SD of minimum 3 independent experiments performed in triplicate. Student’s t -test (unpaired) ( D , E , H ) and one-way ANOVA ( F ) was used to assess the data where *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. BC: breast cancer, TNBC: triple-negative breast cancer

Article Snippet: Primary antibody (1:100) against OCT4, SOX2, NANOG, and FOXP3 (Thermofisher Scientific, Cat. No. MA1104, MA1014, MA1017, and PA1806, respectively) were used.

Techniques: Marker, Expressing, Immunohistochemistry, Flow Cytometry

Journal: Discover. Oncology

Article Title: Breast cancer stem cells generate immune-suppressive T regulatory cells by secreting TGFβ to evade immune-elimination

doi: 10.1007/s12672-023-00787-z

Figure Lengend Snippet: CSC-secreted TGFβ induces immunosuppressive Treg cell generation. A Plot showing positive correlation between CSC markers ALDH1A1 and TGFβ1, from GSE69031 breast cancer patient dataset (left) and representative plot demonstrating correlation between TGFβ1 and suppressive Treg marker FOXP3 using GSE5460 breast cancer dataset (right panel). Datasets were obtained from ‘R2: Genomics Analysis and Visualization Platform’ database. “r” is correlation co-efficient. B Plots showing a positive correlation of ALDH1A1 with TGFβ (left panel) and FOXP3 with TGFβ (right panel) in TNBC dataset (GSE142102) available in ‘R2: Genomics Analysis and Visualization Platform’. C Representative bar plots showing level of secreted TGFβ in the CM of MDA-MB-468-derived CSCs and NSCCs, as determined by ELISA. D FACS plots (left panel) and bar diagram (right panel) depicting percent immune-suppressive Treg cells generated from CD4 + T cells following activation by (i) only α-CD3 + α-CD28, (ii) α-CD3 + α-CD28 + TGFβ + IL2, and (iii) α-CD3 + α-CD28 + CSC-CM. E FACS plots (left panel) and bar diagram (right panel) depicting percent Treg cell generation from activated T cells cultured in presence of CSC-CM pre-incubated with or without anti-TGFβ antibody. F Bar plot demonstrating expression of TGFβ per-CSC in control vs. 3-cycles of doxorubicin-treated CSCs derived from MDA-MB-468 spheroids. G Representative bar plot showing the amount of TGFβ present in the CM of control vs. 3-cycles of doxorubicin-treated CSCs derived from MDA-MB-468 spheroids determined by ELISA. Data were represented as the mean ± SD of minimum 3 independent experiments performed in triplicate. Student’s t-test (unpaired) was used to assess the data where *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. CSC: cancer stem cell; MFI: mean fluorescence intensity; CM: conditioned medium

Article Snippet: Primary antibody (1:100) against OCT4, SOX2, NANOG, and FOXP3 (Thermofisher Scientific, Cat. No. MA1104, MA1014, MA1017, and PA1806, respectively) were used.

Techniques: Marker, Derivative Assay, Enzyme-linked Immunosorbent Assay, Generated, Activation Assay, Cell Culture, Incubation, Expressing, Fluorescence