a673 human ewing sarcoma cell line  (ATCC)

Journal: Molecular Therapy Oncology

Article Title: Trabectedin promotes oncolytic virus antitumor efficacy, viral gene expression, and immune effector function in models of bone sarcoma

doi: 10.1016/j.omton.2024.200886

Figure Lengend Snippet: oHSV+trabectedin synergizes to increase the disease control rate and reduce tumor burden in human xenograft models The best response for each treated tumor through 28 days, the average tumor burden, and spider plots tracking individual tumor volumes over the full study period are shown for (A) CHLA-258 (Ewing sarcoma), (B) EW5 (Ewing sarcoma), (C) PDX-0027 (rhabdomyosarcoma), (D) A673 (Ewing sarcoma), and (E) A673 in NSG-SGM3 NK-deficient mice (lack T, B, and NK cells). PBS and oHSV (1.0 × 10 7 pfu) were given intratumorally (i.Tu.) on days 0 and 2. Trabectedin (0.15 mg/kg) was given intravenously (i.v.) on days 0 and 7. Statistical analyses of the disease control rates (CR + PR + SD) were performed using a pairwise Fisher’s exact test with p values adjusted using the Benjamini-Hochberg procedure; ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001. Summarized data with error bars depict mean ± SEM.

Article Snippet: The A673 human Ewing sarcoma cell line (Cat# CCL-81), Vero green monkey kidney cell line (Cat# CRL-1598), and K7M2 mouse osteosarcoma cell line (Cat# CRL-2836) were purchased from the American Type Culture Collection (ATCC) (Manassas, VA).

Techniques: Control

Journal: Molecular Therapy Oncology

Article Title: Trabectedin promotes oncolytic virus antitumor efficacy, viral gene expression, and immune effector function in models of bone sarcoma

doi: 10.1016/j.omton.2024.200886

Figure Lengend Snippet: Trabectedin increases viral intracellular prevalence and decreases antiviral gene expression in immunodeficient Ewing sarcoma models (A) Treatment schema for scRNA-seq tumor collection: PBS and oHSV (1.0 × 10 7 pfu) were given i.Tu. on days 0 and 2, trabectedin (0.15 mg/kg) was given i.v. on day 0, and scRNA-seq samples were collected on day 3 to ensure sufficient cell viability for sequencing ( n = 2 per treatment group). (B) oHSV viral titer (pfu) from A673 tumors 3 days or 6 days following treatment with a single dose of oHSV or oHSV+trabectedin. Error bars indicate standard deviation and the p values from a Student’s unpaired t test between treatment groups at each time point are shown. (C) UMAP plots of oHSV transcript presence in all tumor cells, split by treatment group, from the merged scRNA-seq datasets. (D) Violin plot of the expression level (log-transformed) of all oHSV transcripts for all tumor cells in each treatment group. (E) Expression level of HSV time-dependent gene modules shown as a feature plot (UMAP) for all tumor cells and split by treatment groups that contained oHSV. A relative expression cutoff of 1 minimized color skewing by high-expressing outlier cells and maintained color scale consistency between feature plots. (F) Violin plot of the expression level (log-transformed) of HSV time-dependent gene modules for all tumor cells in treatment groups that contained oHSV. (G) Results from gene set enrichment analysis showing the running enrichment score for the “Herpes simplex virus 1 infection” pathway, which represents the intrinsic cellular response to HSV-1. Gene set enrichment analysis was performed on a gene list which was ranked by the gene expression fold change (log2FC) calculated for tumor cells treated with oHSV+trabectedin compared with those treated with oHSV monotherapy. The schema displays key genes from the “Herpes simplex virus 1 infection” pathway with their respective percent change shown in parentheses (a negative value indicates lower expression in tumors treated with oHSV+trabectedin, as compared to oHSV-treated tumors).

Article Snippet: The A673 human Ewing sarcoma cell line (Cat# CCL-81), Vero green monkey kidney cell line (Cat# CRL-1598), and K7M2 mouse osteosarcoma cell line (Cat# CRL-2836) were purchased from the American Type Culture Collection (ATCC) (Manassas, VA).

Techniques: Gene Expression, Sequencing, Standard Deviation, Expressing, Transformation Assay, Virus, Infection

Journal: Biomedicines

Article Title: 5-Aminolevulinic Acid-Mediated Photodynamic Therapy Potentiates the Effectiveness of Doxorubicin in Ewing Sarcomas

doi: 10.3390/biomedicines10112900

Figure Lengend Snippet: Flow chart of the experimental procedure. Summary of the experimental steps in chronological order, starting from cell seeding to treatment with DOX and/or 5-ALA followed by irradiation. Further measurements were then conducted. Created with BioRender.com. Abbreviations: AFM—atomic force microscope; 5-ALA—5-aminolaevulinic acid; DOX—doxorubicin; ES—Ewing sarcoma; MTS—Cell Proliferation assay; ROS—Cellular Oxidative Stress assay; sec—seconds.

Article Snippet: Three human Ewing sarcoma cell lines, namely: RD-ES (HTB-166™; ATCC, Manassas, VA, USA); A-673 (CRL-1598™; ATCC); and TC-71 (ACC 516; Leibniz Institute DSMZ, Braunschweig, Germany) were employed for all experiments.

Techniques: Irradiation, Microscopy, Proliferation Assay

Journal: Biomedicines

Article Title: 5-Aminolevulinic Acid-Mediated Photodynamic Therapy Potentiates the Effectiveness of Doxorubicin in Ewing Sarcomas

doi: 10.3390/biomedicines10112900

Figure Lengend Snippet: Effect of DOX and 5-ALA PDT on the viability of the ES cell lines and control cells. After 24 h, cells (ES: RD-ES, A-673, TC-71 and MSC) were treated with different concentrations of DOX ( A ) or 5-ALA PDT exposure for 600 sec; ( B ). Negative controls were set to 100% viability and the relative viability was calculated. Mean values were plotted from three independent experiments. Error bars indicate the standard deviations. Statistical significance ( p < 0.05) compared to NT is indicated by ● for RD-ES, * for A-673, ○ for TC-71 and # for MSC. p -values after Bonferroni correction are listed in ( C ) for DOX and ( D ) for 5-ALA PDT. Abbreviations: 5-ALA—5-aminolaevulinic acid; DOX—doxorubicin; ES—Ewing sarcoma; MSC—mesenchymal stem cell; NT—no treatment control; PDT—photodynamic therapy; sec—seconds.

Article Snippet: Three human Ewing sarcoma cell lines, namely: RD-ES (HTB-166™; ATCC, Manassas, VA, USA); A-673 (CRL-1598™; ATCC); and TC-71 (ACC 516; Leibniz Institute DSMZ, Braunschweig, Germany) were employed for all experiments.

Techniques:

Journal: Biomedicines

Article Title: 5-Aminolevulinic Acid-Mediated Photodynamic Therapy Potentiates the Effectiveness of Doxorubicin in Ewing Sarcomas

doi: 10.3390/biomedicines10112900

Figure Lengend Snippet: MTS-viability assessment of ES cell lines after DOX treatment and PDT exposure. For DOX, 15 nM was chosen for all three ES cell lines (RD-ES, A-673, TC-71) as well as MSCs. For the photosensitizer 5-ALA, concentrations adapted to the sensitivity of the cell line were chosen (0.15 mM RD-ES, 0.25 mM TC-71, 0.35 mM A-673, and MSCs). Mock-treated control was set to 100% viability. Error bars indicate the standard deviations (* p < 0.05, ** p < 0.01, *** p < 0.001 relative to other treatment). Abbreviations: 5-ALA—5-aminolaevulinic acid; DOX—doxorubicin; ES—Ewing sarcoma; MSC—mesenchymal stem cell; PDT—photodynamic therapy; sec—seconds.

Article Snippet: Three human Ewing sarcoma cell lines, namely: RD-ES (HTB-166™; ATCC, Manassas, VA, USA); A-673 (CRL-1598™; ATCC); and TC-71 (ACC 516; Leibniz Institute DSMZ, Braunschweig, Germany) were employed for all experiments.

Techniques:

Journal: Biomedicines

Article Title: 5-Aminolevulinic Acid-Mediated Photodynamic Therapy Potentiates the Effectiveness of Doxorubicin in Ewing Sarcomas

doi: 10.3390/biomedicines10112900

Figure Lengend Snippet: ROS assessment of ES cell lines after DOX treatment and PDT exposure. For DOX, 15 nM was chosen for all three ES cell lines (RD-ES, A-673, TC-71) as well as MSCs. For the photosensitizer 5-ALA, concentrations adapted to the sensitivity of the cell line were chosen (0.15 mM RD-ES, 0.25 mM TC-71, 0.35 mM A-673, and MSCs). Results are presented as mean fold difference of mock-treated control. Error bars indicate the standard deviations (* p < 0.05, ** p < 0.01, *** p < 0.001 relative to other treatment, # p < 0.05 compared to untreated control group). Abbreviations: 5-ALA—5-aminolaevulinic acid; DOX—doxorubicin; ES—Ewing sarcoma; MSC—mesenchymal stem cell; PDT—photodynamic therapy; sec—seconds.

Article Snippet: Three human Ewing sarcoma cell lines, namely: RD-ES (HTB-166™; ATCC, Manassas, VA, USA); A-673 (CRL-1598™; ATCC); and TC-71 (ACC 516; Leibniz Institute DSMZ, Braunschweig, Germany) were employed for all experiments.

Techniques:

Journal: Biomedicines

Article Title: 5-Aminolevulinic Acid-Mediated Photodynamic Therapy Potentiates the Effectiveness of Doxorubicin in Ewing Sarcomas

doi: 10.3390/biomedicines10112900

Figure Lengend Snippet: Analysis of Young’s modulus of human ES cell lines and MSC controls. Box plots (medians, minimum, maximum) of the cellular stiffness (Pa) for each cell line is displayed (RD-ES: ( A ) A-673: ( B ) TC-71: ( C ) MSC: ( D ). Outliners are depicted by circles. Statistical significance is indicated by stars over the compared datasets (* p < 0.05, *** p < 0.001). ES cells treated with DOX, PDT, or in combination were stiffer than their corresponding untreated cells, whereas MSCs serving as controls showed no significant change in elasticity. Abbreviations: 5-ALA—5-aminolaevulinic acid; DOX—doxorubicin; ES—Ewing sarcoma; MSC—mesenchymal stem cell; NT—no treatment control; PDT—photodynamic therapy.

Article Snippet: Three human Ewing sarcoma cell lines, namely: RD-ES (HTB-166™; ATCC, Manassas, VA, USA); A-673 (CRL-1598™; ATCC); and TC-71 (ACC 516; Leibniz Institute DSMZ, Braunschweig, Germany) were employed for all experiments.

Techniques: