Journal: Veterinary Sciences

Article Title: Tumor Growth Progression in Ectopic and Orthotopic Xenografts from Inflammatory Breast Cancer Cell Lines

doi: 10.3390/vetsci8090194

Figure Lengend Snippet: Tumor growth parameters of IPC-366 and SUM149 cell lines in ectopic and orthotopic models.

Article Snippet: The human triple-negative inflammatory breast cancer cell line SUM149 was obtained from Asterand, Inc. (Detroit, MI, USA), (RRID: CVCL_3422).

Techniques: Injection

Journal: Veterinary Sciences

Article Title: Tumor Growth Progression in Ectopic and Orthotopic Xenografts from Inflammatory Breast Cancer Cell Lines

doi: 10.3390/vetsci8090194

Figure Lengend Snippet: IPC-366 and SUM149 xenotransplanted mice, paraffin sections, H-E. ( A ) IPC-366 ectopic xenotransplanted mice. Neoplastic cells arranged in solid masses separated by a scant fibrovascular stroma infiltrating the adjacent dermis (inset: neoplastic cells infiltrating adjacent dermis). ( B ) IPC-366 orthotopic mice. Unencapsulated and densely cellular mass extending into the adjacent adipose tissue. ( C , D ) Ectopic and orthotopic IPC-366 xenotransplanted mice. Tumors are composed of highly pleomorphic cells with marked anisocytosis and anisokaryosis. Binucleated cells are commonly seen (arrow). ( E , F ) Ectopic and orthotopic SUM149 xenografted mice. Solid tumors infiltrate the dermis and adipose tissue. No histological differences were found between the types of SUM149 xenografts. ( G ) Orthotopic SUM149 xenograft. Medium to large round cells with a moderate eosinophilic cytoplasm and large nuclei with one or more evident nucleoli. ( H ) Orthotopic SUM149 xenograft. Presence of neoplastic cells with an elongated and empty cytoplasm that displaced the nuclei to the periphery, suggestive of endothelial-like cells (ELCs) (arrow). Atypical mitoses were frequently seen (arrowhead).

Article Snippet: The human triple-negative inflammatory breast cancer cell line SUM149 was obtained from Asterand, Inc. (Detroit, MI, USA), (RRID: CVCL_3422).

Techniques:

Journal: Veterinary Sciences

Article Title: Tumor Growth Progression in Ectopic and Orthotopic Xenografts from Inflammatory Breast Cancer Cell Lines

doi: 10.3390/vetsci8090194

Figure Lengend Snippet: Estrogen receptor (ER), Progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER-2) expression on ectopic and orthotopic xenografts from IPC-366 and SUM149 cell lines.

Article Snippet: The human triple-negative inflammatory breast cancer cell line SUM149 was obtained from Asterand, Inc. (Detroit, MI, USA), (RRID: CVCL_3422).

Techniques: Expressing

Journal: Veterinary Sciences

Article Title: Tumor Growth Progression in Ectopic and Orthotopic Xenografts from Inflammatory Breast Cancer Cell Lines

doi: 10.3390/vetsci8090194

Figure Lengend Snippet: Steroid hormone secretion studied (P4, DHEA, A4, T, DHT, E1SO4, and E2), on ectopic (subcutaneous) and orthotopic (mammary fat pad) models of IPC-366 and SUM149 xenografts.

Article Snippet: The human triple-negative inflammatory breast cancer cell line SUM149 was obtained from Asterand, Inc. (Detroit, MI, USA), (RRID: CVCL_3422).

Techniques:

Journal: Molecular Cancer Therapeutics

Article Title: Liposome-Encapsulated Eribulin Shows Enhanced Antitumor Activity over Eribulin for Combination Therapy with Anti–PD-1 Antibody

doi: 10.1158/1535-7163.MCT-22-0475

Figure Lengend Snippet: Antitumor activity of eribulin and eribulin-LF in immunodeficient versus immunocompetent mice. A, Antitumor activity of eribulin and eribulin-LF in 4T1#31 subcutaneous transplantation model in immunodeficient and immunocompetent mice. When the average TV reached 80 to 100 mm 3 , tumor-bearing mice were intravenously injected with eribulin (Eri) or eribulin-LF (Eri-LF) at 0.3 mg/kg on a Q7D×2 ( n = 6 per group). Data are presented as mean + SEM. **, P < 0.01 and ****, P < 0.0001 by two-way repeated-measures ANOVA with Tukey multiple-comparisons test using log-transformed values, and statistical results of the difference of TV on the indicated days are shown. B, Comparison of the antitumor activity of eribulin and eribulin-LF in immunodeficient and immunocompetent mice. The ΔT/C value (% of control for Δgrowth) on the last day of measurement was calculated ( n = 6 per group). *, P < 0.05; **, P < 0.01; ***, P < 0.001; and ****, P < 0.0001 by one-way ANOVA with Tukey multiple-comparisons test.

Article Snippet: 4T1 mouse breast cancer cells (ATCC, catalog no. CRL-2539, RRID:CVCL_0125) were purchased from the ATCC and cultured in RPMI1640 medium supplemented with 10% FBS and penicillin–streptomycin.

Techniques: Activity Assay, Transplantation Assay, Injection, Transformation Assay, Comparison, Control

Journal: Molecular Cancer Therapeutics

Article Title: Liposome-Encapsulated Eribulin Shows Enhanced Antitumor Activity over Eribulin for Combination Therapy with Anti–PD-1 Antibody

doi: 10.1158/1535-7163.MCT-22-0475

Figure Lengend Snippet: Antitumor activity of eribulin-LF + anti–PD-1 Ab. A, Antitumor activity of eribulin-LF + anti–PD-1 Ab or each monotherapy in the 4T1#31 subcutaneous transplantation model. When the average TV reached 90 to 100 mm 3 , mice bearing 4T1#31 tumors were intravenously injected with eribulin (Eri) or eribulin-LF (Eri-LF) in combination or not with anti–PD-1 Ab (200 μg/mouse) on a Q7D×2 schedule ( n = 6 per group). Data are presented as mean + SEM. *, P < 0.05; **, P < 0.01; ****, P < 0.0001; and ns, not significant by two-way repeated-measures ANOVA with Tukey multiple-comparisons test using log-transformed values, and statistical results of the difference of TV on the indicated days are shown. B, Waterfall plots across five independent experiments. The best overall response was calculated by comparing the TV change on day t with the baseline TV on day 1 for t ≥ 8. The best percentage changes from baseline that exceeded 200% are shown as 200% in the plots. C, Antitumor activity of eribulin-LF + anti–PD-1 Ab in RAG tumors. Mice bearing RAG tumors were treated with 0.3 mg/kg Eri-LF (intravenously) on a Q7D×3 schedule + anti–PD-1 Ab (200 μg/mouse; intraperitoneally) on a Q3D×7 schedule or with each monotherapy ( n = 10 per group). Data are presented as mean + SEM. *, P < 0.05 and ****, P < 0.0001 by two-way repeated-measures ANOVA with Dunnett multiple-comparisons test.

Article Snippet: 4T1 mouse breast cancer cells (ATCC, catalog no. CRL-2539, RRID:CVCL_0125) were purchased from the ATCC and cultured in RPMI1640 medium supplemented with 10% FBS and penicillin–streptomycin.

Techniques: Activity Assay, Transplantation Assay, Injection, Transformation Assay

Journal: Molecular Cancer Therapeutics

Article Title: Liposome-Encapsulated Eribulin Shows Enhanced Antitumor Activity over Eribulin for Combination Therapy with Anti–PD-1 Antibody

doi: 10.1158/1535-7163.MCT-22-0475

Figure Lengend Snippet: Effect of eribulin-LF + anti–PD-1 Ab on MVD and TILs. A, Scheme of tumor sampling. When the average TV reached 60 to 120 mm 3 , immunocompetent BALB/c mice bearing 4T1#31 tumors were treated with 0.3 mg/kg eribulin or eribulin-LF + anti–PD-1 Ab (200 μg/mouse) or each monotherapy on day 1 (only 1 injection; intravenously). Tumors were collected on day 8 or day 11 from each mouse and used for the following analyses. FCM, flow cytometry. B and C, Eribulin-LF has superior vascular remodeling activity compared with eribulin. Mice bearing 4T1#31 tumors were treated with 0.3 mg/kg eribulin (Eri; intravenously) or eribulin-LF (Eri-LF; intravenously). Tumors were collected on day 11, and IHC analysis was performed. B, Representative IHC images showing CD31 staining. Bar, 100 μm. C, MVD in whole tumor tissues was quantified ( n = 7–8 per group). Data are presented as mean ± SEM. *, P < 0.05; **, P < 0.01; and ****, P < 0.0001 by one-way ANOVA with Tukey multiple-comparisons test. D and E, Eribulin-LF has vascular remodeling activity. Mice bearing 4T1#31 tumors were treated with 0.3 mg/kg eribulin-LF (Eri-LF; intravenously) + anti–PD-1 Ab (200 μg/mouse; intravenously) or each monotherapy. Tumors were collected on day 11, and IHC analysis was performed. D, Representative IHC images showing CD31 staining. Bar, 100 μm. E, MVD in whole tumor tissue was quantified ( n = 8 per group). Data are presented as mean ± SEM. *, P < 0.05 and ***, P < 0.001 by one-way ANOVA with Dunnett multiple-comparisons test. F, Flow cytometric analysis of TILs in 4T1#31 tumors. Mice bearing 4T1#31 tumors were treated with 0.3 mg/kg eribulin (Eri; intravenously) or eribulin-LF (Eri-LF; intravenously). Tumors were collected on day 11, and TILs from dissociated tumor tissue cells were analyzed by flow cytometric analysis ( n = 8 per group). T cells, CD4 + T cells, CD8 + T cells, and NK cells were gated as CD3 + CD11b − cells, CD4 + CD3 + CD11b − cells, CD8 + CD3 + CD11b − cells, and CD49b + CD3 − cells, respectively. Data are shown as box plots with Tukey-style whiskers. *, P < 0.05; **, P < 0.01; ***, P < 0.001; and ****, P < 0.0001 by one-way ANOVA with Tukey multiple-comparisons test. G, Flow cytometric analysis of TILs in 4T1#31 tumors. Mice bearing 4T1#31 tumors were treated with 0.3 mg/kg eribulin-LF (Eri-LF; intravenously) + anti–PD-1 Ab (200 μg/mouse; intravenously) or each monotherapy. Tumors were collected on day 11, and flow cytometric analysis of TILs was performed ( n = 11–12 per group). T cells, CD4 + T cells, CD8 + T cells, and NK cells were gated as CD3 + CD11b − cells, CD4 + CD3 + CD11b − cells, CD8 + CD3 + CD11b − cells, and CD49b + CD3 − cells, respectively. GzmB + CD8 + T cells were gated as GzmB + cells in CD8 + T cells. GzmB + NK cells were gated as GzmB + NKp46 + CD49b + cells. Data are shown as box plots with Tukey-style whiskers. *, P < 0.05; **, P < 0.01; ***, P < 0.001; and ****, P < 0.0001 by one-way ANOVA with Dunnett multiple-comparisons test. H and I, Eribulin-LF increased infiltration of CD8 + cells. Mice bearing 4T1#31 tumors were treated with 0.3 mg/kg eribulin-LF (Eri-LF; intravenously) + anti–PD-1 Ab (200 μg/mouse; intravenously) or each monotherapy. Tumors were collected on day 11 and IHC analysis was performed. H, Representative IHC images showing CD8 staining. Bar, 100 μm. I, CD8 + cell count per unit area of whole tumor tissues was quantified ( n = 8 per group). Data are presented as mean ± SEM. **, P < 0.01 and ***, P < 0.001 by one-way ANOVA with Dunnett multiple-comparisons test.

Article Snippet: 4T1 mouse breast cancer cells (ATCC, catalog no. CRL-2539, RRID:CVCL_0125) were purchased from the ATCC and cultured in RPMI1640 medium supplemented with 10% FBS and penicillin–streptomycin.

Techniques: Sampling, Injection, Flow Cytometry, Activity Assay, Staining, Cell Counting

Journal: Molecular Cancer Therapeutics

Article Title: Liposome-Encapsulated Eribulin Shows Enhanced Antitumor Activity over Eribulin for Combination Therapy with Anti–PD-1 Antibody

doi: 10.1158/1535-7163.MCT-22-0475

Figure Lengend Snippet: Effect of eribulin-LF on immune-related gene expression. Mice bearing 4T1#31 tumors were treated with 0.3 mg/kg eribulin-LF (Eri-LF; intravenously) + anti–PD-1 Ab (200 μg/mouse; intravenously) or each monotherapy ( n = 10 per group). Tumors were collected on day 8, and total RNA was extracted and analyzed by using an nCounter PanCancer Mouse Immune Profiling Panel. A, GSEA. GSEA was performed by using the CERNO algorithm, and P values were calculated by using the CERNO test. We used the immune response category annotations provided by NanoString, and the genes were classified into 30 gene sets based on their annotations. In the figure, the 30 gene-set categories are shown sorted on the basis of the P values for the 0.3 mg/kg Eri-LF group. *, P < 0.05. #adjusted P < 0.05. B, IFNγ signature score was increased by eribulin-LF. IFNγ signature score was calculated using the gene expression data determined by using the nCounter PanCancer Mouse Immune Profiling Panel. Data are presented as mean ± SEM. *, P < 0.05; **, P < 0.01; and ns, not significant by one-way ANOVA with Dunnett multiple-comparisons test.

Article Snippet: 4T1 mouse breast cancer cells (ATCC, catalog no. CRL-2539, RRID:CVCL_0125) were purchased from the ATCC and cultured in RPMI1640 medium supplemented with 10% FBS and penicillin–streptomycin.

Techniques: Gene Expression

Journal: Molecular Cancer Therapeutics

Article Title: Liposome-Encapsulated Eribulin Shows Enhanced Antitumor Activity over Eribulin for Combination Therapy with Anti–PD-1 Antibody

doi: 10.1158/1535-7163.MCT-22-0475

Figure Lengend Snippet: Correlation between infiltration of CD8 + cells into tumor and vascular remodeling by eribulin-LF. Representative IHC images of CD8/CD31 double staining. Mice bearing 4T1#31 tumors were treated with 0.3 mg/kg eribulin-LF (Eri-LF; intravenously) + anti–PD-1 Ab (200 μg/mouse; intravenously) or each monotherapy. Tumors were collected on day 11, and IHC analysis was performed. Tissues were stained with anti-CD8α (green) and anti-CD31 (red) Abs, and nuclei were counterstained with DAPI (blue). The dashed line marked “C” indicates the tumor core. Bar, 100 μm.

Article Snippet: 4T1 mouse breast cancer cells (ATCC, catalog no. CRL-2539, RRID:CVCL_0125) were purchased from the ATCC and cultured in RPMI1640 medium supplemented with 10% FBS and penicillin–streptomycin.

Techniques: Double Staining, Staining

Journal: Molecular Cancer Therapeutics

Article Title: Liposome-Encapsulated Eribulin Shows Enhanced Antitumor Activity over Eribulin for Combination Therapy with Anti–PD-1 Antibody

doi: 10.1158/1535-7163.MCT-22-0475

Figure Lengend Snippet: Effect of eribulin-LF on expression of ICAM-1 in vascular ECs. Eribulin-LF induced ICAM-1 expression in vascular ECs in 4T1#31 tumors. Mice bearing 4T1#31 tumors were treated with the indicated dose of eribulin-LF (Eri-LF; intravenously). Tumors were collected on day 8, and CD31 + vascular ECs were enriched by magnetic-activated cell sorting. After enrichment of CD31 + vascular ECs, the expression level of ICAM-1 on vascular ECs was evaluated by flow cytometric analysis. A, Representative dot plots (CD45 vs. CD31). Cell suspensions obtained from tumor tissues (before enrichment) and EC-enriched cell suspensions after magnetic-activated cell sorting (after enrichment) were analyzed by flow cytometric analysis. Vascular ECs were gated as CD31 + CD45 − cells (blue lines). B, Representative histogram of ICAM-1 expression in the population gated on CD31 + vascular ECs. The gray line represents the isotype control; and the black, blue, and red lines represent ICAM-1 in the nontreatment, 0.3 mg/kg Eri-LF, and 1.0 mg/kg Eri-LF groups, respectively. C, Mean fluorescence intensities ( n = 5 per group). Data are presented as mean ± SEM. ***, P < 0.001 and ****, P < 0.0001 by one-way ANOVA with Dunnett multiple-comparisons test.

Article Snippet: 4T1 mouse breast cancer cells (ATCC, catalog no. CRL-2539, RRID:CVCL_0125) were purchased from the ATCC and cultured in RPMI1640 medium supplemented with 10% FBS and penicillin–streptomycin.

Techniques: Expressing, FACS, Control, Fluorescence