Journal: Journal of the American Chemical Society

Article Title: Encapsulation of Gold-Based Anticancer Agents in Protease-Degradable Peptide Nanofilaments Enhances Their Potency

doi: 10.1021/jacs.2c09820

Figure Lengend Snippet: Cell Viability of gold anticancer agents, gold-loaded peptide nanofilaments ( PD+1 , PD+2 , AD+1 , AD+2 ), and peptides ( PD , AD ) in three human cell lines. Caki-1 (renal cancer), MDA-MB-231 (Triple Negative Breast Cancer), and IMR-90 (lung fibroblasts) incubated with drug-loaded peptide for 72 h. A) Compound 1 , B) Compound 2 . (* represents P <0.05, ** represents P <0.01, *** represents P <0.001).

Article Snippet: Human breast adenocarcinoma line MDA-MB-231 and human lung fibroblast line IMR-90 were obtained from the American Type Culture Collection (ATCC; Manassas, Virginia, USA) and cultured with Dulbecco’s modified Eagle’s medium (DMEM; Fisher Scientific) containing 10 % fetal bovine serum, certified, heat inactivated, US origin (FBS; Fisher Scientific), 1 % minimum essential media (MEM) nonessential amino acids (NEAA; Fisher Scientific), and 1 % penicillin–streptomycin (PenStrep; Fisher Scientific).

Techniques: Incubation

Journal: NPJ precision oncology

Article Title: Chimeric antibody targeting unique epitope on onco-mucin16 reduces tumor burden in pancreatic and lung malignancies.

doi: 10.1038/s41698-023-00423-7

Figure Lengend Snippet: Fig. 4 ch5E6 treatment leads to a substantial decrease in the growth of organoids derived from PC patients or pancreatic and lung cancer genetically engineered mouse models. a PC patient and b KPC mouse organoid staining with ch5E6 indicating MUC16 expression (green) and specific binding of mAb compared to no binding with isotype control mAb huIgG1. The binding of ch5E6 to the ductal cells (yellow) was illustrated by colocalization with CK-19 (red) staining in human and mouse pancreatic tumor organoids. Scale bar 10 µm.The representative images for ch5E6 treated. c Human PC patient. d KPC and (e) KPA mouse-derived organoids obtained by real-time kinetics using Incucyte live cell imaging system. The data were quantitated for organoid counts using essence incucyte software, plotted as a graph of change in organoid counts or area over time (3–5 days) for both ch5E6 and huIgG1 treatments, and is shown in parallel. Error bars indicate SEM. *P < 0.05; **P < 0.01.

Article Snippet: We performed Real time MT-glo assay to determine the effect of anti-MUC16 chimeric mAb5E6 (ch5E6) and isotype control mAb (huIgG1; Bio X Cell).

Techniques: Derivative Assay, Staining, Expressing, Binding Assay, Control, Live Cell Imaging, Software

Journal: NPJ precision oncology

Article Title: Chimeric antibody targeting unique epitope on onco-mucin16 reduces tumor burden in pancreatic and lung malignancies.

doi: 10.1038/s41698-023-00423-7

Figure Lengend Snippet: Fig. 6 Inhibition of EMT by ch5E6 is validated in PC and NSCLC cell line-derived xenografts. a Immunofluorescence analysis showing a decrease in pFAK(Y397) and N-cadherin expression in xenograft tumors of SW1990 cells treated with ch5E6 compared to isotype control mAb huIgG1 group (n = 6–8 fields/tissue: three animals). The data was plotted for changes in fluorescence intensity using GraphPad Prism 9 and is shown in parallel. Nuclei were stained with DAPI. b Immunofluorescence analysis of ch5E6 treated SW1573 cell line-derived xenografts showing a reduction in pFAK(Y397) and N-cadherin levels compared to isotype control mAb huIgG1 group (n = 6–8 fields/tissue: 3 animals). Scale bar, 10 µm; magnified images, 2 µm. No significant changes in the intensity of MUC16 were seen in the ch5E6 treated versus isotype control tumors derived from both cancers. c, d Immunoblot analysis of ch5E6 treated PDAC and NSCLC tumor lysates showing a substantial decrease in phosphorylated levels of FAK(Y397), p70S6K(T389) and N-cadherin as compared to huIgG1 treatment. Error bars indicate SEM. Scale bar, 400 μm; magnified images, 100 μm; *P < 0.05; **P < 0.01.

Article Snippet: We performed Real time MT-glo assay to determine the effect of anti-MUC16 chimeric mAb5E6 (ch5E6) and isotype control mAb (huIgG1; Bio X Cell).

Techniques: Inhibition, Derivative Assay, Expressing, Control, Staining, Western Blot

Journal: NPJ precision oncology

Article Title: Chimeric antibody targeting unique epitope on onco-mucin16 reduces tumor burden in pancreatic and lung malignancies.

doi: 10.1038/s41698-023-00423-7

Figure Lengend Snippet: Fig. 7 MUC16 and N-cadherin are clinically correlated in patient tumors. a, b Representative images and quantitative illustration of immunohistochemical analyses demonstrate a strong positive correlation between MUC16 and N-cadherin (R = 0.84) in both primary PC tumors (n = 10) and liver metastasis (R = 0.99) samples (n = 8). Scale bar, 400 µm; magnified images, 100 µm. *P < 0.05; **P < 0.01. c Representative images of immunofluorescence analysis showing coexpression of MUC16 (green) and N-cadherin (red) in primary PDAC tumors compared to no MUC16 and N-cadherin in normal pancreatic sections. Scale bar, 20 µm; magnified images, 5 µm. d Schematic diagram representing ch5E6 induced downregulation of MUC16 mediated EMT resulting in its anti-tumor potential in PC and NSCLC. Overall, anti- MUC16 chimeric mAb5E6 (ch5E6) binds to the cell surface-tethered domain of MUC16, interferes with oncogenic pFAK/p70S6K/N-cadherin signaling associated with MUC16-mediated EMT, and reduces tumor burden in both PC and NSCLC models.

Article Snippet: We performed Real time MT-glo assay to determine the effect of anti-MUC16 chimeric mAb5E6 (ch5E6) and isotype control mAb (huIgG1; Bio X Cell).

Techniques: Immunohistochemical staining

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Therapeutic impact of Nintedanib with paclitaxel and/or a PD-L1 antibody in preclinical models of orthotopic primary or metastatic triple negative breast cancer

doi: 10.1186/s13046-018-0999-5

Figure Lengend Snippet: Paclitaxel and its combination with nintedanib increased median survival in the advanced metastatic breast cancer LM2–4 model. a ) Kaplan-Meier survival curves and median survival values. Paclitaxel (PTX) significantly increased median survival compared to the control group ( p = 0.033; Log-rank (Mantel Cox) Test, n = 8–10). Combination therapy increased median survival (81 days vs 60.5 days, control group) but it did not reach significance. Treatment started around 40 days after cell implantation. b) Effect of sunitinib alone and when combined with PTX in the advanced metastatic LM2–4 breast cancer model. Kaplan-Meier survival curves and median survival values. Modified from Guerin et al., 2013 . PTX alone increased survival whereas sunitinib alone did not, and adding sunitinib to PTX did not result in increased efficacy

Article Snippet: The EMT-6 (ATCC® CRL-2755TM) mouse breast cancer cell line and the derived variant EMT-6/CDDP -selected in vivo for acquired resistance to cisplatin [ ]-, were cultured in DMEM medium with 5% FBS at 37 °C in 5% CO 2.

Techniques: Control, Modification

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Therapeutic impact of Nintedanib with paclitaxel and/or a PD-L1 antibody in preclinical models of orthotopic primary or metastatic triple negative breast cancer

doi: 10.1186/s13046-018-0999-5

Figure Lengend Snippet: Improvement of immunotherapy efficacy when treating primary tumors with nintedanib combination therapy . a ) Tumor growth in the primary EMT-6 breast cancer model. Treatment was started when average tumor volume was 120mm 3 , around 7 days after cell implantation. Statistical analysis on day 27. ANOVA followed by Tukey’s Multiple Comparison Test * p < 0.05, ** p < 0.01. Data are presented as means ± SD, n = 6. Region of flat line in the curves means that tumor in remaining mice had regressed, and in the case of mice treated with PD-L1 antibody, tumors grew back. Mice were treated with nintedanib and/or paclitaxel (PTX) for 70 days, and then treatment stopped. b) Tumor growth in the primary EMT-6/CDDP model. Treatment was started when average tumor volume was 120mm 3 , 7 days after cell implantation. Statistical analysis on day 27. Kruskal-Wallis test followed by Dunn’s Multiple Comparison Test, ** p < 0.01. Data are presented as means ± SD, n = 9–12. c-f) Effect of nintedanib, paclitaxel, anti-PD-L1 and the drug combinations on c) Vascularity; d) Proliferation; e) CD8+ tumor infiltrating cells; and f) Level of Necrosis. Histology and immunohistochemistry analyses were performed on tumor samples obtained from mice implanted with EMT-6/CDDP cells. Treatment was started when average tumor volume was 120mm 3 and all mice were sacrificed after 10 days of treatment. The Mann-Whitney test was used for statistical analyses. Data are presented as means ± SD, n = 6–7

Article Snippet: The EMT-6 (ATCC® CRL-2755TM) mouse breast cancer cell line and the derived variant EMT-6/CDDP -selected in vivo for acquired resistance to cisplatin [ ]-, were cultured in DMEM medium with 5% FBS at 37 °C in 5% CO 2.

Techniques: Comparison, Immunohistochemistry, MANN-WHITNEY

Journal: Cells

Article Title: Chronic High-Salt Diet Activates Tumor-Initiating Stem Cells Leading to Breast Cancer Proliferation

doi: 10.3390/cells13110912

Figure Lengend Snippet: Enumeration of TISC frequency in various murine passages following dietary salt modification. The frequency of CD44+CD24−TISCs cells in the isolated tumor cells among RS (grey) and HS (red) diet cohorts was enumerated by flow cytometry. ( A , B ) Representative flow cytometry plot of TISCs from passage 1 and passage 4 of Py230-C57Bl/6J HS diet cohort. ( C , D ) Changes in TISC frequency with each passage in Py230-C57Bl/6J ( C ) and 4T1-BALB/cJ ( D ) murine tumor models. ( E – N ) The mRNA expression of TISC markers Cadherin 1 ( E , J ), Snail2 ( F , K ), Aldh1A1 ( G , L ), Sox2 ( H , M ), and ITGA6 ( I , N ) in four passages of Py230-C57Bl/6J and 4T1-BALB/cJ tumor models (respectively). Data analyzed by one-way ANOVA for multiple comparisons and presented as mean ± SEM; n = 8 (biological replicates) per cohort; (*) p -value < 0.05.

Article Snippet: The TISCs were isolated from murine tumors by CD24 − CD44 + breast tumor stem cells using immunomagnetic beads as per the manufacturer’s specifications and reagents (catalog#MAGH111, R&D Systems, Minneapolis, MN, USA).

Techniques: Modification, Isolation, Flow Cytometry, Expressing

Journal: Cells

Article Title: Chronic High-Salt Diet Activates Tumor-Initiating Stem Cells Leading to Breast Cancer Proliferation

doi: 10.3390/cells13110912

Figure Lengend Snippet: TGFβ signaling induced the expression of immune-inhibitory CD80 on TISCs. ( A ) Representative flow cytometry histogram of TGFβR2 expression in passages 1 (blue), 2 (pink), 3 (olive green), and 4 (purple) from the HS diet cohort of the Py230-C57Bl/6J tumor model. ( B ) Comparison of the relative expression of TGFβR2 in each of the four passages on CD4+CD24-TISCs isolated from RS (grey) and HS (red) diet cohorts of the Py230-C57Bl/6J tumor model. ( C ) Representative flow cytometry histogram of pSMAD2/3 intracellular expression in each of the four passages from the HS diet cohort of the Py230-C57Bl/6J tumor model. ( D ) Comparison of the relative expression of pSMAD2/3 in each of the four passages on CD4+CD24-TISCs isolated from the RS and HS diet cohorts of the Py230-C57Bl/6J tumor model. ( E ) Representative flow cytometry histogram of CD80 surface expression in each of the four passages from the HS diet cohort of the Py230-C57Bl/6J tumor model. ( F ) Comparison of the relative expression of CD80 in each of the four passages on CD4+CD24-TISCs isolated from the RS and HS diet cohorts of the Py230-C57Bl/6J tumor model. ( G ) Representative flow cytometry plot to determine the TGFβR2/CD80 double-positive TISCs in passage 4 of the HS diet cohort from the Py230-C57Bl/6J tumor model. ( H ) Comparison of the relative expression of TGFβR2/CD80 double-positive TISCs in each of the four passages on CD4+CD24-TISCs isolated from the RS and HS diet cohorts of the Py230-C57Bl/6J tumor model. ( I ) Comparison of the relative expression of CD80 following TGFβ (80 ng/mL) stimulation for 5 days on CD4+CD24-TISCs isolated from each of the four passages of the RS and HS diet cohorts in the Py230-C57Bl/6J tumor model. ( J – M ) Comparison of the relative expression of TGFβR2 ( J ), pSMAd2/3 ( K ), CD80 ( L ), and TGFβR2/CD80 double-positive cells ( M ) in each of the four passages on CD4+CD24-TISCs isolated from the RS (grey) and HS (red) diet cohorts of the 4T1-BALB/cJ tumor model. ( N ) Comparison of the relative expression of CD80 following TGFβ (80 ng/mL) treatment for 5 days on CD4+CD24-TISCs isolated from each of the four passages of the RS and HS diet cohorts of the 4T1-BALB/cJ tumor model. Data analyzed by one-way ANOVA for multiple comparisons and presented as mean ± SEM; n = 8 (biological replicates) per cohort; (*) p -value < 0.05.

Article Snippet: The TISCs were isolated from murine tumors by CD24 − CD44 + breast tumor stem cells using immunomagnetic beads as per the manufacturer’s specifications and reagents (catalog#MAGH111, R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Flow Cytometry, Comparison, Isolation