Journal: Aiche Journal. American Institute of Chemical Engineers

Article Title: Metastatic cancer cell attachment to endothelium is promoted by endothelial glycocalyx sialic acid degradation

doi: 10.1002/aic.16634

Figure Lengend Snippet: Preliminary data was collected regarding human ECs (HUVEC), their GCX, and the extent of their recruitment of 4T1 breast cancer cells in comparison with RFPEC controls. This human EC data confirms and validates the rat EC data that was the focus of this report. (a) Phase image shows that an untreated HUVEC layer is healthy. (b) WGA‐labeled untreated HUVEC reveals the presence of GCX even in the absence of physiological flow stimulation, which is usually required for in vitro human EC studies. (c) Low expression of HS is observed when these HUVEC, which lack flow stimulation, are labeled with HS antibody. The limited HS is presumably insufficient to expose the EC surface adhesion molecules to 4T1 breast cancer cells, because WGA is abundant enough to compensate and provide adequate coverage. (d) As expected, the level of attachment of 4T1 breast cancer cells to untreated HUVEC is low, similar to what was observed in untreated RFPEC. (e) Phase image shows healthy untreated RFPEC monolayer. (f) Fluorescent image of WGA‐labeled untreated RFPEC monolayer shows intact GCX. (g) Expression of HS is abundant in RFPEC without flow stimulation. (h) Picture shows attachment of 4T1 breast cancer cells to RFPEC monolayers. (i) Plot shows that the difference between RFPEC and HUVEC adhesiveness to 4T1 breast cancer cells is statistically not significant (ns). N = 3 for both cell types that were studied. EC, endothelial cell; GCX, glycocalyx; HS, heparan sulfate; RFPEC, rat fat pad endothelial cell; WGA, wheat germ agglutinin

Article Snippet: In brief, Stage IV metastatic mouse breast cancer cells (4T1) (ATCC) at passages 5 to 10 were cultured in DMEM/1% PS/10% FBS, humidity, 37°C, and 5% CO 2 .

Techniques: Comparison, Labeling, In Vitro, Expressing

Journal: Science Advances

Article Title: Integrative analysis of mRNA stability regulation uncovers a metastasis-suppressive program in breast cancer

doi: 10.1126/sciadv.aea9061

Figure Lengend Snippet: ( A to C ) Lung colonization assay for control and either RBMS3 knockdown or RBMS3 overexpression. Luciferase-labeled cells were injected via tail vein, and their metastatic growth in the lungs was measured over time. The magnitude of the signal, shown here as a heatmap for a representative mouse from each cohort, reflects the metastatic burden. Bars on time-course plots show means ± SEM, and ANOVA was performed (left). Also included are area under the curve (AUC) of log-normalized signal in the lungs of mice over 25 days. Summary overlay shows means ± SEM and Mann-Whitney U test used to determine statistical significance (middle). Also, hematoxylin and eosin–stained lung sections for exemplary mice (right). (A) Lung colonization assays of control ( N = 4 biological replicates) and RBMS3 knockdown MDA-MB-231 cells ( N = 4 biological replicates). (B) Lung colonization assays for control ( N = 5 biological replicates) and RBMS3 overexpression MDA-LM2 cells ( N = 5 biological replicates). (C) Lung colonization assays for control ( N = 5 biological replicates) and RBMS3 knockdown HCC1806 cells ( N = 5 biological replicates). ( D ) Invasion assays for control and RBMS3 knockdown MDA-MB-231 cells. Images are representative (i.e., median) in each group. In processed images, black is empty space and white is cells. The graph shows fraction that is white (i.e., % area covered), and the Mann-Whitney U test was used for statistical comparison. * P < 0.05; *** P < 0.001.

Article Snippet: We measured TXNIP expression using qPCR in 96 clinical samples across all stages of breast cancer, namely 5 normal epithelial, 23 stage I, 30 stage II, 29 stage III, and 9 stage IV metastatic biopsies (Origene, BCRT102, BCRT103), from which 90 samples yielded sufficient amount of cDNA.

Techniques: Control, Knockdown, Over Expression, Luciferase, Labeling, Injection, MANN-WHITNEY, Staining, Comparison

Journal: Science Advances

Article Title: Integrative analysis of mRNA stability regulation uncovers a metastasis-suppressive program in breast cancer

doi: 10.1126/sciadv.aea9061

Figure Lengend Snippet: ( A ) Schematic of the dual-guide CRISPR interference (CRISPRi) screen. ( B ) Analysis of CRISPRi screen comparing the abundance of cells expressing each guide between in vivo– and in vitro–grown cells with DESeq2. TXNIP was observed to be present at high levels in vivo and low levels in vitro indicating high metastasis but low proliferation. ( C ) Comparative analysis of TXNIP to RBMS3 expression in the METABRIC cohort . Shown is Pearson correlation with t statistic for nonzero correlation. ( D ) Analysis of disease-free survival in the METABRIC cohort relative to TXNIP expression. ( E ) Meta-analysis of relapse-free survival in smaller published cohorts relative to TXNIP expression. (D and E) The Mantel-Cox test was used to measure significance. Low TXNIP expression is indicative of poor prognosis for disease-free survival in patients with breast cancer. ( F ) The expression levels of TXNIP in 90 tumor samples at different stages of breast cancer was measured by qPCR; bars show means and SEM. ANOVA was performed. ( G ) The expression levels of TXNIP in RBMS3 knockdown MDA-MB-231 cells and control cells was measured by qPCR and compared using Mann-Whitney U test. ( H ) Lung colonization assays of control ( N = 5 mice), TXNIP knockdown ( N = 5 mice), and RBMS3-TXNIP double knockdown ( N = 5 mice) in MDA-MB-231 cells. Luciferase-labeled cells were injected via tail vein, and their metastatic growth in the lungs was measured over time. ANOVA was performed; bars indicate means and SEM. ( I ) Molecular mechanism of RBMS3 metastasis suppression through posttranscriptional regulatory action.

Article Snippet: We measured TXNIP expression using qPCR in 96 clinical samples across all stages of breast cancer, namely 5 normal epithelial, 23 stage I, 30 stage II, 29 stage III, and 9 stage IV metastatic biopsies (Origene, BCRT102, BCRT103), from which 90 samples yielded sufficient amount of cDNA.

Techniques: CRISPR, Expressing, In Vivo, In Vitro, Knockdown, Control, MANN-WHITNEY, Luciferase, Labeling, Injection

Journal: Cancer immunology, immunotherapy : CII

Article Title: Intralesional treatment of metastatic melanoma: a review of therapeutic options

doi: 10.1007/s00262-016-1952-0

Figure Lengend Snippet: Clinical trials with novel intralesional agents

Article Snippet: Agent Sponsor Phase Indication Ref. N/A OncoVEX GM-CSF Biovex I breast, gastrointestinal adenocarcinoma, malignant melanoma, HNSCC 37 NCT00289016 OncoVEX GM-CSF Biovex II Stage IIIC and IV melanoma 38 NCT00769704 Talimogene laherparepvec Amgen III Unresected stage IIIB/C and IV melanoma 8 NCT01740297 T-Vec + Ipilimumab Amgen Ib/II Unresected stage IIIB/C and IV melanoma 42 NCT02263508 T-Vec + Pembrolizumab Amgen, MSD III Unresectable Stage IIIB to IVM1c melanoma 43 NCT02211131 T-Vec Amgen II Resectable Stage IIIB/C and IVM1a melanoma (Neoadjuvant) N/A NCT00219843 PV-10 (rose bengal) Provectus I Stage III and IV metastatic melanoma 45 NCT00521053 PV-10 (rose bengal) Provectus II Stage III and IV metastatic melanoma 47 NCT02288897 PV-10 (rose bengal) Provectus III Stage IIIB, IIIC or Stage IV M1a with no active nodal metastases N/A NCT01253096 Darleukin (L19IL2) Philogen II Stage IIIB/C melanoma 15 NCT02076633 Daromun (L19IL2 + L19TNF) Philogen II Stage IIIB/C and IVM1a melanoma 56 NCT02938299 Daromun (L19IL2 + L19TNF) Philogen III Resectable Stage IIIB/C melanoma (Neoadjuvant) N/A Open in a separate window N/A = Not applicable caption a8 Clinical trials with novel intralesional agents

Techniques: