Journal: Oncotarget

Article Title: A transcriptomic signature mediated by HOXA9 promotes human glioblastoma initiation, aggressiveness and resistance to temozolomide

doi:

Figure Lengend Snippet: (A) qPCR confirming the overexpression of HOXA9 in hTERT/E6/E7-HOXA9 and U87MG-HOXA9 cells and HOXA9 -silencing in U251-shHOXA9 and GBML18-shHOXA9 cells, comparing to their respective control counterparts. (B) Venn diagram summarizing the number of differentially expressed transcripts in the microarray data in all cell lines. The numbers in each area represent the total number of transcripts within each intersection. (C–F) DAVID was used to query the HOXA9-transcriptome from each cell line (C, hTERT/E6/E7; D, U87MG; E, U251; F, GBML18), in order to identify enriched biological terms on the differentially expressed genes extracted from the microarray data. Statistically significant enriched GO terms are shown for each cell line.

Article Snippet: For HOXA9 silencing , GBML18 and U251 cells were transfected with pGFP-V-RS plasmid containing HOXA9-specific shRNAs or non-effective shRNAs sequences (Origene).

Techniques: Over Expression, Microarray

Journal: Oncotarget

Article Title: A transcriptomic signature mediated by HOXA9 promotes human glioblastoma initiation, aggressiveness and resistance to temozolomide

doi:

Figure Lengend Snippet: (A–D) GSEA reveals that the HOXA9 transcriptomes in hTERT/E6/E7 (A) and U251 (C) cells are associated with transcriptional signatures of glioma stem-like cells (Enrichment Score, ES = −0.54, False Discovery Rate, FDR = 0.19; and ES = 0.50, FDR = 0.11, respectively); in U87MG cells (B) , the HOXA9 transcriptome is positively associated with genes that are upregulated in embryonic stem cells (ES = 0.50, FDR < 0.0001); in GBML18 cells (D) , HOXA9 transcriptome is inversely associated with genes upregulated during the neuronal differentiation (ES = 0.75, FDR = 0.21). (E) Representative phase contrast photographs of hTERT/E6/E7 and U87MG neurospheres are shown. (F) Quantification of neurospheres number and size for each cell line ( n = 3; * p < 0.05; *** p < 0.001). (G) Immunofluorescence showing increased Nestin staining in HOXA9-positive cells.

Article Snippet: For HOXA9 silencing , GBML18 and U251 cells were transfected with pGFP-V-RS plasmid containing HOXA9-specific shRNAs or non-effective shRNAs sequences (Origene).

Techniques: Immunofluorescence, Staining

Journal: Oncotarget

Article Title: A transcriptomic signature mediated by HOXA9 promotes human glioblastoma initiation, aggressiveness and resistance to temozolomide

doi:

Figure Lengend Snippet: (A and B) Determination of the half inhibitory concentration (IC 50 ) values after 6 days of temozolomide (TMZ) treatment in HOXA9 -positive or HOXA9 -negative hTERT/E6/E7 and U87MG (A), and HOXA9 -silenced or control U251 and GBML18 (B) cell lines. (C–F) Cell viability trypan blue assays in HOXA9 -negative/low or HOXA9 –positive/high hTERT/E6/E7 (C), U87MG (D), U251 (E) and GBML18 (F) cells, exposed to temozolomide or vehicle. (G) Cell death was evaluated by annexin V staining followed by flow cytometry in HOXA9 -positive/high and HOXA9 -negative/low hTERT/E6/E7, U87MG, U251 and GBML18 cell lines, both in basal conditions and after exposure to temozolomide (TMZ). HOXA9 expression decreases cell death of all GBM cell models, both in basal conditions and after TMZ treatment, except in basal conditions for U251 cell line. (H) Cell invasion in the same cell lines, both in basal conditions and after exposure to TMZ. HOXA9 increases the invasion of hTERT/E6/E7, U87MG, and GBML18 cells. U251 cells did not show significant differences in invasion profiles due to HOXA9 levels or TMZ treatment. Results are representative of at least three independent experiments, performed in triplicates (data points represent mean ± SEM). Statistical differences were calculated by Student's t -test (panels A, B, G, H) and two-way ANOVA (panels C–F) (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: For HOXA9 silencing , GBML18 and U251 cells were transfected with pGFP-V-RS plasmid containing HOXA9-specific shRNAs or non-effective shRNAs sequences (Origene).

Techniques: Concentration Assay, Staining, Flow Cytometry, Expressing

Journal: Journal of Biomedical Optics

Article Title: Prediction of optimal contrast times post-imaging agent administration to inform personalized fluorescence-guided surgery

doi: 10.1117/1.JBO.25.11.116005

Figure Lengend Snippet: Targeted and control imaging agents in three different imaging agent classes: peptides, affibodies, and antibodies. Mouse human cancer xenograft (subcutaneous thigh tumor models) fluorescent imaging dynamics from the peptide-based imaging agent (targeted: IRDye ® 800CW EGF, control: IRDye ® 700DX) studies are presented from the moderate EGFR-expressing cell line (human glioblastoma; U251); the high EGFR-expressing cell line (human epidermoid, A431); the affibody-based imaging agent study in U251 xenografts (targeted: IRDye ® 800CW anti-EGFR affibody, control: IRDye ® 680RD negative control affibody ® ); the antibody-based imaging agent study in U251 xenografts (targeted: IRDye ® 800CW Cetuximab, control: IRDye ® 700DX IgG). (a)–(d) The mean fluorescence measured for the targeted and control imaging agents in the tumors in each group (errors are SD between animals). (e)–(h) The same information in a proportionally sized region-of-interest in the muscle surrounding the tumors in each case.

Article Snippet: The mean AUROCs of the affibody U251 group were 0.67 ± 0.09 and 0.8 ± 0.1 in the SA and PA groups, respectively.

Techniques: Control, Imaging, Expressing, Negative Control, Fluorescence

Journal: Journal of Biomedical Optics

Article Title: Prediction of optimal contrast times post-imaging agent administration to inform personalized fluorescence-guided surgery

doi: 10.1117/1.JBO.25.11.116005

Figure Lengend Snippet: Comparison of the PA and SA FGS protocols. The AUROCs and the CVRs measured from PA (red data) and SA (blue data) analyses of the experimental results as a function of time post-imaging-agent injection. The mean of the AUROCs measured in each group (errors are SD between animals) for (a) the U251-peptide group, (b) the A431-peptide group, (c) the U251-affibody group, and (d) the U251-antibody group. The CVR measured in each group (errors are SD between animals) for (e) the U251-peptide group, (f) the A431-peptide group, (g) the U251-affibody group, and (h) the U251-antibody group. The third column depicts maps of targeted imaging agent fluorescence the SA (targeted; green) and the PA binding potentials ( B P ratio ; grayscale) for three randomly selected mice in each of (i) the U251-peptide group, (j) the A431-peptide group, (k) the U251-affibody group, and (l) the U251-antibody group. The yellow dashed circles depict the location of the tumors and the red dashed circles show the normal muscle tissue surrounding the tumor that were selected as representative of “background.” The ROIs were selected based on the white-light images (not shown).

Article Snippet: The mean AUROCs of the affibody U251 group were 0.67 ± 0.09 and 0.8 ± 0.1 in the SA and PA groups, respectively.

Techniques: Comparison, Imaging, Injection, Fluorescence, Binding Assay

Journal: Journal of Biomedical Optics

Article Title: Prediction of optimal contrast times post-imaging agent administration to inform personalized fluorescence-guided surgery

doi: 10.1117/1.JBO.25.11.116005

Figure Lengend Snippet: Simulation results for three different classes of imaging agents: peptides, affibodies and antibodies. Simulated fluorescence signal are presented for mouse human cancer xenograft (subcutaneous thigh tumor models) fluorescent imaging dynamics from the peptide-based imaging agent (targeted: IRDye ® 800CW EGF, control: IRDye ® 700DX) from the moderate EGFR-expressing cell line (human glioblastoma; U251); the high EGFR-expressing cell line (human epidermoid, A431); the affibody-based imaging agent study in U251 xenografts (targeted: IRDye ® 800CW anti-EGFR affibody, control: IRDye ® 680RD negative control affibody ® ); and the antibody-based imaging agent study in U251 xenografts (targeted: IRDye ® 800CW Cetuximab, control: IRDye ® 700DX IgG). (a)–(d) The mean fluorescence measured for the targeted and control imaging agents in the tumors in each group (errors are SD between animals). (e)–(h) The same information in a proportionally sized region-of-interest in the muscle surrounding the tumors in each case.

Article Snippet: The mean AUROCs of the affibody U251 group were 0.67 ± 0.09 and 0.8 ± 0.1 in the SA and PA groups, respectively.

Techniques: Imaging, Fluorescence, Control, Expressing, Negative Control

Journal: Journal of Biomedical Optics

Article Title: Prediction of optimal contrast times post-imaging agent administration to inform personalized fluorescence-guided surgery

doi: 10.1117/1.JBO.25.11.116005

Figure Lengend Snippet: Simulation results for three different classes of imaging agents: peptides, affibodies and antibodies. The mean ± SD of AUROC (blue = SA, red = PA) for (a) the U251-peptide, (b) the A431-peptide, (c) the U251-affibody and (d) the U251-antibody simulated groups. The mean ± SD of CVR (blue = SA, red = PA) for (e) the U251-peptide, (f) the A431-peptide, (g) the U251-affibody, and (h) the U251-antibody simulated groups.

Article Snippet: The mean AUROCs of the affibody U251 group were 0.67 ± 0.09 and 0.8 ± 0.1 in the SA and PA groups, respectively.

Techniques: Imaging