Journal: Neoplasia (New York, N.Y.)

Article Title: Twist1-induced suppression of oncogene-induced senescence in non-small cell lung cancer requires the transactivation domain of Twist1

doi: 10.1016/j.neo.2025.101179

Figure Lengend Snippet: The F191G mutation in the transactivation domain of Twist1 abrogates Twist1-induced acceleration of tumorigenesis and aggressiveness. ( A ) Genetically engineered mouse model for CR, CRT and CRF191G expressing Kras G12D with or without Twist1 wt or Twist1 F191G transactivation-deficient mutant in the mouse lung epithelium under doxycycline treatment. Transgenes are: CCSP-rtTA (C), tetO7- KrasG12D (R), Twist1 wt -tetO 7 -luc (T), and Twist1 F191G -tetO 7 -luc (F191G). ( B ) IHC for TWIST1 expression in lung tissue sections of CR, CRT and CRF191G (magnification 20x) from week 4 to 50 on doxycycline treatment. Intensity quantifications in supplementary figure S1E. ( C ) Kaplan-Meier curves for Tumor-Free Survival (log-rank (Mantel-Cox) analysis: P > 0.05(ns)/ P < 0.05(*)/0.01(**)/0.001(***)), and Tumor-Free Survival median for the CR, CRT and CRF191G. Tumor positivity was determined by CT imaging. ( D ) H&E staining (magnification 4x) of mouse lung tissue sections, number of nodules, and percentages of subjects positive for adenocarcinoma development in CR, CRT and CRF191G. Data are presented as average values ± S.D. ( n = 3-6 animal subjects per time point).

Article Snippet: Twist1 wt and Twist1 F191G exon 1 of founders were sequenced to validate the model by the Johns Hopkins Medicine Genetic Resources Core Facility, Baltimore, MD, USA ( ).

Techniques: Mutagenesis, Expressing, Imaging, Staining

Journal: Neoplasia (New York, N.Y.)

Article Title: Twist1-induced suppression of oncogene-induced senescence in non-small cell lung cancer requires the transactivation domain of Twist1

doi: 10.1016/j.neo.2025.101179

Figure Lengend Snippet: The transactivation function of TWIST1 sustains pro-tumorigenic and pro-invasive transcriptional programs in HBEC. ( A ) In vitro HBEC model overexpressing or not human TWIST1 wt or human TWIST F187G transactivation-deficient mutant and modified or not to overexpress the oncogene HRas G12V (adenovector HRas G12V -EGFP) or control (adenovector EGFP). Tests were done at 72-hours post-adenoviral transduction for HRas G12V -EGFP or control EGFP in HBEC Parental, HBEC— Control , HBEC- TWIST1 wt and HBEC- TWIST F187G . ( B ) Western-blot for TWIST1, HRas G12V and p53 (actin served as loading control). (Average fold change for Twist1 wt or Twist1 F187G expression analyzed by signal intensity quantification (Fiji-ImageJ) were respectively increased by x59.8 and x52.8 compared to HBEC— Control condition); ( C-E ) RNA sequencing analyses on HBEC models in vitro ( n = 3): ( C ) Overlap of DEGs for each HBEC model under HRas G12V compared to HCE condition; ( D ) Heatmap of differentially expressed genes dysregulated in HTH condition. Color gradient from blue (down-regulated) to purple (up-regulated) illustrates the scaled fold-change for each gene in each comparison (see also Fig. S3C); ( E ) Dot plot illustrating the gene set enrichment of cancer-related pathways using the DEGs for each comparison. Color gradient of the inner circle from green to purple represents the normalized enrichment score and the color gradient for the outer ring from grey to red represents the significance scaled as -log10(p-value). [DEGs: differentially expressed genes; HCE: HBEC co-expressing vector control and vector EGFP; HCH: HBEC co-expressing vector control and HRas G12V -EGFP; HTH: HBEC co-expressing TWIST1 wt and HRas G12V -EGFP; HFH: HBEC co-expressing TWIST1 F187G and HRas G12V -EGFP].

Article Snippet: Twist1 wt and Twist1 F191G exon 1 of founders were sequenced to validate the model by the Johns Hopkins Medicine Genetic Resources Core Facility, Baltimore, MD, USA ( ).

Techniques: In Vitro, Mutagenesis, Modification, Control, Transduction, Western Blot, Expressing, RNA Sequencing, Comparison, Plasmid Preparation

Journal: Neoplasia (New York, N.Y.)

Article Title: Twist1-induced suppression of oncogene-induced senescence in non-small cell lung cancer requires the transactivation domain of Twist1

doi: 10.1016/j.neo.2025.101179

Figure Lengend Snippet: The transactivation function of TWIST1 is required for TWIST1-induced suppression of OIS. Lung tumor tissue sections from CR, CRT and CRF191G mice (week 25 on doxycycline): ( A ) IHC (magnification 20x) for the expression of cell proliferation marker Ki67, cell death marker cleaved-caspase 3, and cell cycle arrest/senescence marker p16. ( B ) Quantifications of A. Data are presented as average values ± S.D. ( n = 3-4 animal subjects per time point; unpaired, two-tailed t- test: P > 0.05(ns)/ P < 0.05(*)/0.01(**)/0.001(***)). ( C-D ) At 72-hour post-adenoviral transduction for HRas G12V -EGFP or control-EGFP in HBEC— Control , HBEC- TWIST1 wt and HBEC- TWIST F187G : ( C ) Senescence-associated beta-galactosidase staining acquisition (magnification 10x) and quantification. Data are presented as average values ± S.D. ( n = 3; two-way anova test: P > 0.05(ns)/ P < 0.05(*)/0.01(**)/0.001(***)); ( D ) RT-PCR for cell cycle arrest/senescence markers p16. Data are presented as average values ± S.D. ( n = 4; unpaired, two-tailed t- test: P > 0.05(ns)/ P < 0.05(*)/0.01(**)/0.001(***)).

Article Snippet: Twist1 wt and Twist1 F191G exon 1 of founders were sequenced to validate the model by the Johns Hopkins Medicine Genetic Resources Core Facility, Baltimore, MD, USA ( ).

Techniques: Expressing, Marker, Two Tailed Test, Transduction, Control, Staining, Reverse Transcription Polymerase Chain Reaction

Journal: Neoplasia (New York, N.Y.)

Article Title: Twist1-induced suppression of oncogene-induced senescence in non-small cell lung cancer requires the transactivation domain of Twist1

doi: 10.1016/j.neo.2025.101179

Figure Lengend Snippet: The transactivation function of TWIST1 differentially regulates MYC transcriptional programs during HRas G12V oncogene-induced senescence. At 72-hours post-adenoviral transduction for HRas G12V -EGFP or control-EGFP in HBEC— Control , HBEC- TWIST1 wt and HBEC- TWIST F187G : ( A ) MYC Target gene panel using GeneGlobe Data Analysis Center (Qiagen) shows a downregulation of a MYC transcriptional targets with Control and TWIST F187G conditions that is reversed with TWIST1 wt as shown in volcano-plots ( n = 3; fold regulation threshold was 1.5 and p- value threshold was 0.05); ( B ) Clustergram of significantly upregulated and downregulated MYC target gene expression under HRas G12V (34 genes= 40.48 % of 84 genes tested), ( n = 3; fold regulation threshold was 1.5 and p- value threshold was 0.05); and ( C ) Venn-diagram of MYC targets significantly upregulated and downregulated genes (HBEC- TWIST1 wt -HRas G12V differentially modulated (highlighted numbers) 15.48 % of the tested genes or 38.24 % of the significantly modulated genes under HRas G12V ).

Article Snippet: Twist1 wt and Twist1 F191G exon 1 of founders were sequenced to validate the model by the Johns Hopkins Medicine Genetic Resources Core Facility, Baltimore, MD, USA ( ).

Techniques: Transduction, Control, Targeted Gene Expression

Journal: Neoplasia (New York, N.Y.)

Article Title: Twist1-induced suppression of oncogene-induced senescence in non-small cell lung cancer requires the transactivation domain of Twist1

doi: 10.1016/j.neo.2025.101179

Figure Lengend Snippet: The HBP/O-GlcNAc and MYC axes are required for TWIST1-mediated suppression of OIS. At 72-hours post-adenoviral transduction for HRas G12V -EGFP or control-EGFP in HBEC— Control , HBEC- TWIST1 wt and HBEC- TWIST F187G : Senescence-associated beta-galactosidase staining images and quantification of cells treated 48-hours with ( A-B ) OGA inhibitor TMG (50μ M), ( A-C ) GFPT2 inhibitor DON (5μ M), ( A-D ) OGT inhibitor OSMI1 (7.5μ M), and ( E-F ) MYC inhibitor MYCi975 (0.25μ M or 0.6μ M or 1.2μ M). Data are presented as average values ± S.D. ( n = 3; two-way anova test: P > 0.05(ns)/ P < 0.05(*)/0.01(**)/0.001(***)). [Overall, parental HBEC cells were globally following the same profile as the conditions HBEC— Control, ].

Article Snippet: Twist1 wt and Twist1 F191G exon 1 of founders were sequenced to validate the model by the Johns Hopkins Medicine Genetic Resources Core Facility, Baltimore, MD, USA ( ).

Techniques: Transduction, Control, Staining