Journal: Nature Communications

Article Title: Identification of recurrent FHL2-GLI2 oncogenic fusion in sclerosing stromal tumors of the ovary

doi: 10.1038/s41467-019-13806-x

Figure Lengend Snippet: a Cell titer blue proliferation assay of immortalized mesenchymal stem cells (MSCs), HEK-293, medulloblastoma (DAOY) and human basal cell carcinoma (BCC) cells stably expressing empty vector (control), wild-type FHL2 (FHL2), wild-type GLI2 (GLI2), truncated GLI2 (tGLI2) or FHL2-GLI2 treated with 250 nM Vismodegib or vehicle control (DMSO). b Representative images of colony formation assay of MSC, HEK-293, DAOY, and BCC cells stably expressing control, FHL2, GLI2, tGLI2, or FHL2-GLI2 treated with Vismodegib 500 nM or vehicle control (DMSO). Scale bars, 5 mm. Quantification of the number of colonies/well compared to control (bottom). c Wound healing assay of MSC, HEK-293, DAOY, and BCC cells stably expressing control, FHL2, GLI2, tGLI2, or FHL2-GLI2 treated with 250 nM Vismodegib or vehicle control (DMSO). The migratory effect/wound area was assessed at 0 and 24 h and quantified compared to DMSO (bottom). Vismo, Vismodegib. Scale bars, 500 μm. In a – c , data are representative of at least three independent experiments. Error bars, s.d. of mean; n.s., not significant; * P < 0.05, ** P < 0.01, *** P < 0.001; two-tailed unpaired t -test.

Article Snippet: The smoothened inhibitors Cyclopamine (Selleckchem, S1146), Vismodegib (Cellagen tech, C4044–5) and the GLI inhibitor GANT61 (Tocris, 3191) were resuspended in DMSO and used in in vitro proliferation, colony formation and scratch wound healing assays at 5–10 μM, 250–500 nM, and 5–10 μM, respectively.

Techniques: Proliferation Assay, Stable Transfection, Expressing, Plasmid Preparation, Control, Colony Assay, Wound Healing Assay, Two Tailed Test

Journal: Molecular Cancer

Article Title: Hedgehog signaling drives glial cell plasticity and oncogenic reprogramming in gastroenteropancreatic neuroendocrine neoplasms

doi: 10.1186/s12943-026-02611-y

Figure Lengend Snippet: Patient-derived PanNET tumoroids express HH signaling proteins and respond to HH pathway activation and inhibition. A Combined fluorescence and phase-contrast images of 21-day-old human PanNET tumoroids (PanNET3) stained for chromogranin A (CHGA) and HH proteins (PTCH1, SHH). B Four patient-derived PanNET tumoroid lines (PanNET1–4) were exposed to recombinant human SHH N-terminal peptide (100 ng/mL) or the SMO inhibitor vismodegib (VISMO, 20 µM) for 72 h and changes in mRNA expression were analyzed by RT-qPCR. mRNA changes were normalized to HPRT1 expression and DMSO vehicle control. ( n = 4 unique patient lines). * = p < 0.05, ** = p < 0.01, **** = p < 0.0001 by Two-way ANOVA with Sidak post-test. C EdU labeling showing proliferation of dissociated PanNET3 cells following 5-day exposure to: the HH agonists SHH-N (100 ng/mL) and SAG (10 nM); inhibitors of the canonical HH signaling pathway vismodegib (20 µM) and sonidegib (10 nM); or inhibitors of the GLI1/2 effectors GANT61 (10 µM) and itraconazole (ITZ, 1 µM). D Quantitation of the percentage of EdU-positive PanNET tumor cells following 5-day treatment. ( n = 3 replicates from one patient tumoroid line). * = p < 0.05, by One-way ANOVA with Tukey post-test. E SHH or SAG were co-administered with the respective pharmacologic inhibitors and EdU uptake was evaluated after 7 days. F Immunofluorescent images of CHGA, SHH, and PTCH1 expression in a second PanNET tumoroid line (PanNET5). G , H EdU labeling was evaluated in PanNET5 tumoroids following 7-day treatment with SHH-N (200 ng/mL) and SAG (20 nM); inhibitors of the canonical HH signaling pathway vismodegib (20 µM) and sonidegib (10 nM); or inhibitors of the GLI1/2 effectors GANT61 (10 µM) and itraconazole (ITZ, 5 µM). I Crystal violet staining of human BON-1 PanNET cells after 48 h treatment. J BrdU incorporation in BON-1 cells after 48 h treatment with HH agonists SHH-N and SAG, ( K ) SMO inhibitors vismodegib and sonidegib, and ( L ) inhibitors of GLI1/2 signaling. M Immunofluorescent images of CHGA, SHH, and PTCH1 expression in tumoroids derived from a metastatic ileal NET (IL-NET-met1). N , O EdU labeling was assayed in the IL-NET-met1 tumoroid line following 7-day treatment with the same drug concentrations used for PanNET5. * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001 by One-way ANOVA with Dunnett post-test

Article Snippet: Drug compounds used in the studies include: human recombinant SHH N-terminal peptide (R&D Systems, Cat# 1845-GMP), SAG (Tocris, Cat# 4366), vismodegib (Tocris, Cat# 7710), sonidegib (Tocris, Cat# 7826), GANT61 (Tocris, Cat# 3191), and itraconazole (Tocris, Cat# 5981).

Techniques: Derivative Assay, Activation Assay, Inhibition, Fluorescence, Staining, Recombinant, Expressing, Quantitative RT-PCR, Control, Labeling, Quantitation Assay, BrdU Incorporation Assay

Journal: Molecular Cancer

Article Title: Hedgehog signaling drives glial cell plasticity and oncogenic reprogramming in gastroenteropancreatic neuroendocrine neoplasms

doi: 10.1186/s12943-026-02611-y

Figure Lengend Snippet: HH signaling regulates the growth of GFAP ΔMen1 and Sox10 ΔMen1 pancreatic NET tumoroids. A Phase contrast and ( B ) fluorescence images of PanNET tumoroids from Sox10-Cre; Men1 FL/FL ; LSL-tdTomato mice. Tumoroids were imaged after 72 h exposure to: the HH agonists SHH-N (100 ng/mL) and SAG (10 nM); inhibitors of the canonical HH signaling pathway vismodegib (20 µM) and sonidegib (10 nM); or inhibitors of the GLI1/2 effectors GANT61 (10 µM) and itraconazole (ITZ, 1 µM). C TdTomato fluorescence intensity was used to measure PanNET tumoroid growth in the presence of HH pathway agonists or ( D ) inhibitors of the canonical and ( E ) non-canonical HH signaling pathways. Fluorescence signal is compared to DMSO vehicle control. ( n = 3 replicates in two unique mouse PanNET tumoroid lines). ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001 by One-way ANOVA with Dunnett post-test. F BrdU incorporation was used to measure tumoroid proliferation in GFAP ΔMen1 and Sox10 ΔMen1 PanNET tumoroids after 72 h treatment. ( n = 4). ** = p < 0.01, **** = p < 0.0001 by One-way ANOVA with Dunnett post-test. G Relative fold-change in Chga mRNA levels in PanNET tumoroids following 72 h treatment. ( n = 5). H Western blot analysis of HH pathway proteins in PanNET tumoroids after 72 h treatment. I Quantitation of SHH protein expression normalized to beta-actin and DMSO vehicle control from the western blot analysis in panel ( H ). ( n = 3). J Western blot analysis and ( K ) associated quantitation of phosphorylated and total ERK and AKT growth pathways in PanNET tumoroids after 72 h treatment. ( n = 3). * = p < 0.05, ** = p < 0.01, *** = p < 0.001 by Kruskal-Wallis test

Article Snippet: Drug compounds used in the studies include: human recombinant SHH N-terminal peptide (R&D Systems, Cat# 1845-GMP), SAG (Tocris, Cat# 4366), vismodegib (Tocris, Cat# 7710), sonidegib (Tocris, Cat# 7826), GANT61 (Tocris, Cat# 3191), and itraconazole (Tocris, Cat# 5981).

Techniques: Fluorescence, Protein-Protein interactions, Control, BrdU Incorporation Assay, Western Blot, Quantitation Assay, Expressing

Journal: Molecular Cancer

Article Title: Hedgehog signaling drives glial cell plasticity and oncogenic reprogramming in gastroenteropancreatic neuroendocrine neoplasms

doi: 10.1186/s12943-026-02611-y

Figure Lengend Snippet: GFAP ΔMen1 DNETs and jejunal NETs are sensitive to HH pathway activation and inhibition. A Phase contrast and ( B ) fluorescence images of DNET tumoroids from a GFAP-Cre; Men1 FL/FL ; LSL-tdTomato mouse. Tumoroids were imaged after 72 h exposure to: the HH agonists SHH-N (100 ng/mL) and SAG (10 nM); inhibitors of the canonical HH signaling pathway vismodegib (20 µM) and sonidegib (10 nM); or inhibitors of the GLI1/2 effectors GANT61 (10 µM) and itraconazole (ITZ, 1 µM). C TdTomato fluorescence intensity was used to measure DNET tumoroid growth in the presence of HH pathway inhibitors. Fluorescence signal is compared to DMSO vehicle control. ( n = 3 replicates using one mouse DNET tumoroid line). ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001 by One-way ANOVA with Dunnett post-test. D Relative BrdU incorporation in a second GFAP ΔMen1 DNET tumoroid line and ( E ) a jejunal tumoroid line (J-NET) after 72 h treatment. ( n = 4). ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001 by One-way ANOVA with Dunnett post-test. F Western blot analysis of SHH, ERK, and AKT growth pathways in J-NET tumoroids after 72 h treatment. G Western blot quantitation of SHH and ( H ) phosphorylated and total ERK and AKT proteins normalized to GAPDH and DMSO vehicle control. ( n = 3). * = p < 0.05 by Kruskal-Wallis test. I Crystal violet staining of mouse STC-1 SI-NET cells after 48 h treatment with agonists and inhibitors of the HH signaling pathway. J BrdU incorporation in STC-1 cells after 48 h treatment with HH agonists SHH-N and SAG, ( K ) SMO inhibitors vismodegib and sonidegib, and ( L ) inhibitors of GLI1/2 signaling. * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001 by One-way ANOVA with Dunnett post-test

Article Snippet: Drug compounds used in the studies include: human recombinant SHH N-terminal peptide (R&D Systems, Cat# 1845-GMP), SAG (Tocris, Cat# 4366), vismodegib (Tocris, Cat# 7710), sonidegib (Tocris, Cat# 7826), GANT61 (Tocris, Cat# 3191), and itraconazole (Tocris, Cat# 5981).

Techniques: Activation Assay, Inhibition, Fluorescence, Control, BrdU Incorporation Assay, Western Blot, Quantitation Assay, Staining

Journal: PLoS ONE

Article Title: Definition of Critical Periods for Hedgehog Pathway Antagonist-Induced Holoprosencephaly, Cleft Lip, and Cleft Palate

doi: 10.1371/journal.pone.0120517

Figure Lengend Snippet: Single doses of vismodegib were administered at discrete time points indicated by tick marks on the x-axis, including: GD7.0, 7.25, 7.5, 7.75, 8.0, 8.25, 8.5, 8.625, 8.75, 8.875, 9.0, 9.25, 9.5, 9.75, and 10.0. Cyclopamine was administered by subcutaneous infusion from GD8.25 to ~9.375. Representative examples of distinct face and palate phenotypes are shown, including apparently normal (Normal), HPE, CL/P, and CPO. Note that lateral lip clefts resulting from acute vismodegib exposure typically extended into the primary palate (D’), while those resulting from cyclopamine exposure extended into both the primary and secondary palate (F’). The penetrance of HPE, CL/P, and CPO phenotypes resulting from stage-specific vismodegib exposure is shown in the graph. 5–7 litters were examined for each exposure permutation.

Article Snippet: Vismodegib (Toronto Research Chemicals) was suspended as 3mg/ml in 0.5% methyl cellulose (Sigma) with 0.2% Tween (Sigma).

Techniques:

Journal: PLoS ONE

Article Title: Definition of Critical Periods for Hedgehog Pathway Antagonist-Induced Holoprosencephaly, Cleft Lip, and Cleft Palate

doi: 10.1371/journal.pone.0120517

Figure Lengend Snippet: Superior views of dissected brains are shown for a vehicle-exposed normal animal (A) and for representative examples of animals with vismodegib-induced HPE (B), cyclopamine-induced CL/P (C), and vismodegib-induced CPO (D). Severe hypoplasia of the cerebral cortices (cc) and olfactory bulb (ofb) absence is apparent in the animal with HPE. In animals with CL/P and CPO, the cerebral cortices appear to be of approximately normal size but the olfactory bulbs are hypoplastic. Serial coronal sections of comparably classified animals are shown in E-P. Notable HPE-associated features include a single central nasal passage with nasal septum (ns) cartilage absence (black arrow), olfactory bulb agenesis (J), and a single telencephalic vesicle (N). Grossly normal division of the olfactory bulbs (K, L) and cerebral cortices (O, P), and apparent forebrain septal (s) region hyperplasia (white arrows) is observed in animals with CL/P and CPO. (t) Tongue, (e) eye, (sp) secondary palate.

Article Snippet: Vismodegib (Toronto Research Chemicals) was suspended as 3mg/ml in 0.5% methyl cellulose (Sigma) with 0.2% Tween (Sigma).

Techniques:

Journal: PLoS ONE

Article Title: Definition of Critical Periods for Hedgehog Pathway Antagonist-Induced Holoprosencephaly, Cleft Lip, and Cleft Palate

doi: 10.1371/journal.pone.0120517

Figure Lengend Snippet: In vehicle-exposed embryos at GD14.5 (A) the secondary palatal shelves have approximated and made contact at the midline. In affected cyclopamine-exposed embryos with cleft lip (B), palatal shelves are widely spaced and deficient in width. In vismodegib-exposed embryos (C), secondary palatal shelves have also elevated but are deficient in both length and width. Length (D) and width (E) measurements (arbitrary units), as depicted by the dashed calipers, were made on light microscopy images . Shelf width was determined at 1/3 shelf length from the most rostral aspect. *** p<0.001, **** p<0.0001

Article Snippet: Vismodegib (Toronto Research Chemicals) was suspended as 3mg/ml in 0.5% methyl cellulose (Sigma) with 0.2% Tween (Sigma).

Techniques: Light Microscopy

Journal: PLoS ONE

Article Title: Definition of Critical Periods for Hedgehog Pathway Antagonist-Induced Holoprosencephaly, Cleft Lip, and Cleft Palate

doi: 10.1371/journal.pone.0120517

Figure Lengend Snippet: Along with a vehicle-exposed control (A), representative examples of phenotypic outcomes are shown with numbers indicating the gestational stage of acute vismodegib administration. Later exposure was associated with forelimb ectrodactyly, as exhibited bilaterally in fetuses exposed from 9.25 to 9.75 (arrows point to absent fifth digits on the right limb). Kinked tail phenotypes were caused by exposure between GD9.5 and 10.0 (arrowheads). Edema is also apparent in fetuses exposed at GD9.75 and 10.0. For each treatment group the number of litters and fetuses examined, mean litter size and crown-rump length, and the incidence of edema, forelimb ectrodactyly, and kinked tail defects are presented in .

Article Snippet: Vismodegib (Toronto Research Chemicals) was suspended as 3mg/ml in 0.5% methyl cellulose (Sigma) with 0.2% Tween (Sigma).

Techniques: Control

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: CD44 expression denotes a subpopulation of gastric cancer cells in which Hedgehog signaling promotes chemotherapy resistance

doi: 10.1158/1078-0432.CCR-14-0011

Figure Lengend Snippet: Hedgehog pathway inhibition with Smo shRNA or vismodegib blocks spheroid formation. A. Western blot demonstrating knockdown of Smo in gastric cancer cell lines AGS, MKN-45, and N87 following transduction with Smo shRNA (Smo.shRNA) lentivirus compared to scrambled shRNA control (Scr.shRNA) lentivirus. B. Immunofluorescence photos for CD44 (green) and nuclei (blue) of gastric cancer cell lines treated with Smo.shRNA or Scr.shRNA and grown in spheroid formation conditions or treated with vismodegib (10 μM) or carrier (DMSO) and grown in spheroid formation conditions. C. Single cell assay of spheroid cells showing diameter of spheroids at selected time points following treatment with Smo.shRNA vs. Scr.shRNA or vismodegib (Vis, 10 μM) vs. carrier (DMSO). Bars represent standard deviation.

Article Snippet: Vismodegib was purchased from LC Laboratories (Woburn, MA).

Techniques: Inhibition, shRNA, Western Blot, Transduction, Immunofluorescence, Standard Deviation

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: CD44 expression denotes a subpopulation of gastric cancer cells in which Hedgehog signaling promotes chemotherapy resistance

doi: 10.1158/1078-0432.CCR-14-0011

Figure Lengend Snippet: CD44(+) gastric cancer cells demonstrate chemotherapy resistance, which can be reversed with Hedgehog pathway inhibition. A. Immunofluorescence images of CD44 (green) and nuclei (blue) for CD44(+) and CD44(−) cells following magnetic bead sorting of AGS, MKN-45, and N87 gastric cancer cell lines. B. Western blot showing expression of Hedgehog pathway proteins Sonic hedgehog (Shh), Patch 1 (Ptch1), Smoothened (Smo), and Gli1 in CD44(+) and CD44(−) cells. Proliferation assay for CD44(+) and CD44(−) cells (C) and spheroid cells and monolayer cells (D) following treatment with 5-fluorouracil (5-FU) or cisplatin chemotherapy and vismodegib (Vis, 10 μM) compared to DMSO. Bars represent standard deviation.

Article Snippet: Vismodegib was purchased from LC Laboratories (Woburn, MA).

Techniques: Inhibition, Immunofluorescence, Western Blot, Expressing, Proliferation Assay, Standard Deviation

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: CD44 expression denotes a subpopulation of gastric cancer cells in which Hedgehog signaling promotes chemotherapy resistance

doi: 10.1158/1078-0432.CCR-14-0011

Figure Lengend Snippet: Gastric cancer spheroid cells demonstrate increased migration, colony formation, and anchorage independent growth, which can be attenuated by Hedgehog pathway inhibition. A. Photos and graphs of migration assay of gastric cancer monolayer cells and spheroid cells treated with Smo shRNA (Smo.shRNA) compared to control (Scr.shRNA) or vismodegib (10 μM) compared to DMSO. B. Colony formation assay of gastric cancer monolayer cells and spheroid cells treated with Smo shRNA (Smo.shRNA) compared to control (Scr.shRNA) or vismodegib (10 μM) compared to DMSO. C. Photos and graph of soft agar assay of gastric cancer monolayer cells and spheroid cells treated with Smo shRNA (Smo.shRNA) compared to control (Scr.shRNA). Bars represent standard deviation.

Article Snippet: Vismodegib was purchased from LC Laboratories (Woburn, MA).

Techniques: Migration, Inhibition, shRNA, Colony Assay, Soft Agar Assay, Standard Deviation

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: CD44 expression denotes a subpopulation of gastric cancer cells in which Hedgehog signaling promotes chemotherapy resistance

doi: 10.1158/1078-0432.CCR-14-0011

Figure Lengend Snippet: CD44 may be a response biomarker in advanced gastric and gastroesophageal cancer patients treated with chemotherapy with or without vismodegib. A. Schema of the clinical trial. B. CD44 immunohistochemistry of patient tumor samples showing low, intermediate, and high CD44 expression. C. Kaplan-Meier overall survival curves for patients receiving chemotherapy alone and patients receiving chemotherapy plus vismodegib stratified by low and high CD44 score.

Article Snippet: Vismodegib was purchased from LC Laboratories (Woburn, MA).

Techniques: Biomarker Assay, Immunohistochemistry, Expressing

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: CD44 expression denotes a subpopulation of gastric cancer cells in which Hedgehog signaling promotes chemotherapy resistance

doi: 10.1158/1078-0432.CCR-14-0011

Figure Lengend Snippet: CD44 score, progression, and survival

Article Snippet: Vismodegib was purchased from LC Laboratories (Woburn, MA).

Techniques:

Journal: JAAD Case Reports

Article Title: Characterization of intractable diarrhea resulting from vismodegib treatment for basal cell nevus syndrome

doi: 10.1016/j.jdcr.2022.07.030

Figure Lengend Snippet: Colonoscopy findings of vismodegib-induced diarrhea showing ( A ) ilium with mucosal edema and granularity, ( B ) transverse colon with mildly granular mucosa and loss of vascular pattern, and ( C ) descending colon showing mildly granular mucosa with loss of vascular pattern.

Article Snippet: Vismodegib (Erivedge, Genentech) is a Food and Drug Administration–approved systemic hedgehog inhibitor used in the treatment of advanced BCCs, including the multiple BCCs found in Gorlin syndrome.

Techniques:

Journal: JAAD Case Reports

Article Title: Characterization of intractable diarrhea resulting from vismodegib treatment for basal cell nevus syndrome

doi: 10.1016/j.jdcr.2022.07.030

Figure Lengend Snippet: Microscopic findings of vismodegib-induced diarrhea in the ileum. Ileum biopsy sepcimens showing ( A ) mild villous blunting, ( B ) focal epithelial apoptosis ( yellow arrows ) and active ileitis ( red arrow ), and ( C ) focal active ileitis. A to C, Hematoxylin-eosin stain; original magnifications: A, ×10; B, ×40; C, ×40.

Article Snippet: Vismodegib (Erivedge, Genentech) is a Food and Drug Administration–approved systemic hedgehog inhibitor used in the treatment of advanced BCCs, including the multiple BCCs found in Gorlin syndrome.

Techniques: Staining

Journal: JAAD Case Reports

Article Title: Characterization of intractable diarrhea resulting from vismodegib treatment for basal cell nevus syndrome

doi: 10.1016/j.jdcr.2022.07.030

Figure Lengend Snippet: Microscopic findings of vismodegib-induced diarrhea in the colon. Colon biopsy sepcimen showing ( A ) the right colon with mild crypt branching and irregularity; ( B ) the right colon with epithelial apoptosis and mildly increased eosinophils ( the yellow arrow highlights a focus of epithelial apoptosis); ( C ) the left colon with active inflammation with neutrophils and eosinophils within an atrophic crypt, ( D ) the left colon with Paneth cell metaplasia, indicative of chronic damage; ( E ) the rectum demonstrating architectural distortion with crypt branching and irregular crypts; ( F ) the rectum demonstrating that epithelial apoptosis was present ( yellow arrows highlights foci of epithelial apoptosis). A to F, Hematoxylin-eosin stain; original magnifications: A, ×10; B, ×40; C, ×40; D, ×40; E, ×10; F, ×40.

Article Snippet: Vismodegib (Erivedge, Genentech) is a Food and Drug Administration–approved systemic hedgehog inhibitor used in the treatment of advanced BCCs, including the multiple BCCs found in Gorlin syndrome.

Techniques: Staining

Journal: American Journal of Cancer Research

Article Title: Anticancer therapeutics: a brief account on wide refinements

doi:

Figure Lengend Snippet: List of anti-angiogenic drugs approved by the FDA for the treatment of cancer

Article Snippet: , Vismodegib (Eviredge, Genentech) , , .

Techniques: Bioprocessing, Recombinant