Journal: Acta Neuropathologica Communications

Article Title: Genome-wide CRISPR-Cas9 knockout screens identify DNMT1 as a druggable dependency in sonic hedgehog medulloblastoma

doi: 10.1186/s40478-024-01831-x

Figure Lengend Snippet: DNMT1 and SMO inhibition synergize to block SHH-MB growth in vivo. a Schematic overview illustrating tumor induction and treatment timeline in Math1-creER T2 ::SmoM2 Fl/ + mice. b Kaplan–Meier curves of Math1-creER T2 ::SmoM2 Fl/ + mice treated with 5-AzaC monotherapy (n = 19), LDE225 monotherapy (n = 19), drug combination treatment (n = 19), and vehicle control-treated mice (n = 18). Vertical dashed line represents the last day of treatment. Significance in survival as compared to vehicle-treated mice was determined using the log rank (Mantel-Cox) test. c Representative H&E stainings of cerebellar tumors from Math1-creER T2 ::SmoM2 Fl/ + mice from indicated treatment groups on the last day of treatment (P68), as well as immunohistochemistry for Ki67 in these tumors. d Quantification of Ki67 shown in c (n = 3, Fisher´s exact test). e Western blot analysis of GLI1 and PCNA proteins deriving from harvested tumor tissue from Math1-creER T2 ::SmoM2 Fl/ + mice at P68. 4 × magnification, scale bar, 500µm; 20 × magnification, scale bar, 50µm. Graph displays mean ± SD. * p ≤ 0.05, ** p ≤ 0.01, **** p ≤ 0.0001

Article Snippet: The compounds tested, sonidegib, vismodegib and 5-azacytidine, were purchased from Selleckchem and diluted according to manufacturer ́s instructions.

Techniques: Inhibition, Blocking Assay, In Vivo, Control, Immunohistochemistry, Western Blot

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: PLOS ONE

Article Title: Analysis of residual disease in periocular basal cell carcinoma following hedgehog pathway inhibition: Follow up to the VISORB trial

doi: 10.1371/journal.pone.0265212

Figure Lengend Snippet: A) Pathology results for excision samples after vismodegib treatment (error bars indicate 95% CI, n = 9 (left column), n = 18 (right column)), B) Pre- and post-vismodegib histology, C) Average total area and D) Average lesion size (pixels) of keratin 5 positive staining (normalized to section length, error bars indicate SEM, unpaired t-test *p<0.05, n = 7 (left column), n = 11 (right column)), E) Clinical photo, histology, and keratin staining pre and post-vismodegib treatment of patients scored as “no sign of disease” (black arrow–tumor location, yellow box—area of peripheral palisading).

Article Snippet: Vismodegib (Genentech Inc.) is a small-molecule SMO inhibitor approved by the United States Food and Drug Administration (FDA) to treat advanced and metastatic BCC [ ].

Techniques: Staining

Journal: PLOS ONE

Article Title: Analysis of residual disease in periocular basal cell carcinoma following hedgehog pathway inhibition: Follow up to the VISORB trial

doi: 10.1371/journal.pone.0265212

Figure Lengend Snippet: Keratin 5 expression in post-vismodegib excision samples.

Article Snippet: Vismodegib (Genentech Inc.) is a small-molecule SMO inhibitor approved by the United States Food and Drug Administration (FDA) to treat advanced and metastatic BCC [ ].

Techniques: Expressing

Journal: PLOS ONE

Article Title: Analysis of residual disease in periocular basal cell carcinoma following hedgehog pathway inhibition: Follow up to the VISORB trial

doi: 10.1371/journal.pone.0265212

Figure Lengend Snippet: A) Gli1 expression (dots/nuclei) pre- and post-vismodegib treatment in patient samples scored as disease present (red) or no sign of disease (green) (error bars indicate SEM, unpaired t-test *p<0.05, n = 7,6,9,11 columns left to right), B) Gli1 expression (brown dots) in excision samples from “disease present” (left panels) and “no sign of disease” (right panels) samples. C) Gli1 expression (left panels, brown) and proliferation (Ki67, green) in keratin 5 positive (red) persistent lesions from samples scored as disease present (top panels) and no sign of disease (bottom panels).

Article Snippet: Vismodegib (Genentech Inc.) is a small-molecule SMO inhibitor approved by the United States Food and Drug Administration (FDA) to treat advanced and metastatic BCC [ ].

Techniques: Expressing

Journal: PLOS ONE

Article Title: Analysis of residual disease in periocular basal cell carcinoma following hedgehog pathway inhibition: Follow up to the VISORB trial

doi: 10.1371/journal.pone.0265212

Figure Lengend Snippet: Gli1 expression in post-vismodegib excision samples.

Article Snippet: Vismodegib (Genentech Inc.) is a small-molecule SMO inhibitor approved by the United States Food and Drug Administration (FDA) to treat advanced and metastatic BCC [ ].

Techniques: Expressing

Journal: PLOS ONE

Article Title: Analysis of residual disease in periocular basal cell carcinoma following hedgehog pathway inhibition: Follow up to the VISORB trial

doi: 10.1371/journal.pone.0265212

Figure Lengend Snippet: A) Clinical photos and Gli1 expression (brown dots) in a patient pre- (left panels), post-vismodegib (center panels), and after recurrence (right panels), B) Allele frequency of PTCH1 and SMO variants pre-, post-vismodegib, and after recurrence (*variant not detected)(yellow star–predicted pathogenic variant, red star–known pathogenic variant).

Article Snippet: Vismodegib (Genentech Inc.) is a small-molecule SMO inhibitor approved by the United States Food and Drug Administration (FDA) to treat advanced and metastatic BCC [ ].

Techniques: Expressing, Variant Assay