Journal: Nature Communications

Article Title: Mapping the landscape of genetic dependencies in chordoma

doi: 10.1038/s41467-023-37593-8

Figure Lengend Snippet: a Colony formation assays of chordoma and non-chordoma (negative control A2058; positive control MDA-MB-468) cell lines treated with indicated concentrations of RMC-4550 for 14 days. b Viability of chordoma and non-chordoma (negative control A2058; positive control MDA-MB-468) cell lines treated with indicated concentrations of SHP2 inhibitors RMC-4550 and SHP099 and assayed for cell viability after 6 days with CellTiter-Glo. Response data were represented by a fitted curve to the mean fractional viability at each concentration relative to vehicle-treated cells; error bars represent the s.e.m. ( n = 4 biological samples measured in parallel). c Immunoblot analysis of chordoma and non-chordoma (negative control A2058; positive control MDA-MB-468) cell lines treated with indicated concentrations of RMC-4550, SHP099, or DMSO for 2 h. d Tumor proliferation in mice engrafted with chordoma cells (U-CH1 cell line-derived xenograft, CF539 PDX, or CF466 PDX) and treated with a SHP2 inhibitor (RMC-4550 or TNO155). Points represent the mean tumor volume ± s.e.m. ( n = 4 (control) or 5 (compound) tumors for each arm of the U-CH1/RMC-4550 study; n = 6 (compound) or 7 (control) tumors for each arm of the U-CH1/TNO155 study; n = 6 (control) or 7 (compound) tumors for each arm of the CF539 study; n = 7 tumors for each arm of the CF466 study). n.s., not significant, * P < 0.05, **** P < 0.0001, derived from a two-way analysis of variance (ANOVA) with repeated measures. P values for the time × treatment interaction (relative to the control condition) are indicated. Additional details of P values and effect sizes are reported in Supplementary Data . Source data are provided as a Source Data file. See also related Supplementary Data .

Article Snippet: TNO155 used for ex vivo studies was purchased from Selleck Chemicals, and that used for in vivo studies was purchased from MedChemExpress.

Techniques: Negative Control, Positive Control, Concentration Assay, Western Blot, Derivative Assay, Control

Journal: Nature Communications

Article Title: Mapping the landscape of genetic dependencies in chordoma

doi: 10.1038/s41467-023-37593-8

Figure Lengend Snippet: a Colony formation assays of chordoma and non-chordoma (negative control A2058; positive control MDA-MB-468) cell lines treated with indicated concentrations of RMC-4550 for 14 days. b Viability of chordoma and non-chordoma (negative control A2058; positive control MDA-MB-468) cell lines treated with indicated concentrations of SHP2 inhibitors RMC-4550 and SHP099 and assayed for cell viability after 6 days with CellTiter-Glo. Response data were represented by a fitted curve to the mean fractional viability at each concentration relative to vehicle-treated cells; error bars represent the s.e.m. ( n = 4 biological samples measured in parallel). c Immunoblot analysis of chordoma and non-chordoma (negative control A2058; positive control MDA-MB-468) cell lines treated with indicated concentrations of RMC-4550, SHP099, or DMSO for 2 h. d Tumor proliferation in mice engrafted with chordoma cells (U-CH1 cell line-derived xenograft, CF539 PDX, or CF466 PDX) and treated with a SHP2 inhibitor (RMC-4550 or TNO155). Points represent the mean tumor volume ± s.e.m. ( n = 4 (control) or 5 (compound) tumors for each arm of the U-CH1/RMC-4550 study; n = 6 (compound) or 7 (control) tumors for each arm of the U-CH1/TNO155 study; n = 6 (control) or 7 (compound) tumors for each arm of the CF539 study; n = 7 tumors for each arm of the CF466 study). n.s., not significant, * P < 0.05, **** P < 0.0001, derived from a two-way analysis of variance (ANOVA) with repeated measures. P values for the time × treatment interaction (relative to the control condition) are indicated. Additional details of P values and effect sizes are reported in Supplementary Data . Source data are provided as a Source Data file. See also related Supplementary Data .

Article Snippet: TNO155 used for ex vivo studies was purchased from Selleck Chemicals, and that used for in vivo studies was purchased from MedChemExpress.

Techniques: Negative Control, Positive Control, Concentration Assay, Western Blot, Derivative Assay, Control

Journal: Molecular Cancer

Article Title: Targeting KRAS mutant cancers: from druggable therapy to drug resistance

doi: 10.1186/s12943-022-01629-2

Figure Lengend Snippet: Summary of KRAS mutation cancers therapeutics

Article Snippet: The SHP2 inhibitor developed by Novartis, TNO155, inhibits MAPK signaling and enhances the efficacy of KRAS (G12C) inhibitors against KRAS (G12C) lung and colorectal cancers.

Techniques: Mutagenesis, In Vitro, In Vivo, Animal Model, Inhibition, Small Interfering RNA

Journal: Molecular Cancer

Article Title: Targeting KRAS mutant cancers: from druggable therapy to drug resistance

doi: 10.1186/s12943-022-01629-2

Figure Lengend Snippet: Clinical trials targeting KRAS mutation cancers

Article Snippet: The SHP2 inhibitor developed by Novartis, TNO155, inhibits MAPK signaling and enhances the efficacy of KRAS (G12C) inhibitors against KRAS (G12C) lung and colorectal cancers.

Techniques: Clinical Proteomics, Mutagenesis

Journal: The Journal of Clinical Investigation

Article Title: Off-target autophagy inhibition by SHP2 allosteric inhibitors contributes to their antitumor activity in RAS-driven cancers

doi: 10.1172/JCI177142

Figure Lengend Snippet: ( A ) RFP-LAMP1–expressing U2OS cells were treated with DMSO, 10 μM SHP099, or IACS-13909 for 3 hours. Representative images of LAMP1 (red, lysosome marker), SHP099 (green and blue), or IACS-13909 (blue) and merged channels are displayed. Scale bars: 10 μm. ( B ) RFP-LAMP1–expressing WT or SHP2-KO HEK293 cells were treated with 10 μM SHP099 or IACS-13909 for 3 hours. Representative images of LAMP1 (red, lysosome marker), SHP099 (green and blue), or IACS-13909 (blue) and merged channels are displayed. Scale bars: 10 μm. ( C ) WT or SHP2 –/– HEK293 cells were treated with a series of SHP099 or IACS-13909 concentrations for 6 hours. Total lysates were used for immunoblots. ( D ) A375 cells were treated with a series of SHP099 concentrations with or without the presence of 20 μM CQ for 6 hours. Total lysates were used for immunoblots. ( E ) U2OS cells were treated with 10 μM SHP099, TNO155, or IACS-13909 for 6 hours and visualized with transmission electronic microscopy. Scale bars: 1 μm. ( F ) HEK293 cells were treated with 10 μM SHP099, RMC-4550, or IACS-13909 for 6 hours with or without 20 μM CQ pretreatment for 3 hours. Total lysates were used for immunoblots. Lanes were run on the same gel but were noncontiguous. ( G ) EGFP-mCherry-LC3–expressing U2OS cells were treated with 0.5 or 5 μM IACS-13909 or DMSO for 6 hours. Representative images of EGFP (green, pH sensitive), mCherry (red, pH insensitive), and merged channels are displayed. Scale bars: 10 μm. Autophagic index indicates the ratio of the areas of mCherry + puncta to EGFP + puncta. Mean autophagic index is plotted, with each individual data point representing 1 analyzed cell field (5–10 fields total) from 3 independent experiments (labeled with different colors). Significance was determined by 1-way ANOVA followed by Dunnett’s multiple-comparison test. *** P < 0.001. ( H ) Autophagy inhibition levels were determined by quantification of LC3-II/I ratio changes in . To determine the EC 50 , a 5-fold increase of LC3-II/I ratio in comparison with DMSO was defined as 100% autophagy inhibition. Representative data from 3 independent experiments displayed for all panels.

Article Snippet: SHP099 (CT-SHP099), IACS-13909 (CT-IACS-13909), RMC-4550 (CT-RMC4550), JAB-3068 (CT-JAB3068), and TNO155 (CT-TNO155) were purchased from Chemietek.

Techniques: Expressing, Marker, Western Blot, Transmission Assay, Microscopy, Labeling, Comparison, Inhibition

Journal: The Journal of Clinical Investigation

Article Title: Off-target autophagy inhibition by SHP2 allosteric inhibitors contributes to their antitumor activity in RAS-driven cancers

doi: 10.1172/JCI177142

Figure Lengend Snippet: ( A ) Results of colony formation assay using WT or SHP2 –/– HEK293 cells with or without overexpression of the dominant-negative ATG4B C74A mutation and treated with DMSO or 1.25 or 2.5 μM IACS-13909 for 10 days. ( B ) Results of colony formation assay using a panel of cancer cell lines treated with DMSO or indicated compounds for 10 days. ( C ) Changes in tumor volume of MIA PaCa-2 xenografts on day 21 compared with day 1 after treatment with vehicle control, 50 mg/kg TNO155, 50 mg/kg IACS-13909, 50 mg/kg CQ, and CQ with TNO155 ( n = 6–10 per treatment group). Data are represented as means ± SD. Significance was determined by Brown-Forsythe and Welch’s ANOVA test followed by 2-stage linear step-up procedure of Benjamini, Krieger, and Yekutieli. * P < 0.05; ** P < 0.01; *** P < 0.001. Representative data from 3 independent experiments displayed for A and B .

Article Snippet: SHP099 (CT-SHP099), IACS-13909 (CT-IACS-13909), RMC-4550 (CT-RMC4550), JAB-3068 (CT-JAB3068), and TNO155 (CT-TNO155) were purchased from Chemietek.

Techniques: Colony Assay, Over Expression, Dominant Negative Mutation, Mutagenesis, Control

Journal: The Journal of Clinical Investigation

Article Title: Off-target autophagy inhibition by SHP2 allosteric inhibitors contributes to their antitumor activity in RAS-driven cancers

doi: 10.1172/JCI177142

Figure Lengend Snippet: ( A ) Results of colony formation assay using a panel of cancer cell lines treated with DMSO or indicated compounds for 10 days. ( B and C ) MIA PaCa-2 ( B ) or HCT 116 ( C ) cells were treated with DMSO, 10 nM trametinib, 1 μM TNO155, or IACS-13909 or combinations for the times indicated. Total lysates were used for immunoblots. ( D and E ) Changes in tumor volume of MIA PaCa-2 ( D ) and HCT 116 ( E ) xenografts on day 21 compared with day 1 after treatment with vehicle control, 40 mg/kg TNO155, 40 mg/kg IACS-13909, 0.25 mg/kg trametinib, or combinations ( n = 5–10). Total lysates from individual tumors were used for immunoblots. Data are represented as means ± SD. Significance was determined by Brown-Forsythe and Welch’s ANOVA test followed by 2-stage linear step-up procedure of Benjamini, Krieger, and Yekutieli. * P < 0.05; ** P < 0.01; *** P < 0.001. Representative data from 3 independent experiments displayed for A – C .

Article Snippet: SHP099 (CT-SHP099), IACS-13909 (CT-IACS-13909), RMC-4550 (CT-RMC4550), JAB-3068 (CT-JAB3068), and TNO155 (CT-TNO155) were purchased from Chemietek.

Techniques: Colony Assay, Western Blot, Control

Journal: Theranostics

Article Title: Conquering oncogenic KRAS and its bypass mechanisms

doi: 10.7150/thno.71260

Figure Lengend Snippet: List of therapeutic approaches targeting KRAS-RAF-MEK in clinical trials and FDA-approved

Article Snippet: MRTX849 (adagrasib) , KRAS G12C , KRAS G12C mutant advanced cancers , Ph1/2; with TNO155 (SHP2 inhibitor) , NCT04330664 , , Mirati.

Techniques: Clinical Proteomics, Mutagenesis

Journal: iScience

Article Title: Overcoming MET-mediated resistance in oncogene-driven NSCLC

doi: 10.1016/j.isci.2023.107006

Figure Lengend Snippet: The combination of targeted therapies with tepotinib or TNO155 decreases MAPK/PI3K downstream signaling more potently than either of the agents alone (A) HCC827 tetON-MET and NCI-H358 tetON-MET, as well as (B) NCI-H1781 tetON-MET and KM12 tetON-MET cells were induced with doxycycline for 48 h before they were treated with the respective inhibitors and combinations for 6 h. Western blot analysis was performed with the indicated antibodies.

Article Snippet: TNO155 , Synthesized at the healthcare business of Merck KGaA, Darmstadt, Germany , NA.

Techniques: Western Blot

Journal: iScience

Article Title: Overcoming MET-mediated resistance in oncogene-driven NSCLC

doi: 10.1016/j.isci.2023.107006

Figure Lengend Snippet:

Article Snippet: TNO155 , Synthesized at the healthcare business of Merck KGaA, Darmstadt, Germany , NA.

Techniques: Recombinant, Synthesized, Hybridization, Software