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 Oncology

Article Title: KRAS G 12 C ‐inhibitor‐based combination therapies for pancreatic cancer: insights from drug screening

doi: 10.1002/1878-0261.13725

Figure Lengend Snippet: Efficacy of the TNO155 and Sotorasib combination in a KRAS G12C ‐mutated patient‐derived organoids (PDOs). (A) Case report of a KRAS G12C ‐mutated pancreatic ductal adenocarcinoma (PDAC) patient, from which the patient‐derived organoid (PDO) line PDO‐51T was isolated. After the resection of the tumor (pT3, pN2, cM0, R0, G2) the male patient was treated with adjuvant Gemcitabine/Capecitabine for 4 months. Afterwards, the patient was treated with mFOLFIRINOX and Gemcitabine/nab‐Paclitaxel. The overall survival was 15 months. (B) Results of the panel sequencing of PDO‐51T. (C) The top five enriched and depleted signatures of a single sample Gene Set Enrichment Analysis (ssGSEA) of the PDO‐51T line. (D) The indicated Sotorasib doses were used to determine growth over the indicated time points of PDO‐51T, measured by live cell imaging. n = 6. (E, F) PDO‐51T were treated with 41 n m Sotorasib ( n = 6), 41 n m TNO155 ( n = 6), 41 n m Nintedanib ( n = 3), 40 n m Sotorasib/41 n m TNO155 ( n = 6), and 40 n m Sotorasib/41 n m Nintedanib ( n = 6) or left as vehicle controls ( n = 6). Growth curves were determined by live cell imaging. Yellow scale bar: 400 μ m . (G) Quantification of the growth after 192 h. Data of the experiment described in F were used to calculate the relative growth (ratio treatment groups versus control). One‐way ANOVA with correction for multiple testing according to Tukey: ** P < 0.01, * P < 0.05. Median and quartiles are shown.

Article Snippet: The cherry‐picked drug library (stock 10 m m , dissolved in DMSO), Sotorasib (AMG‐510) (#S8830), Batoprotafib (TNO155) (#S8987), Decitabine (#S1200), Ceralasertib (AZD8738) (#S7693), BI‐3406 (#S8916), MS023 (#S8112), and Nintedanib (#S1010) were purchased from Selleckchem (Cologne, Germany).

Techniques: Derivative Assay, Isolation, Adjuvant, Sequencing, Live Cell Imaging, Control

Journal: Molecular Oncology

Article Title: KRAS G 12 C ‐inhibitor‐based combination therapies for pancreatic cancer: insights from drug screening

doi: 10.1002/1878-0261.13725

Figure Lengend Snippet: Synergism of Sotorasib with TNO155. (A) 2D pancreatic ductal adenocarcinoma (PDAC) cells (51T‐2D) were generated from PDO‐51T. (B) Phospho‐ERK, pan‐ERK, and KRAS western blot 24 h after the treatment of 51T‐2D cells with the indicated doses of Sotorasib. One representative blot out of three replicates ( n = 3) is depicted. (C) Phospho‐ERK Quantification of three independent experiments ( n = 3) corresponding to B. (D, E) Clonogenic growth assay of 51T‐2D cells treated with the indicated doses of TNO155 and Sotorasib. (D) One representative experiment. (E) Quantification of three independent experiments ( n = 3). One‐way ANOVA with Tukey's correction for multiple testing: *** P < 0.001. (F) Bliss synergy score of the quantified clonogenic growth assays of 51T‐2D cells was calculated with the synergy finder platform. (G, H) MiaPaca2 and 51T‐2D cells were treated as indicated. After 72 h the cells were stained with propidium iodide (PI) and cell cycle FACS was performed. Depicted is the ratio of cells in the S‐phase of the cell cycle to cells in the G1‐phase ( n = 3). One‐way ANOVA with correction for multiple testing according to Tukey: * P < 0.05. (I) Staining for Ki67 and cleaved caspase 3 (CC3) in PDO‐51T treated as indicated ( n = 1). Scale bar = 10 μm. (J) Phospho‐ERK, pan‐ERK, and KRAS western blot 72 h after the treatment of 51T‐2D cells with the indicated doses of Sotorasib and/or TNO155. One representative blot out of three replicates ( n = 3) is depicted. (K) Quantification of three independent experiments ( n = 3) corresponding to J. One‐way ANOVA with Tukey's correction for multiple testing: * P < 0.05. Mean and the standard deviation (SD) are shown.

Article Snippet: The cherry‐picked drug library (stock 10 m m , dissolved in DMSO), Sotorasib (AMG‐510) (#S8830), Batoprotafib (TNO155) (#S8987), Decitabine (#S1200), Ceralasertib (AZD8738) (#S7693), BI‐3406 (#S8916), MS023 (#S8112), and Nintedanib (#S1010) were purchased from Selleckchem (Cologne, Germany).

Techniques: Generated, Western Blot, Growth Assay, Staining, Standard Deviation

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