mrtx1257  (MedChemExpress)

Journal: Cell Reports Medicine

Article Title: Developing a therapeutic elastase that stimulates anti-tumor immunity by selectively killing cancer cells

doi: 10.1016/j.xcrm.2025.102446

Figure Lengend Snippet: N17350 broadly kills cancer cells while preserving immune cells (A and B) Various cancer cell lines and primary cancer cells from OvCa patients were treated with increasing doses of N17350 for 24 h, and the EC50 was determined (n = 2–6/dose). (A) Representative killing curves for OvCa patient cancer cells (left), lung cancer cells with distinct KRAS variants (middle), and other tumor types (right). (B) Summary of EC50 values (left) and cancer-cell-type origin (right). (C) Representative tumor growth curve for NCI-H358 and NCI-H2122 tumors (non-small cell lung cancer) treated with N17350 (400 μg/100 mm 3 , intra-tumoral), MRTX1257 (100 mg/kg, oral, daily), or carboplatin (100 mg/kg, intraperitoneal, days 0 and 7); n = 5 mice/group. (D) N17350 efficacy 72 h after a single intra-tumoral injection across various xenograft models; n = 5–7 mice/group. (E) Cancer and non-cancer cells were isolated from OvCa patients, treated with N17350 (500 nM), doxorubicin (10 μM), or oxaliplatin (100 μM) for 24 h, and cell viability was assessed; n = 2–3/patient, n = 31–39 patients/group. (F) Representative N17350 killing curve (left) and eXEC50 values (right) for cancer cells and CD45 + immune cells isolated from the same tumor of OvCa patients ( n = 3/dose across patients). EC50 values for immune cells were assigned as >10,000 nM, as the maximal tested dose failed to achieve a response plateau, precluding accurate curve fitting. ∗ p < 0.05, two-way ANOVA (C and E), Student’s t test: unpaired (D) and paired (F). Results are mean ± SEM. Arrows indicate N17350 treatment. All replicates are independent biological replicates.

Article Snippet: MRTX1257 , MedChemExpress , Cat#2206736-04-9 Lot#49910.

Techniques: Preserving, Injection, Isolation

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

Article Title: Defining the KRAS- and ERK-dependent transcriptome in KRAS-mutant cancers

doi: 10.1126/science.adk0775

Figure Lengend Snippet: (A) KRAS-dependent gene expression changes upon acute (24 hours) KRAS suppression in eight KRAS-mutant PDAC cell lines transiently transfected with KRAS or control non-specific (NS) siRNA. Enrichment of Hallmark KRAS signaling gene sets is shown below. The 677 KRAS-dependent (UP) and 1,051 KRAS-suppressed (DN) genes (log2 fold-change/FC > 0.5, adj. p-val. < 0.05) are indicated by blue and red shaded circles, respectively. The top 200 KRAS-dependent (UP) and KRAS-inhibited (DN) genes comprising the PDAC KRAS UP/DN signatures are indicated by the dotted outlines. (B) Venn diagram indicates the overlap of differentially expressed genes (with unique Entrez gene IDs) upon KRAS siRNA treatment (refer to blue/red shading in (A)) compared to Hallmark KRAS signaling genes. N = 8 (cell lines as biological replicates) for each treatment and control. (C) GSEA for 50 Hallmark gene sets within DE genes following KRAS siRNA treatment. Detection is based on presence of a gene in all 8 PDAC cell lines at >5 reads. NES, normalized enrichment score. (D) KRAS-G12Ci (MRTX1257, 20 nM) induced expression changes summarized over 24 hours and three cell lines (PDAC, CRC, and NSCLC). Genes from siKRAS experiment shown as blue/red boxes in (A) are highlighted with blue/red and indicated by barcode plot below. Enrichment scores (ES) for top 200 UP/DN genes are indicated. (E) GSEA for 50 Hallmark gene sets within RNA-seq data from the indicated human cancer cell line-derived (CDX) or patient-derived (PDX) xenograft tumor samples. CDX mice were treated orally (24 hours) with either KRASG12C or KRASG12D selective inhibitors (G12Ci adagrasib or G12Di MRTX1133, respectively) or vehicle control. PDX mice were treated (21 days) with G12Ci sotorasib alone or together with the EGFRi panitumumab. The top 200 genes from the PDAC KRAS UP/DN gene sets (panel A) were also evaluated. N = 3 for each cell line/treatment combination except HPAC (control) and LS-180 (G12Di), where n = 6.

Article Snippet: The KRAS G12C -selective inhibitor MRTX1257 and the KRAS G12D -selective inhibitor MRTX1133 were synthesized at WuXi AppTec (Wuhan, China) or purchased from Selleckchem (E1051).

Techniques: Gene Expression, Mutagenesis, Transfection, Control, Expressing, RNA Sequencing, Derivative Assay

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

Article Title: Defining the KRAS- and ERK-dependent transcriptome in KRAS-mutant cancers

doi: 10.1126/science.adk0775

Figure Lengend Snippet: (A) Immunoblot analysis of ERK activity in PDAC cell lines stably infected with control vector (Luc) or constitutively activated MEK1 (MEK1-DD), treated with vehicle (DMSO) or indicated inhibitors of each level of the RAF-MEK-ERK cascade (G12Ci/G12Di, MRTX1257/MRTX1133 (20 nM); RAFi, LY3009120 (600 nM); MEKi, trametinib (6 nM); ERKi, SCH772984 (600 nM). Images are representative of 2-3 biological replicates. (B) Growth of PDAC cells stably infected with control vector (Luc) or activated MEK1-DD and treated with the indicated inhibitors. Error bars indicate SE of the mean with 3-4 biological replicates, each with three technical replicates. (C) Immunoblot analysis of ERK and AKT phosphorylation in PDAC cells stably infected with control vector (Luc) or constitutively activated AKT (myr-AKT), treated with G12Ci/G12Di (MRTX1257/MRTX1133, 20 nM) or ERKi (SCH772984, 600 nM). Representative of 2-3 biological replicates. (D) Growth of PDAC cells expressing activated AKT or control vector (Luc) and treated with KRAS G12C/D inhibitors. Error bars indicate SE of the mean with 3-4 biological replicates, each with 3 technical replicates. (E) Differential gene expression analysis for seven PDAC cell lines subjected to ERKi (SCH772984, 1000 nM) treatment for 24 hours versus paired untreated control cells. (F) Top 200 UP/DN genes from 24 hours KRAS siRNA and ERKi RNA-seq experiments are shown as filled blue (UP) or red (DN) circles, with a positive predictive value (agreement/total) for logFC = 0.84. Genes with strongest logFC values (n = 40) are labeled. (G) Evaluation of DE genes following 100 nM treatment of Pa16C PDAC cells with KRASi-G12Di (MRTX1133) or ERKi (SCH772984) for 24 hours (each condition, n = 2). Colored points are DE genes in either treatment (FDR < 0.05); red, log2FC > 0 with either treatment; blue, log2FC < 0 with either treatment; tan, log2FC opposite directions between treatments. Coordinate labels indicate points outside of plotting range. Barcode plots below and left represent log2FC for “median rank KRAS-ERK” signature genes within each experiment and ssGSEA enrichment statistics. (H) GSEA for PDAC KRAS UP/DN, PDAC KRAS-ERK UP/DN, and KRAS-ERK UP/DN based on median rank, along with Hallmark KRAS Signaling signatures in CDX and PDX models treated with G12Di, G12Ci or G12Ci+EGFRi with replicates as described in Fig. 1E. Inhibitors used in (H): G12Ci (CDX), adagrasib; G12Di, MRTX1133; G12Ci (PDX), sotorasib; EGFRi, panitumumab.

Article Snippet: The KRAS G12C -selective inhibitor MRTX1257 and the KRAS G12D -selective inhibitor MRTX1133 were synthesized at WuXi AppTec (Wuhan, China) or purchased from Selleckchem (E1051).

Techniques: Western Blot, Activity Assay, Stable Transfection, Infection, Control, Plasmid Preparation, Phospho-proteomics, Expressing, Gene Expression, RNA Sequencing, Labeling

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

Article Title: Defining the KRAS- and ERK-dependent transcriptome in KRAS-mutant cancers

doi: 10.1126/science.adk0775

Figure Lengend Snippet: (A) KRAS-dependent gene expression changes upon acute (24 hours) KRAS suppression in eight KRAS-mutant PDAC cell lines transiently transfected with KRAS or control non-specific (NS) siRNA. Enrichment of Hallmark KRAS signaling gene sets is shown below. The 677 KRAS-dependent (UP) and 1,051 KRAS-suppressed (DN) genes (log2 fold-change/FC > 0.5, adj. p-val. < 0.05) are indicated by blue and red shaded circles, respectively. The top 200 KRAS-dependent (UP) and KRAS-inhibited (DN) genes comprising the PDAC KRAS UP/DN signatures are indicated by the dotted outlines. (B) Venn diagram indicates the overlap of differentially expressed genes (with unique Entrez gene IDs) upon KRAS siRNA treatment (refer to blue/red shading in (A)) compared to Hallmark KRAS signaling genes. N = 8 (cell lines as biological replicates) for each treatment and control. (C) GSEA for 50 Hallmark gene sets within DE genes following KRAS siRNA treatment. Detection is based on presence of a gene in all 8 PDAC cell lines at >5 reads. NES, normalized enrichment score. (D) KRAS-G12Ci (MRTX1257, 20 nM) induced expression changes summarized over 24 hours and three cell lines (PDAC, CRC, and NSCLC). Genes from siKRAS experiment shown as blue/red boxes in (A) are highlighted with blue/red and indicated by barcode plot below. Enrichment scores (ES) for top 200 UP/DN genes are indicated. (E) GSEA for 50 Hallmark gene sets within RNA-seq data from the indicated human cancer cell line-derived (CDX) or patient-derived (PDX) xenograft tumor samples. CDX mice were treated orally (24 hours) with either KRASG12C or KRASG12D selective inhibitors (G12Ci adagrasib or G12Di MRTX1133, respectively) or vehicle control. PDX mice were treated (21 days) with G12Ci sotorasib alone or together with the EGFRi panitumumab. The top 200 genes from the PDAC KRAS UP/DN gene sets (panel A) were also evaluated. N = 3 for each cell line/treatment combination except HPAC (control) and LS-180 (G12Di), where n = 6.

Article Snippet: The KRAS G12C -selective inhibitor MRTX1257 and the KRAS G12D -selective inhibitor MRTX1133 were synthesized at WuXi AppTec (Wuhan, China) or purchased from Selleckchem (E1051).

Techniques: Gene Expression, Mutagenesis, Transfection, Control, Expressing, RNA Sequencing, Derivative Assay

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

Article Title: Defining the KRAS- and ERK-dependent transcriptome in KRAS-mutant cancers

doi: 10.1126/science.adk0775

Figure Lengend Snippet: (A) Immunoblot analysis of ERK activity in PDAC cell lines stably infected with control vector (Luc) or constitutively activated MEK1 (MEK1-DD), treated with vehicle (DMSO) or indicated inhibitors of each level of the RAF-MEK-ERK cascade (G12Ci/G12Di, MRTX1257/MRTX1133 (20 nM); RAFi, LY3009120 (600 nM); MEKi, trametinib (6 nM); ERKi, SCH772984 (600 nM). Images are representative of 2-3 biological replicates. (B) Growth of PDAC cells stably infected with control vector (Luc) or activated MEK1-DD and treated with the indicated inhibitors. Error bars indicate SE of the mean with 3-4 biological replicates, each with three technical replicates. (C) Immunoblot analysis of ERK and AKT phosphorylation in PDAC cells stably infected with control vector (Luc) or constitutively activated AKT (myr-AKT), treated with G12Ci/G12Di (MRTX1257/MRTX1133, 20 nM) or ERKi (SCH772984, 600 nM). Representative of 2-3 biological replicates. (D) Growth of PDAC cells expressing activated AKT or control vector (Luc) and treated with KRAS G12C/D inhibitors. Error bars indicate SE of the mean with 3-4 biological replicates, each with 3 technical replicates. (E) Differential gene expression analysis for seven PDAC cell lines subjected to ERKi (SCH772984, 1000 nM) treatment for 24 hours versus paired untreated control cells. (F) Top 200 UP/DN genes from 24 hours KRAS siRNA and ERKi RNA-seq experiments are shown as filled blue (UP) or red (DN) circles, with a positive predictive value (agreement/total) for logFC = 0.84. Genes with strongest logFC values (n = 40) are labeled. (G) Evaluation of DE genes following 100 nM treatment of Pa16C PDAC cells with KRASi-G12Di (MRTX1133) or ERKi (SCH772984) for 24 hours (each condition, n = 2). Colored points are DE genes in either treatment (FDR < 0.05); red, log2FC > 0 with either treatment; blue, log2FC < 0 with either treatment; tan, log2FC opposite directions between treatments. Coordinate labels indicate points outside of plotting range. Barcode plots below and left represent log2FC for “median rank KRAS-ERK” signature genes within each experiment and ssGSEA enrichment statistics. (H) GSEA for PDAC KRAS UP/DN, PDAC KRAS-ERK UP/DN, and KRAS-ERK UP/DN based on median rank, along with Hallmark KRAS Signaling signatures in CDX and PDX models treated with G12Di, G12Ci or G12Ci+EGFRi with replicates as described in Fig. 1E. Inhibitors used in (H): G12Ci (CDX), adagrasib; G12Di, MRTX1133; G12Ci (PDX), sotorasib; EGFRi, panitumumab.

Article Snippet: The KRAS G12C -selective inhibitor MRTX1257 and the KRAS G12D -selective inhibitor MRTX1133 were synthesized at WuXi AppTec (Wuhan, China) or purchased from Selleckchem (E1051).

Techniques: Western Blot, Activity Assay, Stable Transfection, Infection, Control, Plasmid Preparation, Phospho-proteomics, Expressing, Gene Expression, RNA Sequencing, Labeling

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

Article Title: Determining the ERK-regulated phosphoproteome driving KRAS-mutant cancer

doi: 10.1126/science.adk0850

Figure Lengend Snippet: (A) Crystal structure of ERK2 bound to the ATP analogue ANP; activating ERK2 mutations (and analogous ERK1 mutations) and the TEY activation loop are shown (PDB: 4NIF). (B) Immunoblots showing mutant ERK expression and rescue of downstream signaling in Pa16C, MIA PaCa-2, and UM53 cells with doxycycline (Dox)-inducible activated ERK1SD (R84S/S170D) or ERK2SD (R67S/S153D) treated with the KRASG12D (MRTX1133, 20 nM) or KRASG12C (MRTX1257, 20 nM) inhibitor for 24 hours. Dox (1 μg/mL) was added 24 hours prior to drug treatments. Blots are representative of three or more independent experiments. Quantification of pRSK and MYC are shown in fig. S1C. (C) Cell viability assays of Pa16C, MIA PaCa-2, UM53 cells with Dox-inducible expression of activated ERK1SD (R84S/S170D) or ERK2SD (R67S/S153D) in response to KRASG12C/D (MRTX1257, MRTX1133) or ERK1/2 inhibitor (SCH772984). Dox treatment (1 μg/mL) was added 24 hours prior to drug treatments. Graphs show the mean percent growth of three replicates +/− standard error of the mean. (D) RPPA Spearman correlation coefficients comparing log2(FC) of signals from 147 antibodies for the individual six cell lines and all cell lines considered together. Cells were treated with siRNAs targeting NS (non-specific control), KRAS, MAPK3 (ERK1) and/or MAPK1 (ERK2), for 72 hours as in (fig. S1E). Statistical analysis was performed from four biological replicates. The individual treatments and full protein list can be found in fig. S1G and data S1. (E) RNA transcript expression changes induced by 24-hour KRASG12D inhibition (MRTX1133, 100 nM) versus DMSO control in Pa16C cells expressing activated ERK1SD (cyan), ERK2SD (pink), or GFP control (black). Genes are ranked by expression changes in GFP expressing cells. Dox (1 μg/mL) was added 24 hours prior to drug treatments. Statistical analysis was performed from two biological replicates. Genes shown for ERK1SD and ERK2SD (points) are those with significant DE induced by KRASG12D inhibition in GFP control cells (adj. p-val. < 0.01). (F) Venn diagrams representing the overlap between genes upregulated (G12Di UP) or downregulated (G12Di DN) by KRASG12D inhibition in GFP expressing cells (green; |log2FC| > 0.25 and adj. p-val. < 0.01) and genes rescued by ERK1SD (cyan) and ERK2SD (pink). The center overlap (purple) depicts genes rescued by both ERK1SD and ERK2SD, the genes in only the green area are not rescued by either, in the cyan area rescued by only ERK1SD, and in the purple area by only ERK2SD. The term “rescue” indicates an expression change (log2FC) less extreme than the GFP control but no p-values were used to represent the degree of those expression changes.

Article Snippet: The KRAS G12C -selective inhibitor MRTX1257 was provided by Mirati Therapeutics.

Techniques: Activation Assay, Western Blot, Mutagenesis, Expressing, Control, Inhibition

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

Article Title: Determining the ERK-regulated phosphoproteome driving KRAS-mutant cancer

doi: 10.1126/science.adk0850

Figure Lengend Snippet: (A) Flow diagram of the MIB/MS experiment was created with BioRender.com. (B) MIB/MS analyses of six KRAS mutant cell lines treated for 1 or 24 hours with ERKi (SCH772984, 1 μM). (C) Experimental workflow for proteomics analyses, created with BioRender.com. (D) Differentially expressed phosphosites following 1 and 24 hours ERK inhibition in the same six PDAC cell lines as in (A and B). Phosphosites previously reported in the ERK Compendium are highlighted, and top significantly dysregulated sites identified within the ERK Compendium as direct substrates of ERK are labeled. (E) Overlap between ERK-dependent phosphosites (p < 0.05) or phosphoproteins in PDAC and phosphosites/phosphoproteins reported in the CPTAC PDAC dataset (fdr < 0.05), ERK Compendium, PhosphoSitePlus (PSP) regulatory sites, and Post-Translational Signature Database (PTMsigDB). (F) Differentially expressed phosphosites following 24-hour KRASG12C inhibition (MRTX1257, 20 nM, G12Ci) or RAS(ON) multi-selective tri-complex inhibition (RMC-7977, 10 nM, RASi). ERK-dependent phosphosites (p<0.05, 24 h) are annotated by blue (downregulated) or red (upregulated) points. ERK-regulated phosphosites at 1 and 24 hours are annotated as bars below the volcano plot.

Article Snippet: The KRAS G12C -selective inhibitor MRTX1257 was provided by Mirati Therapeutics.

Techniques: Mutagenesis, Inhibition, Labeling

Journal: Journal of Translational Medicine

Article Title: KRAS G12C inhibition using MRTX1257: a novel radio-sensitizing partner

doi: 10.1186/s12967-023-04619-0

Figure Lengend Snippet: MRTX1257 radio-sensitizes in vitro CT26 and LL2 tumor cell lines according to their KRAS mutational status. Clonogenic survival assays were performed after treatment using RT and MRTX1257 at various doses in different cell lines harboring different KRAS mutational profiles. Normalized survival fractions are represented in mean ± standard-error to mean (SEM), with n = 3 to 6 replicates per condition. Survival curves are extrapolations according to the linear quadratic model. ****: p < 0.0001 (one-way ANOVA). Survival curves for ( A ) CT26 KRAS G12C+/+ ; B CT26 WT; C LL2 NRAS −/− ; D LL2 WT. E, F Dose–response curves for CT26 WT and LL2 WT cell lines exposed to various concentrations of KRAS. G12C inhibitor MRTX1257

Article Snippet: MRTX1257 was provided by Mirati Therapeutics and reconstituted using cyclodextrin Captisol (Ligand, San Diego, USA) in case of oral administration in mice or dimethylsulfoxide (DMSO) for in vitro experiments.

Techniques: In Vitro

Journal: Journal of Translational Medicine

Article Title: KRAS G12C inhibition using MRTX1257: a novel radio-sensitizing partner

doi: 10.1186/s12967-023-04619-0

Figure Lengend Snippet: MRTX1257 increases the efficacy of RT in nude mice bearing CT26 KRAS G12C+/+ however without achieving durable responses A Athymic nude mice were subcutaneously inoculated with CT26 KRAS G12C+/+ cells. Once the tumors reached an average volume of 90–100 mm 3 , mice received via oral administration 50 mg/kg of MRTX1257 or vehicle. The day after, mice received a single fraction of 6 Gy on the tumor mass. MRTX1257 at the dose of 50 mg/kg or vehicle were then administered at D1 and D3 after RT. The experiment was repeated twice. B Tumor growth (mean ± SEM) ***: p < 0.001; ****: p < 0.0001 (two-way ANOVA). C Mean tumor volumes in each group 10 days after RT are represented. *: p < 0.05; ***: p < 0.001; ****: p < 0.0001 (one-way ANOVA). D Individual growth profiles of CT26 KRAS. G12C+/+ tumors are represented for each condition. E Survival Kaplan–Meier curves were compared between each group using the log-rank test. **: p < 0.01; ****: p < 0.0001

Article Snippet: MRTX1257 was provided by Mirati Therapeutics and reconstituted using cyclodextrin Captisol (Ligand, San Diego, USA) in case of oral administration in mice or dimethylsulfoxide (DMSO) for in vitro experiments.

Techniques:

Journal: Journal of Translational Medicine

Article Title: KRAS G12C inhibition using MRTX1257: a novel radio-sensitizing partner

doi: 10.1186/s12967-023-04619-0

Figure Lengend Snippet: MRTX1257 increases the efficacy of RT in immunocompetent BALB/c mice bearing CT26 KRAS G12C+/+ tumors and achieved durable responses in association with RT. A BALB/c mice were subcutaneously inoculated with CT26 KRAS G12C+/+ cells. Once the tumors reached an average volume of 90–100 mm 3 , mice received via oral administration 50 mg/kg of MRTX1257 or vehicle. The day after, mice received a single fraction of 6 Gy on the tumor mass. MRTX1257 at the dose of 50 mg/kg or vehicle were then administered at D1 and D3 after RT. The experiment was repeated twice. B Tumor growth (mean ± SEM). **: p < 0.01; ***: p < 0.001; ****: p < 0.0001 (two-way ANOVA). ( C ) Mean tumor volumes in each group 10 days after RT are represented. **: p < 0.01; ***: p < 0.001; ****: p < 0.0001 (one-way ANOVA). D Individual growth profiles of CT26 KRAS G12C +/+ tumors are represented for each condition. The number of mice achieving durable response with no tumor being assessed at the end of the experiment is indicated below each panel. E Survival Kaplan–Meier curves were compared between each group using the log-rank test. *: p < 0.05; **: p < 0.01; ****: p < 0.0001. F Immunocompetent BALB/c mice bearing CT26 KRAS G12C+/+ tumors and cured by the combination of RT and MRTX1257 were subsequently rechallenged with either 1.10 6 CT26 KRAS G12C+/+ (top) or 5.10 5 CT26 WT cells (bottom) subcutaneously injected in their contralateral flank. Their survivals were then compared with those of naive mice receiving similar injections

Article Snippet: MRTX1257 was provided by Mirati Therapeutics and reconstituted using cyclodextrin Captisol (Ligand, San Diego, USA) in case of oral administration in mice or dimethylsulfoxide (DMSO) for in vitro experiments.

Techniques: Injection

Journal: Journal of Translational Medicine

Article Title: KRAS G12C inhibition using MRTX1257: a novel radio-sensitizing partner

doi: 10.1186/s12967-023-04619-0

Figure Lengend Snippet: MRTX1257 does not have any significant effect alone or in combination with RT in immunocompetent BALB/c mice bearing CT26 WT tumors. A BALB/c mice were subcutaneously inoculated with CT26 WT cells. Once the tumors reached an average volume of 90–100 mm 3 , mice received via oral administration 50 mg/kg of MRTX1257 or vehicle. The day after, mice received a single fraction of 6 Gy on the tumor mass. MRTX1257 at the dose of 50 mg/kg or vehicle were then administered at D1 and D3 after RT. The experiment was repeated three times. B Tumor growth (mean ± SEM). ***: p < 0.001 (two-way ANOVA). C Mean tumor volumes in each group 10 days after RT are represented. **: p < 0.01; ***: p < 0.001 (one-way ANOVA). D Individual growth profiles of CT26 KRAS G12C. +/+ tumors are represented for each condition. The number of mice achieving durable response with no tumor being assessed at the end of the experiment is indicated below each panel. E Survival Kaplan–Meier curves were compared between each group using the log-rank test. ***: p < 0.001

Article Snippet: MRTX1257 was provided by Mirati Therapeutics and reconstituted using cyclodextrin Captisol (Ligand, San Diego, USA) in case of oral administration in mice or dimethylsulfoxide (DMSO) for in vitro experiments.

Techniques:

Journal: Journal of Translational Medicine

Article Title: KRAS G12C inhibition using MRTX1257: a novel radio-sensitizing partner

doi: 10.1186/s12967-023-04619-0

Figure Lengend Snippet: MRTX1257 increases the anti-proliferative effect of RT in CT26 KRAS G12C+/+ tumors but not in CT26 WT tumors. A BALB/c mice were subcutaneously inoculated with CT26 WT cells or CT26 KRAS G12C+/+ cells. Once the tumors reached an average volume of 90–100 mm 3 , mice received via oral administration 50 mg/kg of MRTX1257 or vehicle. The day after, mice received a single fraction of 6 Gy on the tumor mass. MRTX1257 at the dose of 50 mg/kg or vehicle were then administered at D1 and D3 after RT. At D4 after RT, mice were sacrificed, tumors were harvested and then fixated using paraformaldehyde (PFA) 4%. Immunohistochemistry for Ki67 and then the quantification of Ki67 + cells were performed. B Photographs at magnification 13X of 4 µm-thick CT26 KRAS G12C+/+ (left) or CT26 WT (right) tumor slices stained with anti-Ki67 antibody. Two representative photographs, each of them from a different tumor, are represented for each condition. C Quantification of Ki67 + cells per mm 2 in CT26 KRAS G12C +/+ tumor slices. Each point is representative of a single tumor. Mean ± standard-errors to mean (SEM). N = 3–5 tumors/group. *: p < 0.05; **: p < 0.01 (one-way ANOVA). D Quantification of Ki67 + cells per mm. 2 in CT26 WT tumor slices. Each point is representative of a single tumor. Mean ± standard-errors to mean (SEM). n = 3 tumors/group. *: p < 0.05 (one-way ANOVA)

Article Snippet: MRTX1257 was provided by Mirati Therapeutics and reconstituted using cyclodextrin Captisol (Ligand, San Diego, USA) in case of oral administration in mice or dimethylsulfoxide (DMSO) for in vitro experiments.

Techniques: Immunohistochemistry, Staining

Journal: Journal of Translational Medicine

Article Title: KRAS G12C inhibition using MRTX1257: a novel radio-sensitizing partner

doi: 10.1186/s12967-023-04619-0

Figure Lengend Snippet: An increase in tumor-infiltrating CD8 + T cells is not sufficient to explain the radio-sensitizing effect of MRTX1257. A BALB/c mice were subcutaneously inoculated with CT26 WT cells or CT26 KRAS G12C +/+ cells. Once the tumors reached an average volume of 90–100 mm 3 , mice received via oral administration 50 mg/kg of MRTX1257 or vehicle. The day after, mice received a single fraction of 6 Gy on the tumor mass. MRTX1257 at the dose of 50 mg/kg or vehicle were then administered at D1 and D3 after RT. At D4 after RT, mice were sacrificed, tumors were harvested and then fixated using paraformaldehyde (PFA) 4%. Immunohistochemistry analyses for CD8 and then the quantification of CD8 + cells were performed. B Photographs at magnification 1.5X of 4 µm-thick CT26 KRAS G12C+/+ (left) or CT26 WT (right) tumor slices stained with anti-CD8 antibody. C Quantification of CD8 + cells per mm 2 in CT26 KRAS G12C+/+ tumor slices. Each point is representative of a single tumor. Mean ± standard-errors to mean (SEM). n = 3–5 tumors/group. *: p < 0.05 (one-way ANOVA). D Quantification of CD8 + cells per mm 2 in CT26 WT tumor slices. Each point is representative of a single tumor. Mean ± standard-errors to mean (SEM). n = 3 tumors/group. Ns: non-significant (one-way ANOVA)

Article Snippet: MRTX1257 was provided by Mirati Therapeutics and reconstituted using cyclodextrin Captisol (Ligand, San Diego, USA) in case of oral administration in mice or dimethylsulfoxide (DMSO) for in vitro experiments.

Techniques: Immunohistochemistry, Staining

Journal: Journal of Translational Medicine

Article Title: KRAS G12C inhibition using MRTX1257: a novel radio-sensitizing partner

doi: 10.1186/s12967-023-04619-0

Figure Lengend Snippet: RT and MRTX1257 reshape the tumor immune microenvironment in CT26 KRAS G12C+/+ tumors. Flow cytometry analyses were performed in single-suspensions derived from CT26 KRAS G12C+/+ tumors. Results are represented in mean ± standard-error to mean (SEM). Each point represents a single tumor in both the untreated control group (NIR + VEHICLE) and the group treated with RT alone (IR 6 Gy + VEHICLE). Each point represents 2 different tumors pooled in a single sample in both the group treated with MRTX1257 alone (NIR + MRTX1257) and the group treated with combined RT and MRTX1257 (IR 6 Gy + MRTX1257). n = 3/group. Except for dendritic cells, the proportions of the different immune cell subtypes are expressed in percentage in viable cells whereas the expression of PD-L1, major histocompatibility complex II (MHC II), and CD80 are expressed in the difference in mean fluorescence intensity compared to an unstained control within each condition (Delta MFI). NS: non-significant; *: p < 0.05; **: p < 0.01; ***: p < 0.001; ****: p < 0.0001 (one-way ANOVA) A BALB/c mice were subcutaneously inoculated with CT26 KRAS G12C+/+ cells. Once the tumors reached an average volume of 90–100 mm 3 , mice received via oral administration 50 mg/kg of MRTX1257 or vehicle. The day after, mice received a single fraction of 6 Gy on the tumor mass. MRTX1257 at the dose of 50 mg/kg or vehicle were then administered at D1 and D3 after RT. At D4 after RT, mice were sacrificed, tumors were harvested and were used to perform flow cytometry. B Expression of PD-L1 in tumor and stromal CD45 − cells. Mean ± SEM (left panel) and histogram overlay (right panel). C Proportion of lymphoid cells, conventional CD4 + T cells, CD8 + T cells and Tregs. D Proportion of myeloid cells and expression of PD-L1 within myeloid cells. E Proportion of dendritic cells type 2 (cDC2) in percentage of myeloid cells and expression of MHC II within cDC2. F Proportion of inflammatory monocytes and expression of MHC II within inflammatory monocytes. G Proportion of macrophages and expression of CD80 within macrophages

Article Snippet: MRTX1257 was provided by Mirati Therapeutics and reconstituted using cyclodextrin Captisol (Ligand, San Diego, USA) in case of oral administration in mice or dimethylsulfoxide (DMSO) for in vitro experiments.

Techniques: Flow Cytometry, Derivative Assay, Control, Expressing, Immunopeptidomics, Fluorescence