Journal: iScience

Article Title: Tracing specificity of immune landscape remodeling associated with distinct anticancer treatments

doi: 10.1016/j.isci.2025.112071

Figure Lengend Snippet:

Article Snippet: Another cohort was treated for 10 or 20 days, every day by intraperitoneal injection with Trametinib (1 mg/kg, MEK inhibitor; TargetMol) plus ABT-737 (30 mg/kg, BCL-XL inhibitor; TargetMol), both diluted in Cremophore and NaCl 0.9%.

Techniques: Recombinant, Software

Journal: Journal of Nuclear Medicine

Article Title: Synthesis of the PET Tracer 124 I-Trametinib for MAPK/ERK Kinase Distribution and Resistance Monitoring

doi: 10.2967/jnumed.120.241901

Figure Lengend Snippet: Receptor tyrosine kinase (RTK) pathway containing RAS and BRAF. MEK is found in receptor tyrosine kinase pathway downstream of both KRAS and BRAF. Trametinib is MEK1/2 inhibitor and Food and Drug Administration–approved for BRAFV600E mutant cancers. By inhibiting MEK activity, trametinib can reduce hyperproliferative signaling from KRAS or BRAF mutants, though on-target toxicity is observed even with wild-type KRAS and BRAF cells.

Article Snippet: Imaging Studies Mice injected with 124 I-trametinib were imaged at 0, 1, 3, 6, 12, 24, and 96 h after injection using a Siemens Inveon PET/CT scanner.

Techniques: Mutagenesis, Activity Assay

Journal: Journal of Nuclear Medicine

Article Title: Synthesis of the PET Tracer 124 I-Trametinib for MAPK/ERK Kinase Distribution and Resistance Monitoring

doi: 10.2967/jnumed.120.241901

Figure Lengend Snippet: Synthesis and cell line specificity of radioiodinated trametinib. (A) Synthesis scheme started from trametinib into boronato-pinacol precursor readily available for iodination. Radiosynthesis and purification were complete within 1.5 h, with radiochemical yield of 70% for 124I-trametinib and specific activity greater than 100 GBq/μmol. (B) HPLC purification shows that most radioiodinated product is distinct from boronato-precursor and identical to cold trametinib. (C) Trametinib is type-III MEK allosteric inhibitor designed to reversibly inhibit adenosine triphosphate from phosphorylating MEK1/2. 124I-trametinib addition to cell lines expressing mutant Ras and or mutant Braf reveal nearly 10-fold range of uptake after 2 h, all of which is blocked with 1 μg of cold trametinib. Human KRAS mutant (G12V, G12C, or G12D) or BRAFV600E cell lines retained higher percentages of 124I-trametinib, with double KRAS BRAF mutant having highest uptake, differentiating KRAS and BRAF mutant tissue from normal. Murine cancer cell lines were all similarly avid for 124I-trametinib except Raw264.7, representing normal KRAS and BRAF and having lower uptake. DMSO = dimethyl sulfoxide; RCY = radiochemical yield; SA = specific activity; UV/Vis = ultraviolet/visible light.

Article Snippet: Imaging Studies Mice injected with 124 I-trametinib were imaged at 0, 1, 3, 6, 12, 24, and 96 h after injection using a Siemens Inveon PET/CT scanner.

Techniques: Purification, Activity Assay, Expressing, Mutagenesis

Journal: Journal of Nuclear Medicine

Article Title: Synthesis of the PET Tracer 124 I-Trametinib for MAPK/ERK Kinase Distribution and Resistance Monitoring

doi: 10.2967/jnumed.120.241901

Figure Lengend Snippet: In vivo distribution of 124I-trametinib in healthy mice primarily in liver, with subsequent elimination through gastrointestinal tract. (A) PET imaging at 1, 3, 6, and 24 h shows broad distribution in tissues, with highest uptake in liver, spleen, and kidneys and later gastrointestinal clearance. Scale bar values in injected dose per gram (%ID/g). (B) Terminal biodistributions at up to 12 h show high heart and lung uptake (5 %ID/g) compared with blood (1 %ID/g) or skin (1–2 %ID/g). Peak uptake in skin was observed at 6 h after injection. GI = gastrointestinal.

Article Snippet: Imaging Studies Mice injected with 124 I-trametinib were imaged at 0, 1, 3, 6, 12, 24, and 96 h after injection using a Siemens Inveon PET/CT scanner.

Techniques: In Vivo, Imaging, Injection

Journal: Journal of Nuclear Medicine

Article Title: Synthesis of the PET Tracer 124 I-Trametinib for MAPK/ERK Kinase Distribution and Resistance Monitoring

doi: 10.2967/jnumed.120.241901

Figure Lengend Snippet: Imaging and biodistribution of 124I-trametinib in mice bearing B16F10 melanomas through 72 h. (A) PET imaging of B16F10 melanoma–bearing mice shows slow tumor uptake through 72 h. Scale bar values in injected dose per gram (%ID/g). (B) Terminal biodistribution reveals maximal uptake of 124I-trametinib in B16F10 tumor footpad between 48 and 72 h after injection. Inset: ratio of %ID/g in tumor to skin, blood, muscle, and heart increased after 24 h, showing retention of 124I-trametinib in tumor relative to other tissues. Tumor uptake after 24 h would allow imaging of naïve tumors in addition to monitoring tumor resistance during conventional trametinib therapy.

Article Snippet: Imaging Studies Mice injected with 124 I-trametinib were imaged at 0, 1, 3, 6, 12, 24, and 96 h after injection using a Siemens Inveon PET/CT scanner.

Techniques: Imaging, Injection

Journal: Journal of Nuclear Medicine

Article Title: Synthesis of the PET Tracer 124 I-Trametinib for MAPK/ERK Kinase Distribution and Resistance Monitoring

doi: 10.2967/jnumed.120.241901

Figure Lengend Snippet: Trametinib therapy reduces uptake of 124I-trametinib systemically. Prior administration of 6 mg kg−1 trametinib intraperitoneally once per day for 3 d shows, by PET, reduced uptake of 124I-trametinib at 24 h (A) and 48 h (B) after administration. Scale bar values in injected dose per gram (%ID/g). (C) Terminal biodistribution on post–48-h PET scan confirms significant decreases in 124I-trametinib uptake in liver, spleen, kidneys, total gastrointestinal tract, bladder, and right-flank B16F10 tumor. Two-way ANOVA was used for biodistribution analysis of 4 mice receiving ×3 trametinib treatment and 5 naïve mice before 124I-trametinib imaging. ****P < 0.001. **P < 0.01. *P < 0.05. LN = lymph node; TGI = total gastrointestinal tract; QD = once daily.

Article Snippet: Imaging Studies Mice injected with 124 I-trametinib were imaged at 0, 1, 3, 6, 12, 24, and 96 h after injection using a Siemens Inveon PET/CT scanner.

Techniques: Injection, Imaging

Journal: bioRxiv

Article Title: SHP2 Inhibition Abrogates MEK inhibitor Resistance in Multiple Cancer Models

doi: 10.1101/307876

Figure Lengend Snippet: A-B , Response of MiaPaCa-2 (A) and Capan-2 (B) subcutaneous xenografts to treatment with SHP099 (75 mg per kg body weight, daily), trametinib (0.25 mg per kg body weight, daily) or both drugs (trametinib 0.25 mg/kg, daily; SHP099 75 mg/kg every other day). Waterfall plot shows response of each tumor after 3 weeks of treatment; n = 8-10 mice per group. (** P < 0.005; *** P < 0.0005, two-tailed Mann Whitney test). C-D , Effects on p-ERK levels in MiaPaCa-2 tumors following 3 weeks of drug treatment, as shown by p-ERK immunoblotting (C) and immunohistochemical analysis (D). E , Syngeneic mice with orthotopic injections of FC1242 (KPC) cells were treated with vehicle, SHP099 (75 mg per kg body weight, daily), trametinib (0.25 mg per kg body weight, daily) or both drugs (trametinib 0.25 mg/kg, daily; SHP099 75 mg/kg every other day), as depicted in the schematic, and tumor size was measured at day 14 (*** P < 0.0005, one-tailed Mann Whitney test) F , Effects on p-ERK activity in FC1242-derived tumors from E. G , Representative haematoxylin and eosin (H&E) stains of drug-treated tumors from E. H , Sizes of tumors from syngeneic mice (C57BL/6J) and athymic nude ( nu/nu ) mice injected orthotopically with the same number of FC1242 (KPC) cells and treated with SHP099 (75 mg per kg body weight, daily), trametinib (1.0 mg per kg body weight, daily) or both drugs (trametinib 1.0 mg/kg + SHP099 75 mg/kg every other day), as depicted in the schematic, measured at day 22 (** P < 0.005, *** P < 0.0005, one-tailed Mann Whitney test). I , Note larger effects seen in immune-competent mice (* P < 0.05, ** P < 0.001; two-sided t test). Numbers under blots indicate relative intensities, compared with untreated controls, quantified by LICOR.

Article Snippet: Selumetinib-AZD6244 (S1008), UO126 (S1102) and trametinib (GSK1120212-S2673) were purchased from Selleckchem.

Techniques: Two Tailed Test, MANN-WHITNEY, Western Blot, Immunohistochemical staining, One-tailed Test, Activity Assay, Derivative Assay, Injection

Journal: bioRxiv

Article Title: SHP2 Inhibition Abrogates MEK inhibitor Resistance in Multiple Cancer Models

doi: 10.1101/307876

Figure Lengend Snippet: A-C , TNBC (A) and HGSC (B) cell lines were treated with DMSO, SHP099, MEK-I (U0126 or AZD6244), or both (COMBO). PrestoBlue (A) and proliferation (B) assays were performed at one week. Colony formation (C) assay was performed at two weeks. Representative results from a minimum of three biological replicates are shown per condition: SHP099 10 μM, U0126 (10 μM) or AZD6244 (1 μM) as indicated, Combo=SHP099 10 μM + UO126 (10 μM)/AZD6244 1 μM (* P < 0.05, ** P < 0.001, *** P < 0.0001, two-sided t test). Red asterisk indicates synergistic interaction between the two drugs by BLISS independent analysis. D , GST-RBD pull down assay from TNBC and HGSC cell line lysates treated with DMSO, SHP099 10 μM, AZD6244 1 μM, or both for 48 h. Image is representative of at least two independent experiments. E , Immunoblots of lysates from TNBC and HSGS cell lines, treated as indicated. Image is representative of three independent experiments. F , ERK-dependent gene expression ( ETV1,4, 5 and FOSL-1 ), assessed by qRT-PCR, in TNBC and HGSC lines treated for 48h with the indicated drugs. G-H , MDA-MB-468 (G) and PDX-2555 (B) mammary fat pad xenografts following treatment with SHP099 (75 mg per kg body weight, daily), trametinib (0.25 mg per kg body weight, daily) or both drugs (trametinib 0.25 mg/kg, daily; SHP099 75 mg/kg every other day). Waterfall representations of tumor responses after 3 weeks of treatment are shown; n = 8-10 mice per group. (** P < 0.005, *** P < 0.0005, two-tailed Mann Whitney test). Numbers under blots indicate relative intensities, compared with untreated controls, quantified by LICOR.

Article Snippet: Selumetinib-AZD6244 (S1008), UO126 (S1102) and trametinib (GSK1120212-S2673) were purchased from Selleckchem.

Techniques: Pull Down Assay, Western Blot, Gene Expression, Quantitative RT-PCR, Two Tailed Test, MANN-WHITNEY

Journal: Nature medicine

Article Title: Genomic profiling informs diagnoses and treatment in vascular anomalies

doi: 10.1038/s41591-023-02364-x

Figure Lengend Snippet: Transduction of BRAF-F486S (a) or RAF1-T145P (c) significantly increased the level of p-ERKs and Trametinib treatment led to a significant reduction of p-ERKs. Three-dimensional lymphatic spheroid sprouting assay showed elevated sprouting activity in HDLECs expressing BRAF-F486S (b) or RAF1-T145P (d) compared to its WT as measured by both cumulative sprout length and number of sprouts. Trametinib treatment led to a significant reduction of both cumulative sprout length and number of sprouts. For a-d, three independent experiments were performed. P values were calculated using 2-sided t-tests and included in each panel (degree of freedom was included in the Source Data for Extended Data Fig. 1), and corrected for multiple testing using the FDR (Benjamini and Hochberg) method. Bar graphs in panels a and c represent mean fold change in the ratio of pERK to GAPDH, normalized to the untreated wild type. Error bars represent standard deviations. In the box and whisker plots in panels b and d, the line in the middle of the box represents the medians, tops and bottoms of the boxes represent the 25th and 75th quartiles respectively, and the whiskers extend to 1.5 times the interquartile range beyond the 25th and 75th quartiles. All the data points that are summarized by the boxplots are superimposed as dots on the plots; minima and maxima can be determined by the highest and lowest dots. e, e’, Expression of KRASWT had no impact on lymphatic vessel morphology in trunk. f, f’, Expression of KRASA146T resulted in lymphatic tissue expansion (asterisks) and dilation of the thoracic duct (dotted line). Red: Expression of transgene in trunk of zebrafish at 5dpf, Green: mrc1a:GFF labeling lymphatic vessels. g, Quantitation of WT EK larvae that were assayed for pericardial edema at 5 dpf. Injected embryos were screened for transgenic mCherry expression in endothelial cells prior to quantitation so that only transgenic expressing embryos were counted. The p-values in the graph were calculated via Fisher’s Exact tests (two-sided) between samples as indicated, followed by Bonferroni correction. Indicated n are the total number of larvae assayed per condition.

Article Snippet: Primary human dermal lymphatic endothelial cells acquired from Promocell (catalog number C-12216; from juvenile foreskin) were retrovirally transduced with wild type or different variants and cultured for 48–72 h. Transduced cells were plated in a 96-well plate (20,000 cells per well) in the presence of carrier (0.1% DMSO) or 300 nM trametinib and cultured for 24 h. Cells were lysed, lysates were cleared by centrifugation and analyzed by SDS–PAGE and western blotting with pERK (Cell Signaling Technology no. 4376) and GAPDH (Santa Cruz Biotechnology no. sc-47724).

Techniques: Transduction, Activity Assay, Expressing, Whisker Assay, Labeling, Quantitation Assay, Injection, Transgenic Assay

Journal: Nature medicine

Article Title: Genomic profiling informs diagnoses and treatment in vascular anomalies

doi: 10.1038/s41591-023-02364-x

Figure Lengend Snippet: a, Of the participants in the study, 44% (69) were already on medical therapy or planned to initiate medical therapy, and 56% (87) were not on medical therapy. b, Distribution of the impact of the molecular finding on medical therapy. The molecular finding supported the chosen therapy in 25 participants and did not support the medication in 1 participant. The molecular diagnosis led to a change in therapy in 13 participants and 4 additional participants planned to change therapy based on the finding. Twelve participants commenced therapy based on the genetic diagnosis and 14 more are planning to initiate therapy. c, Response to therapy. Three participants had worsening of disease, 9 participants had stable disease, 35 participants had improvement in disease and 3 participants had a mixed response to disease. For five participants, it was too early to assess disease response or information was not available (not applicable, NA). d, D-dimer decreased from 37 mg L−1 FEU at the start (off graph) to below 5 mg L−1 FEU within two cycles (CVA14). e, Pulmonary function tests show an increase in forced vital capacity (FVC), forced residual capacity (FRC), forced expiratory volume (FEV1) and total lung capacity (TLC). f–h, MR images before and after trametinib treatment. DCMRL (f) and T2 space (g) demonstrate significant mediastinal perfusion (f, thin arrow) and thickening (g, thin arrow) as well as pleural effusions (g, thick arrow). These findings are significantly reduced post treatment (h, arrows).

Article Snippet: Primary human dermal lymphatic endothelial cells acquired from Promocell (catalog number C-12216; from juvenile foreskin) were retrovirally transduced with wild type or different variants and cultured for 48–72 h. Transduced cells were plated in a 96-well plate (20,000 cells per well) in the presence of carrier (0.1% DMSO) or 300 nM trametinib and cultured for 24 h. Cells were lysed, lysates were cleared by centrifugation and analyzed by SDS–PAGE and western blotting with pERK (Cell Signaling Technology no. 4376) and GAPDH (Santa Cruz Biotechnology no. sc-47724).

Techniques: Biomarker Discovery

Journal: Viruses

Article Title: Enhancing Therapeutic Efficacy of Oncolytic Herpes Simplex Virus with MEK Inhibitor Trametinib in Some BRAF or KRAS-Mutated Colorectal or Lung Carcinoma Models

doi: 10.3390/v13091758

Figure Lengend Snippet: MEK inhibitor (MEKi) Trametinib treatment promoted oHSV replication and tumor cell killing in BRAF V600E-mutated tumor cells. ( A ) Caco-2, Widr, and HT29 cells were pretreated with 15 nM of Trametinib (MEKi) for 2 h and then infected with T1012G at 0.01 PFU/cell. The infected cell pellets were harvested at 48 and 72 h post-infection, respectively. The titration was measured by conventional plaque assay on Vero cells after three freeze–thaw cycles. ( B ) The tumor cell killing activity was assessed using a CCK8 assay. Caco-2, Widr, and HT29 cells were mock treated or pretreated with 15 nM of Trametinib (MEKi) for 2 h and then infected with T1012G at an indicated MOI or MEKi only treated. Cell viability of infected cells was measured with a CCK8 assay (mean ± SD) 48 h after infection. The assay was performed in triplicate. N.S. (not significant); p > 0.05, * p < 0.05.

Article Snippet: MEK inhibitor (MEKi) Trametinib (MCE) was added after the cells had adhered overnight at a final concentration of 15 nM for 2 h. Then, cells were infected with T1012G for 48 h after pretreatment with MEKi.

Techniques: Infection, Titration, Plaque Assay, Activity Assay, CCK-8 Assay

Journal: Viruses

Article Title: Enhancing Therapeutic Efficacy of Oncolytic Herpes Simplex Virus with MEK Inhibitor Trametinib in Some BRAF or KRAS-Mutated Colorectal or Lung Carcinoma Models

doi: 10.3390/v13091758

Figure Lengend Snippet: Down-regulation of STAT1 and PKR phosphorylation when combination of oHSV and MEKi in BRAF mutant cancer cells. ( A ) Caco-2, Widr, and HT29 cells were pretreated with 0.25 μM of Trametinib (MEKi) for 2 h and then mock infected or infected with 0.1 PFU/cell T1012G. The cell pellets were harvested at 24 h post-infection, respectively, and lysates were analyzed by Western blotting for the indicated proteins. ( B – G ). Quantification of the protein level of pERK, ERK, pSTAT1, STAT1, pPKR, and PKR. N.S. (not significant); p > 0.05, * p < 0.05, ** p < 0.01.

Article Snippet: MEK inhibitor (MEKi) Trametinib (MCE) was added after the cells had adhered overnight at a final concentration of 15 nM for 2 h. Then, cells were infected with T1012G for 48 h after pretreatment with MEKi.

Techniques: Mutagenesis, Infection, Western Blot

Journal: Viruses

Article Title: Enhancing Therapeutic Efficacy of Oncolytic Herpes Simplex Virus with MEK Inhibitor Trametinib in Some BRAF or KRAS-Mutated Colorectal or Lung Carcinoma Models

doi: 10.3390/v13091758

Figure Lengend Snippet: MEKi treatment increased oHSV virus yields in KRAS mutated cancer cells. 4T1 ( A ), Pan02 ( B ), LLC ( C ), and CT26 ( D ) cells were pretreated with 0.25 μM of Trametinib (MEKi) for 2 h and then infected with 0.1 PFU/cell T3855. The virus-containing samples were harvested at 24 and 48 h post-infection, respectively. Additionally, then the titration was measured by conventional plaque assay on Vero cells after three freeze–thaw cycles. N.S. (not significant); p > 0.05, * p < 0.05, *** p < 0.001.

Article Snippet: MEK inhibitor (MEKi) Trametinib (MCE) was added after the cells had adhered overnight at a final concentration of 15 nM for 2 h. Then, cells were infected with T1012G for 48 h after pretreatment with MEKi.

Techniques: Virus, Infection, Titration, Plaque Assay

Journal: Viruses

Article Title: Enhancing Therapeutic Efficacy of Oncolytic Herpes Simplex Virus with MEK Inhibitor Trametinib in Some BRAF or KRAS-Mutated Colorectal or Lung Carcinoma Models

doi: 10.3390/v13091758

Figure Lengend Snippet: Decreased STAT1 and PKR expression in MEKi treated KRAS mutated cancer cells. 4T1 and CT26 cells were mock treated or pretreated with 0.25 μM of Trametinib (MEKi) for 2 h and then mock infected or infected with 0.1 PFU/cell T1012G or T3855. The cell pellet was harvested at 24 h post-infection, respectively. The total RNAs were extracted and 0.5 μg of RNAs were reverse transcribed to cDNA as described in Materials and Methods. The PKR and STAT1 mRNAs were quantified and normalized with respect to 18S rRNA and shown as fold change compared with mRNA from mock treated and infected cells (Panel ( A – D )). With the same treatment, cell pellets were harvested and the proteins were electrophoretically separated in a 10% denaturing gel and reacted with indicated antibodies (Panel ( E )). Quantification of the protein level of pERK in 4T1 and CT26 cells (Panel ( F )). N.S. (not significant); p > 0.05, * p < 0.05, ** p < 0.01.

Article Snippet: MEK inhibitor (MEKi) Trametinib (MCE) was added after the cells had adhered overnight at a final concentration of 15 nM for 2 h. Then, cells were infected with T1012G for 48 h after pretreatment with MEKi.

Techniques: Expressing, Infection, Reverse Transcription

Journal: Viruses

Article Title: Enhancing Therapeutic Efficacy of Oncolytic Herpes Simplex Virus with MEK Inhibitor Trametinib in Some BRAF or KRAS-Mutated Colorectal or Lung Carcinoma Models

doi: 10.3390/v13091758

Figure Lengend Snippet: Combined treatment with MEKi Trametinib and oHSV T3855 enhanced antitumor therapeutic activity in LLC and CT26 tumor models. 4T1 (Panel ( A )) or CT26 (Panel ( C )) tumor cells were injected s.c. into the right flanks of Balb/c mice, respectively. LLC (Panel ( B )) tumor cells were injected s.c. into the right flanks of C57BL/6 mice. Different Tumor models averaging 80 mm 3 ( n = 8 per group) were treated via intratumoral injection with PBS or T3855 (1 × 10 7 PFU/animal for 4T1 or LLC; 5 × 10 6 PFU /animal for CT26) on days 1, 8, 15, and 22 and MEKi (trametinib; 1 mg/kg) or vehicle was given from day 1 to 14 via oral gavage. Tumor volumes are shown as mean ± SEM of 8 animals in each group. The complete tumor eradication (Panel ( D )) was summarized at the end of the experiment. N.S. (not significant); p > 0.05, * p < 0.05, ** p < 0.01.

Article Snippet: MEK inhibitor (MEKi) Trametinib (MCE) was added after the cells had adhered overnight at a final concentration of 15 nM for 2 h. Then, cells were infected with T1012G for 48 h after pretreatment with MEKi.

Techniques: Activity Assay, Injection

Journal: Developmental cell

Article Title: Sphingosine 1-phosphate receptor signaling establishes AP-1 gradients to allow for retinal endothelial cell specialization

doi: 10.1016/j.devcel.2020.01.016

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: For MEK inhibition, P6 neonates were subcutaneously injected with 1 mg/kg of Trametinib (GSK1120212, AdooQ Bioscience, CAT#A11029, 0.1 mg/mL solution) or vehicle (1% DMSO in corn oil).

Techniques: Purification, Control, Recombinant, Agarose Gel Electrophoresis, SYBR Green Assay, Staining, Imaging, Sequencing, RNA Sequencing, Software, Microscopy, Real-time Polymerase Chain Reaction, Injection

Journal: Developmental cell

Article Title: Sphingosine 1-phosphate receptor signaling establishes AP-1 gradients to allow for retinal endothelial cell specialization

doi: 10.1016/j.devcel.2020.01.016

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: For pharmacological inhibition of S1PR1, neonates were intragastrically injected with 25 mg/kg of Trametinib NIBR-0213 (Aobious, CAT#AOB1113) or vehicle (DMSO).

Techniques: Purification, Control, Recombinant, Agarose Gel Electrophoresis, SYBR Green Assay, Staining, Imaging, Sequencing, RNA Sequencing, Software, Microscopy, Real-time Polymerase Chain Reaction, Injection