Journal: Pharmaceutics

Article Title: Validation of Liquid Chromatography Coupled with Tandem Mass Spectrometry for the Determination of 12 Tyrosine Kinase Inhibitors (TKIs) and Their Application to Therapeutic Drug Monitoring in Adult and Pediatric Populations

doi: 10.3390/pharmaceutics16010005

Figure Lengend Snippet: Characteristics of the 12 TKIs analyzed: tyrosine kinase target, indication in adults, and concentrations reported in the literature (NSCLC: non-small cell lung cancer).

Article Snippet: The reference materials for alectinib, alectinib M4, AZ5104 (osimertinib active metabolite), crizotinib-13C2D5, imatinib, imatinib-D8, and osimertinib were purchased from LGC Standards (Molsheim, France).

Techniques:

Journal: Pharmaceutics

Article Title: Validation of Liquid Chromatography Coupled with Tandem Mass Spectrometry for the Determination of 12 Tyrosine Kinase Inhibitors (TKIs) and Their Application to Therapeutic Drug Monitoring in Adult and Pediatric Populations

doi: 10.3390/pharmaceutics16010005

Figure Lengend Snippet: Concentrations of calibration standards and quality controls (QC) for the three groups of TKIs (ng/mL).

Article Snippet: The reference materials for alectinib, alectinib M4, AZ5104 (osimertinib active metabolite), crizotinib-13C2D5, imatinib, imatinib-D8, and osimertinib were purchased from LGC Standards (Molsheim, France).

Techniques:

Journal: Pharmaceutics

Article Title: Validation of Liquid Chromatography Coupled with Tandem Mass Spectrometry for the Determination of 12 Tyrosine Kinase Inhibitors (TKIs) and Their Application to Therapeutic Drug Monitoring in Adult and Pediatric Populations

doi: 10.3390/pharmaceutics16010005

Figure Lengend Snippet: Retention time (Rt), parent and fragment ions, and collision energies for the studied TKIs and IS.

Article Snippet: The reference materials for alectinib, alectinib M4, AZ5104 (osimertinib active metabolite), crizotinib-13C2D5, imatinib, imatinib-D8, and osimertinib were purchased from LGC Standards (Molsheim, France).

Techniques:

Journal: Pharmaceutics

Article Title: Validation of Liquid Chromatography Coupled with Tandem Mass Spectrometry for the Determination of 12 Tyrosine Kinase Inhibitors (TKIs) and Their Application to Therapeutic Drug Monitoring in Adult and Pediatric Populations

doi: 10.3390/pharmaceutics16010005

Figure Lengend Snippet: Characteristics of TKI-treated patients monitored in our laboratory from 2014 up to June 2023.

Article Snippet: The reference materials for alectinib, alectinib M4, AZ5104 (osimertinib active metabolite), crizotinib-13C2D5, imatinib, imatinib-D8, and osimertinib were purchased from LGC Standards (Molsheim, France).

Techniques:

Journal: Pharmaceutics

Article Title: Validation of Liquid Chromatography Coupled with Tandem Mass Spectrometry for the Determination of 12 Tyrosine Kinase Inhibitors (TKIs) and Their Application to Therapeutic Drug Monitoring in Adult and Pediatric Populations

doi: 10.3390/pharmaceutics16010005

Figure Lengend Snippet: Validation parameters of the 12 TKIs. Normalized matrix effect CV and recovery CV were calculated for the 6 lots of plasma. LOD = limit of detection, LOQ = limit of quantification, and CV = coefficient of variation.

Article Snippet: The reference materials for alectinib, alectinib M4, AZ5104 (osimertinib active metabolite), crizotinib-13C2D5, imatinib, imatinib-D8, and osimertinib were purchased from LGC Standards (Molsheim, France).

Techniques:

Journal: Pharmaceutics

Article Title: Validation of Liquid Chromatography Coupled with Tandem Mass Spectrometry for the Determination of 12 Tyrosine Kinase Inhibitors (TKIs) and Their Application to Therapeutic Drug Monitoring in Adult and Pediatric Populations

doi: 10.3390/pharmaceutics16010005

Figure Lengend Snippet: Descriptive statistics of 10 TKIs for adults and children: median concentrations, interquartile range (IQR), and mean and confidence interval (CI) 95% in ng/mL.

Article Snippet: The reference materials for alectinib, alectinib M4, AZ5104 (osimertinib active metabolite), crizotinib-13C2D5, imatinib, imatinib-D8, and osimertinib were purchased from LGC Standards (Molsheim, France).

Techniques: Concentration Assay

Journal: Cell reports

Article Title: Capicua suppresses YAP1 to limit tumorigenesis and maintain drug sensitivity in human cancer

doi: 10.1016/j.celrep.2022.111443

Figure Lengend Snippet: (A, F, and K) CTG viability curves in HCC1359 (A), H1975 (F), and HCC364 (K) following treatment with trametinib (tram), Osimertinib (osi), or vemurafenib (vem), respectively. (B, G, and L) Relative cell number of HCC1359 (B), H1975 (G), and HCC364 (L) cells expressing sgCtrl, sgCIC1, or sgCIC2 and treated (6 d) with tram, osi, or vem, respectively. **p < 0.01. (C, H, and M) Crystal violet (CV) assay of HCC1359 (C), H1975 (H), and HCC364 (M) cells expressing sgCtrl, sgCIC1, or sgCIC2 treated (10 d) with tram, osi, or vem, respectively. (D, I, and N) Relative cell number of HCC1359 (D), H1975 (I), and HCC364 (N) cells expressing sgCtrl, sgCIC1, or sgCIC2 ± YAP1 KD (shYAP1–2 and shYAP1–3) treated (6 d) with tram, osi, or vem, respectively. *p < 0.05, **p < 0.01. (E, J, and O) CV assay of HCC1359 (E), H1975 (J), and HCC364 (O) cells expressing sgCtrl, sgCIC1, or sgCIC2 ± YAP1 KD (shYAP1–2 and shYAP1–3) treated (10 d) with tram, osi, or vem, respectively. Error bars represent SD in all figures.

Article Snippet: Osimertinib , MedChemExpress , Cat# HY-15772.

Techniques: Expressing

Journal: Cell reports

Article Title: Capicua suppresses YAP1 to limit tumorigenesis and maintain drug sensitivity in human cancer

doi: 10.1016/j.celrep.2022.111443

Figure Lengend Snippet: (A) H1975 xenografts comparing vehicle or osimertinib-treated H1975, H1975 with CIC KO, or H1975 with CIC KO + YAP1 KD (n = 10). (B) Tumor weights from mice harboring H1975, H1975 CIC KO, or H1975 CIC KO + YAP1 KD ± osimertinib. (C) Immunoblots of CIC, YAP, and HSP90 expression in H1975 xenografts derived from tumors of different groups in (A). (D) Representative figures of parental, CIC KO, or CIC KO + YAP1 KD in HCC1359 tumor-bearing mice. (E) Bar graph comparing the incidence of HCC1359 (n = 2/20), HCC1359 CIC KO (n = 13/20), or HCC1359 CIC KO + YAP1 KD (n = 1/20) tumor formation in mice. (F) Relative growth of HCC1359 CIC KO tumors treated with vehicle or verteporfin. (G) Correlation plot between CIC and YAP expression from LC specimens (tau = −0.33, p = 0.0002, n = 100). (H) Representative images of CIC and YAP-stained LC tissue. Scale bar, 50 μm. (I) Model of CIC-mediated repression of YAP1 and phenotypes. Error bars represent SD.

Article Snippet: Osimertinib , MedChemExpress , Cat# HY-15772.

Techniques: Western Blot, Expressing, Derivative Assay, Staining

Journal: Cell reports

Article Title: Capicua suppresses YAP1 to limit tumorigenesis and maintain drug sensitivity in human cancer

doi: 10.1016/j.celrep.2022.111443

Figure Lengend Snippet:

Article Snippet: Osimertinib , MedChemExpress , Cat# HY-15772.

Techniques: Virus, Recombinant, Transfection, Western Blot, Plasmid Preparation, Purification, Mutagenesis, Chromatin Immunoprecipitation, Magnetic Beads, Reporter Assay, Luciferase, Promoter Assay, shRNA, Software, Imaging, Sequencing

Journal: Oncogene

Article Title: RBM15 facilitates osimertinib resistance of lung adenocarcinoma through m6A-dependent epigenetic silencing of SPOCK1

doi: 10.1038/s41388-024-03220-z

Figure Lengend Snippet: A , B Representative image of IHC staining and IHC scores of RBM15 from EGFR wild-type (WT) or mutant patients. Mann–Whitney test. C , D Immunofluorescence of RBM15 protein expression in LUAD PDO with or without EGFR mutations (scale bar: 50 μm). T test. E OS in TCGA LUAD cases with EGFR mutation according to RBM15 expression. F PFS after initiation of osimertinib treatment in patients with EGFR mutations expressing high vs. low RBM15 mRNA levels. G , H Osimertinib IC 50 value of HCC827 and H1975 cells transfected with NC and RBM15-OE. T test. Error bars represent the means ± SDs. * P < 0.05, **** P < 0.0001.

Article Snippet: When the tumors reached a longitudinal diameter of 5 mm, the mice were randomly assigned to each group and subsequently treated orally with an equal volume of osimertinib mesylate (Selleck, S5078, TX, 20 mg/kg/day) or corn oil by gavage every 2 days.

Techniques: Immunohistochemistry, Mutagenesis, MANN-WHITNEY, Immunofluorescence, Expressing, Transfection

Journal: Oncogene

Article Title: RBM15 facilitates osimertinib resistance of lung adenocarcinoma through m6A-dependent epigenetic silencing of SPOCK1

doi: 10.1038/s41388-024-03220-z

Figure Lengend Snippet: A H1975 cells were exposed to increasing concentrations of osimertinib and cultured for 24 weeks to select H1975-OR cells (top panel). H1975 and H1975-OR cells were incubated with osimertinib for 48 h, cell viability was measured using the CCK-8 assay, and the IC 50 was calculated (bottom panel). T test. B Western blot investigation of EGFR, ERK, and AKT protein levels and their phosphorylation. C , D RBM15 protein and mRNA expression levels in H1975 and H1975-OR were assessed using western blot and qRT-PCR. T test. E Osimertinib IC 50 of H1975-OR cells transfected with si-NC, si-RBM15#1, or si-RBM15#2. T test. F A schematic diagram for the construction of drug-resistant organoids. G PDO1-OR and osimertinib-sensitive organoids were incubated with osimertinib for 48 h, cell viability was measured using the CCK-8 assay, and the IC 50 was calculated. T test. H Osimertinib IC 50 value of PDO1-OR transfected with sh-NC or sh-RBM15. T test. I , J Effects of RBM15 knockdown on H1975-OR and PDO1-OR apoptosis assessed using flow cytometry and TUNEL staining. T test. K Western blot analysis of EGFR and AKT proteins and their phosphorylation levels after knockdown of RBM15 and 48-h osimertinib treatment. L Representative images of xenograft tumors from sh-NC, sh-NC + Osi, and sh-RBM15 + Osi groups. M , N Tumor growth curve and tumor weights. T test. O Representative images of Ki-67 positive staining in xenografted tumors. T test. Error bars represent the means ± SDs. * P < 0.05, ** P < 0.01, **** P < 0.0001.

Article Snippet: When the tumors reached a longitudinal diameter of 5 mm, the mice were randomly assigned to each group and subsequently treated orally with an equal volume of osimertinib mesylate (Selleck, S5078, TX, 20 mg/kg/day) or corn oil by gavage every 2 days.

Techniques: Cell Culture, Incubation, CCK-8 Assay, Western Blot, Phospho-proteomics, Expressing, Quantitative RT-PCR, Transfection, Knockdown, Flow Cytometry, TUNEL Assay, Staining

Journal: Oncogene

Article Title: RBM15 facilitates osimertinib resistance of lung adenocarcinoma through m6A-dependent epigenetic silencing of SPOCK1

doi: 10.1038/s41388-024-03220-z

Figure Lengend Snippet: A Venn diagrams were generated from the set of genes enriched for transcripts substantially altered after RBM15 silencing (RNA-seq) and the set of genes enriched for m6A-modified transcripts (m6A-seq). Sixteen genes were chosen based on overlap. B Top motif identified by HOMER with m6A-seq peaks. C Peak regions of m6A modifications detected by MeRIP-seq. D , E qRT-PCR was conducted to assess the differences in mRNA expression of the above 16 genes after RBM15 knockdown in H1975 and HCC827 cells. T test. F , G Western blot assessment of the correlation between RBM15 and SPOCK1 protein expression. H , I RIP-PCR validated RBM15 and m6A binding to SPOCK1 mRNA. T test. J Changes in total RNA m6A modification levels after 48-h DAA treatment. T test. K Alteration of SPOCK1 mRNA levels in H1975 cells and HCC827 cells after DAA treatment. T test. L , M Western blot and qRT-PCR demonstrated reduced SPOCK1 expression in H1975-OR cells and PDO1-OR. T test. N Osimertinib IC 50 value of H1975-OR cells transfected with si-NC, si-RBM15, or si-SPOCK1. T test. O Osimertinib IC 50 value of PDO1-OR transfected with sh-NC, sh-RBM15, or sh-SPOCK1. T test. P , Q Effects of RBM15 and SPOCK1 knockdown on H1975-OR and PDO1-OR apoptosis assessed using flow cytometry and TUNEL staining. T test. Error bars represent the means ± SDs. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: When the tumors reached a longitudinal diameter of 5 mm, the mice were randomly assigned to each group and subsequently treated orally with an equal volume of osimertinib mesylate (Selleck, S5078, TX, 20 mg/kg/day) or corn oil by gavage every 2 days.

Techniques: Generated, RNA Sequencing, Modification, Quantitative RT-PCR, Expressing, Knockdown, Western Blot, Binding Assay, Transfection, Flow Cytometry, TUNEL Assay, Staining

Journal: Oncogene

Article Title: RBM15 facilitates osimertinib resistance of lung adenocarcinoma through m6A-dependent epigenetic silencing of SPOCK1

doi: 10.1038/s41388-024-03220-z

Figure Lengend Snippet: A , B GSEA pathway enrichment was performed using DEGs. C Volcano plot of RNA-seq results displaying DEGs in H1975 and H1975-OR. D , E Western blot and immunofluorescence detection of protein expression of EMT-related indicators (scale bar: 50 μm). T test. F Immunofluorescence detection of protein expression of EMT-related indicators in PDO-OS and PDO1-OR (scale bar: 25 μm). T test. G Change in CDH1, ZEB1, AXL, and RBM15 expression in osimertinib refractory tissues and matched pre-treatment tissues. H – J Spearman’s correlation of change in CDH1, ZEB1, AXL, and RBM15 expression in eight patients with matched pre- and post-osimertinib treatment. Pearson test. K , L Western blot and immunofluorescence assays reveal changes in protein expression of EMT indicators in H1975-OR and PDO1-OR after RBM15 knockdown. M Immunofluorescence assays reveal changes in protein expression of EMT indicators in PDO1-OR after RBM15 and SPOCK1 knockdown (scale bar: 50 μm). T test. Error bars represent the means ± SDs. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: When the tumors reached a longitudinal diameter of 5 mm, the mice were randomly assigned to each group and subsequently treated orally with an equal volume of osimertinib mesylate (Selleck, S5078, TX, 20 mg/kg/day) or corn oil by gavage every 2 days.

Techniques: RNA Sequencing, Western Blot, Immunofluorescence, Expressing, Knockdown

Journal: Oncogene

Article Title: RBM15 facilitates osimertinib resistance of lung adenocarcinoma through m6A-dependent epigenetic silencing of SPOCK1

doi: 10.1038/s41388-024-03220-z

Figure Lengend Snippet: A H1975 cells were exposed to 1 μm osimertinib and cultured for 20 days to select H1975-DTP cells (top panel). H1975 and H1975-DTP cells were incubated with osimertinib for 48 h, cell viability was measured using the CCK-8 assay, and the IC 50 was calculated (bottom panel). T test. B Western blot investigation of EGFR, ERK, and AKT protein levels and their phosphorylation. C Western blot assessment of RBM15 protein expression levels in H1975 and H1975-DTPCs. D , E Western blot and immunofluorescence assessment of EMT marker proteins in H1975 and H1975-DTP cells (scale bar: 50 μm). T test. F A model diagram demonstrating that RBM15 suppresses SPOCK1 mRNA expression through m6A modification, enhancing EMT-mediated osimertinib resistance in refractory tumors. This figure was created using the BioRender website ( https://biorender.com ). Error bars represent the means ± SDs. *** P < 0.001, **** P < 0.0001.

Article Snippet: When the tumors reached a longitudinal diameter of 5 mm, the mice were randomly assigned to each group and subsequently treated orally with an equal volume of osimertinib mesylate (Selleck, S5078, TX, 20 mg/kg/day) or corn oil by gavage every 2 days.

Techniques: Cell Culture, Incubation, CCK-8 Assay, Western Blot, Phospho-proteomics, Expressing, Immunofluorescence, Marker, Modification

Journal: Cancer research

Article Title: EGFR Activates a TAZ-driven Oncogenic Program in Glioblastoma

doi: 10.1158/0008-5472.CAN-20-2773

Figure Lengend Snippet: KEY RESOURCES

Article Snippet: Osimertinib (0.5μM for screening) , Medkoo , # 206042.

Techniques: Virus, Recombinant, Enzyme-linked Immunosorbent Assay, Sample Prep, Plasmid Preparation, shRNA, Control, Software, Negative Control

Journal: F1000Research

Article Title: Ex-vivo drug screening of surgically resected glioma stem cells to replace murine avatars and provide personalise cancer therapy for glioblastoma patients

doi: 10.12688/f1000research.135809.1

Figure Lengend Snippet: Drug compounds and concentrations used on 384 well drug plate.

Article Snippet: AZD9291 (Osimertinib) , Stratech (Selleckchem) , 5 , 2.5 , 1.25 , 0.625.

Techniques: Concentration Assay