Journal: eBioMedicine

Article Title: Single-nucleus RNA sequencing reveals heterogenous microenvironments and specific drug response between cervical squamous cell carcinoma and adenocarcinoma

doi: 10.1016/j.ebiom.2023.104846

Figure Lengend Snippet: The transcriptomic heterogeneities of malignant cells in CSCC and CAde. (a – b) t-SNE plots of malign epithelial cells, colored by cell clusters and individual samples, respectively. (c) Dot plots revealed the gene expression profiles of each malignant cell cluster. (d) Heatmap revealed the molecular functions of each malignant cell cluster based on GSVA analysis (MSigDB_C6). (e) The box plot depicted the stemness score of malignant cells in CSCC and CAde. The P -value was calculated by Wilcoxon Rank Sum Test. (f) Forest plot of the identified differentially expressed genes from snRNA-seq data for CSCC and CAde regarding overall survival. (g) Representative IHC staining of MTSS1 in collected CSCC (n = 32) and CAde (n = 11) samples. ∗∗∗ P < 0.001. The P -value was calculated by Wilcoxon Rank Sum Test. (h) Representative IHC staining of POLA1 in collected CSCC (n = 32) and CAde (n = 11) samples. ∗∗∗ P < 0.001. The P -value was calculated by Wilcoxon Rank Sum Test. (i) The ROC curve (sensitivity against 1-specificity) of identified specific gene signature for classifying CSCC and CAde based on TCGA CESC cohort. (j) Dot plots displayed the abnormally activated transcription factors (TFs) for CSCC and CAde. (k) Radar plot displayed the expression profiles of identified TFs in CC cell lines. The distance from the dots to the center of the circle represented the log2 (relative gene expression). (l) Gene regulatory network among TFs and target genes in malignant epithelial cells. Significant genes meant the adjusted P -value <0.01 and |Log2FC| > 0.25. P -value was calculated by Wilcoxon Rank Sum t-test. (m) Scatter plots of switch point of a certain drugs in CSCC and CAde. The switch point of certain drug less than 0.1 was defined as sensitivity, and the sensitivity of drugs in CSCC and CAde were denoted by color. The Switch Point value of 0 was converted to 0.009 for plotting. (n) IC50 values of selected drugs (Pyrimethamine, Lapatinib, Dasatinib and Doramapimod) in CaSki and HeLa cell line. Each drug had at least three biological replicates in each cell line. (o) Morphological changes of constructed organoids from CSCC and CAde samples after being treated with Lapatinib and Dasatinib. (p) IC50 values of selected drugs (Pyrimethamine, Lapatinib, Dasatinib and Doramapimod) in CSCC- and CAde organoids. Each drug had at least three biological replicates in each constructed organoid.

Article Snippet: The details of drugs were as follows: Pyrimethamine (MCE, #HY-18062), Lapatinib (MCE, #HY-50898), Dasatinib (MCE, #HY-10181), and Doramapimod (MCE, #HY-10320).

Techniques: Gene Expression, Immunohistochemistry, Expressing, Construct

Journal: Journal of clinical medicine

Article Title: Liposomal Lapatinib in Combination with Low-Dose Photodynamic Therapy for the Treatment of Glioma.

doi: 10.3390/jcm8122214

Figure Lengend Snippet: Figure 1. (A) The encapsulation efficiency of lapatinib within the liposomal formulation prepared with up to 2000 nmol lapatinib addition and the corresponding liposome size of the resultant liposomes. (B) Long-term stability of the liposomal lapatinib formulation prepared with 500 nmol lapatinib addition monitored by both z-average diameter and the respective polydispersity indices. (mean ± S.D., n = 3).

Article Snippet: A chloroform suspension of lapatinib ditosylate (925.46 g/mol, 10 mg/mL; Selleckchem, Radnor, PA, USA) was prepared, and 500 nmol of lapatinib ditosylate equivalent (46.3 μL) was added to the lipid mixture.

Techniques: Encapsulation, Formulation, Liposomes

Journal: Journal of clinical medicine

Article Title: Liposomal Lapatinib in Combination with Low-Dose Photodynamic Therapy for the Treatment of Glioma.

doi: 10.3390/jcm8122214

Figure Lengend Snippet: Figure 2. Live-cell imaging-based of protoporphyrin IX (PpIX) fluorescence within mitochondria and cytoplasm and the fraction between the two cellular locations as well as LD50 data when ALA and liposome-encapsulated lapatinib treatments were combined. (A) Percentage changes in PpIX production in the mitochondria following control (+HBEGF 5 ng/mL an epidermal growth factor receptor (EGFR) ligand commonly found in the brain for activation) and liposome-encapsulated lapatinib (500 nM) (+HBEGF) (* p < 0.05) (n = 3 biological replicates). (B) The fraction of PpIX in the mitochondria compared to the cytoplasm with and without lapatinib for all cell lines (* p < 0.05) (n = 3 biological replicates. (C) LD50 values of four glioma cell lines and one purported glioma cancer stem cell line (GS2) of PDT alone and PDT + Lapatinib, note LD50 of ALA, was chosen as a surrogate for PDT dose in these studies. A significant decrease in LD50 for the two EGFR positive cell lines U87 and U87vIII was defined by p < 0.05 (n = 3).

Article Snippet: A chloroform suspension of lapatinib ditosylate (925.46 g/mol, 10 mg/mL; Selleckchem, Radnor, PA, USA) was prepared, and 500 nmol of lapatinib ditosylate equivalent (46.3 μL) was added to the lipid mixture.

Techniques: Live Cell Imaging, Control, Activation Assay

Journal: Journal of clinical medicine

Article Title: Liposomal Lapatinib in Combination with Low-Dose Photodynamic Therapy for the Treatment of Glioma.

doi: 10.3390/jcm8122214

Figure Lengend Snippet: Figure 3. (A) MR images of central slices of five different U87 tumours prior to assignment to treatment groups including (a–e) contrast-enhanced T1w; and (f–j) T2w images (B) H&E staining of tumours at endpoint for each treatment groups. (C) Survival post-treatment of animals in each cohort (solid black line—control; solid grey line—lapatinib; short, broken line—photodynamic therapy (PDT) alone; broken line—PDT + lapatinib.

Article Snippet: A chloroform suspension of lapatinib ditosylate (925.46 g/mol, 10 mg/mL; Selleckchem, Radnor, PA, USA) was prepared, and 500 nmol of lapatinib ditosylate equivalent (46.3 μL) was added to the lipid mixture.

Techniques: Staining, Control

Journal: Advanced Science

Article Title: Semaphorin 3E–Plexin D1 Axis Drives Lung Fibrosis through ErbB2‐Mediated Fibroblast Activation

doi: 10.1002/advs.202415007

Figure Lengend Snippet: P61‐Sema3E produces a pro‐fibrotic effect through the activation of ErbB2. A) Western blot analysis of the levels of ErbB2, P‐ErbB2, AKT, P‐AKT, ERK1/2, and P‐ERK1/2 in primary human lung fibroblasts (PHLFs) after stimulation with different concentrations of P61‐Sema3E for 2 h. B) Western blot analysis of ErbB2, P‐ErbB2, AKT, P‐AKT, ERK1/2, and P‐ERK1/2 expression in PHLFs treated with PlexinD1 siRNA or Scrambled siRNA following P61‐Sema3E induction. C) Co‐immunoprecipitation analysis of Plexin D1 and ErbB2, FLAG‐tagged Plexin D1 plasmid were transfected into PHLFs. D) Co‐immunoprecipitation analysis of Plexin D1 and ErbB2, PHLFs were treated with P61‐Sema3E for 2 h. E) Western blot analysis of Fibronectin, Col1a1, and α‐SMA expression in PHLFs treated with the ErbB2 inhibitor Lapatinib or PBS following P61‐Sema3E stimulation for 48 h. Data are represented as the mean ± SEM of three independent experiments. Statistical analyses were performed using one‐way ANOVA test. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The ErbB2 inhibitor Lapatinib, was obtained from MedChemExpress (CM00323).

Techniques: Activation Assay, Western Blot, Expressing, Immunoprecipitation, Plasmid Preparation, Transfection

Journal: Advanced Science

Article Title: Semaphorin 3E–Plexin D1 Axis Drives Lung Fibrosis through ErbB2‐Mediated Fibroblast Activation

doi: 10.1002/advs.202415007

Figure Lengend Snippet: Downregulation of Sema3E attenuates BLM‐induced pulmonary fibrosis. A,B) Histological analysis of lung fibrosis severity in mice following BLM induction. (A) Representative images of lung sections stained with H&E, Masson's trichrome, and Sirius red to assess fibrosis severity. (B) Bar graph showing the quantitative mean score of fibrosis severity. Samples include AAV9‐NC saline mice ( n = 5), AAV9‐ShSema3E saline mice ( n = 5), AAV9‐NC BLM mice ( n = 5), and AAV9‐ShSema3E BLM mice ( n = 5). C) Quantification of hydroxyproline contents in AAV9‐NC mice and AAV9‐ShSema3E mice after BLM challenge. D,E) Western blot and RT‐qPCR analysis of Fibronectin, Col1a1, and α‐SMA protein and mRNA expression in lung homogenates from the mentioned mouse groups. F) Western blot analysis of ErbB2, P‐ErbB2, AKT, P‐AKT, ERK1/2, and P‐ERK1/2 expression in lung homogenates from the mentioned mouse groups. Samples include AAV9‐NC saline mice ( n = 3), AAV9‐ShSema3E saline mice ( n = 3), AAV9‐NC BLM mice ( n = 3), and AAV9‐ShSema3E BLM mice ( n = 3). Data are represented as the mean ± SEM. Statistical analyses were performed using one‐way ANOVA tests. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The ErbB2 inhibitor Lapatinib, was obtained from MedChemExpress (CM00323).

Techniques: Staining, Saline, Western Blot, Quantitative RT-PCR, Expressing

Journal: Advanced Science

Article Title: Semaphorin 3E–Plexin D1 Axis Drives Lung Fibrosis through ErbB2‐Mediated Fibroblast Activation

doi: 10.1002/advs.202415007

Figure Lengend Snippet: Sema3E deficiency in fibroblasts protects mice from BLM‐induced lung injury and fibrosis. A,B) Representative lung sections from Sema3E‐C and Sema3E‐CKO mice treated with saline or BLM, stained with H&E, Masson's Trichrome, and Sirius Red (Panel A, left). Panel B (right) shows quantitative fibrosis scores. Each group consisted of five mice ( n = 5). C) Quantification of hydroxyproline contents in Sema3E‐CKO and Sema3E‐C mice after BLM challenge. D,E) Western blot and RT‐qPCR analyses of Fibronectin, Col1a1, and α‐SMA protein and mRNA levels in lung homogenates from each group ( n = 5). F) Western blot analysis of ErbB2, P‐ErbB2, AKT, P‐AKT, ERK1/2, and P‐ERK1/2 in lung homogenates from the same groups of three mice( n = 3). Data are expressed as the mean ± SEM. Statistical significance was determined by one‐way ANOVA (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: The ErbB2 inhibitor Lapatinib, was obtained from MedChemExpress (CM00323).

Techniques: Saline, Staining, Western Blot, Quantitative RT-PCR

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Patient-derived xenografts and matched cell lines identify pharmacogenomic vulnerabilities in colorectal cancer

doi: 10.1158/1078-0432.CCR-18-3440

Figure Lengend Snippet: A, Heat map showing the relative expression (log2 ratio FPKM) of receptor tyrosine kinases (RTKs) as identified by RNA-seq analysis of 29 XLs. RTKs listed here (n=32) represent the only members of this class of protein passing the filtration process during RNA-seq analysis. KRAS, BRAF and NRAS mutational status is also indicated. B, Scatter plot displaying the correlation between absolute ERBB2 expression (FPKM, x-axis) and relative ERBB2 copy-number variation (CNV, log2ratio) in the 29 XLs, as assessed by RNA-seq and WES analyses, respectively. ERBB2-overexpressing and amplified XLs (CRC0080-XL and CRC01086-XL) are indicated. C, Short-term viability assay showing the effects of single dose of trastuzumab (TMAB, 25 μg/mL), lapatinib (LAP, 120 nM), and combination of lapatinib and trastuzumab (LAP+TMAB) on ERBB2-amplified CRC0186-XL (left panel) and CRC0080-XL (right panel). Viability was assessed by measuring ATP content after 6 days of treatment. Data represent the values obtained from at least three independent experiments in triplicate, normalized on control DMSO-treated cells; error bars indicate SD (lapatinib vs lapatinib+trastuzumab in CRC0186-XL: ***, P = 0.001; lapatinib vs lapatinib+trastuzumab: **, P = 0.0036 in CRC0080-XL; unpaired Student’s t test). IC50 values were calculated from dose inhibition curves using GraphPad Prism software. D, Activation status/phosphorylation of HER2 receptor and downstream pathway in dose-response experiments in CRC0186-XL. Cells were treated with the indicated concentrations of trastuzumab (TMAB), lapatinib (Lap), or the combination of both drugs for 4 hours. Cell extracts were immunoblotted with the indicated antibodies. HSP90 was used as loading control. E, In vivo tumor growth curves of CRC0186-PDX (n=6) and CRC0080-PDX (n=6) treated with the indicated modalities. Data represent the mean of tumor volume at each time point; error bars indicate S.E.M. (lapatinib vs lapatinib+trastuzumab: ***, P < 0.001 by two-way Anova Test). Response curves for both models were previously reported in (49) and are reshown here for comparative purposes.

Article Snippet: Established tumors (average volume 400 mm 3 ) were treated with the following regimens, either single-agent or in combination: lapatinib (Carbosynth) 100 mg/kg daily (vehicle, 0.5% methylcellulose, 0.2% Tween-80); trastuzumab (Roche) 30 mg/kg once weekly (vehicle: physiological saline).

Techniques: Expressing, RNA Sequencing, Filtration, Amplification, Viability Assay, Control, Inhibition, Software, Activation Assay, Phospho-proteomics, In Vivo

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Patient-derived xenografts and matched cell lines identify pharmacogenomic vulnerabilities in colorectal cancer

doi: 10.1158/1078-0432.CCR-18-3440

Figure Lengend Snippet: A, Long-term viability assay showing the effect of different indicated doses of lapatinib in absence or in presence of doses of 25 μg/ml of trastuzumab on ERBB2-amplified CRC0186-XL transduced with lentiviral vector overexpressing BRAF V600E, KRAS G13D or PIK3CA H1047R mutants. Cells transduced with empty lentiviral vector were used as control (CTRL). Cell viability was assessed by crystal violet staining after 15 days of treatment. Representative wells were photographed and reported. B, Activation status/phosphorylation of HER2 receptor and downstream pathway in CRC0186-XL transduced with lentiviral vector overexpressing the indicated alleles. Cells transduced with empty lentiviral vector were used as control. Cells were treated with the combination of 500 nM of lapatinib and 10 μg/ml of trastuzumab (L+T) for 4 hours. Cell extracts were immunoblotted with the indicated antibodies. Vinculin was used as loading control.

Article Snippet: Established tumors (average volume 400 mm 3 ) were treated with the following regimens, either single-agent or in combination: lapatinib (Carbosynth) 100 mg/kg daily (vehicle, 0.5% methylcellulose, 0.2% Tween-80); trastuzumab (Roche) 30 mg/kg once weekly (vehicle: physiological saline).

Techniques: Viability Assay, Amplification, Transduction, Plasmid Preparation, Control, Staining, Activation Assay, Phospho-proteomics

Journal: Oncotarget

Article Title: TP53 mutated glioblastoma stem-like cell cultures are sensitive to dual mTORC1/2 inhibition while resistance in TP53 wild type cultures can be overcome by combined inhibition of mTORC1/2 and Bcl-2

doi: 10.18632/oncotarget.11205

Figure Lengend Snippet: Significant associations between mutated genes and drug response

Article Snippet: The following small molecule compounds targeting the RTK/Ras/PI3K pathway were diluted in DMSO (Sigma-Aldrich, St. Louis, USA): lapatinib ditosylate was obtained from Santa Cruz Biotechnology (Dallas, USA) (catalog number 202205A).

Techniques: Mutagenesis