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: Journal of cellular physiology

Article Title: Blocking Fibrotic Signaling In Fibroblasts from Patients with Carpal Tunnel Syndrome

doi: 10.1002/jcp.25901

Figure Lengend Snippet: Gene expression analysis using qRT-PCR of known CTS fibrosis marker genes Col1, 3, CTGF, SERPINE1 upon cytokine receptor level inhibition. CTS fibroblasts were treated with TGF-β1 and TβRI (SD208), PDGFR (AG1296), EGFR (Lapatinib) and VEGFR (Axitinib) inhibitor for 24 hours. Gene expression is normalized to vehicle control. All data are expressed as mean ± SE. n=5. (* indicates p < 0.05, # indicates p < 0.01).

Article Snippet: Lapatinib (IC50: 10.8 nM) (Selleck Chemical, Houston, TX, S1028) is a small molecule kinase inhibitor that targets both EGFR1 and 2 (Her2) ( Moy et al., 2007 ).

Techniques: Gene Expression, Quantitative RT-PCR, Marker, Inhibition, Control

Journal: Journal of cellular physiology

Article Title: Blocking Fibrotic Signaling In Fibroblasts from Patients with Carpal Tunnel Syndrome

doi: 10.1002/jcp.25901

Figure Lengend Snippet: Smad reporter activity comparing cytokine receptor inhibition. Comparison of CTS fibroblasts treated with TGF-β1 and TβRI (SD208), PDGFR (AG1296), EGFR (Lapatinib) and VEGFR (Axitinib) inhibitor for 24 hours. Data are expressed as relative fold change from vehicle control. All data are expressed as mean ± SE. n=3. (# indicates p < 0.01).

Article Snippet: Lapatinib (IC50: 10.8 nM) (Selleck Chemical, Houston, TX, S1028) is a small molecule kinase inhibitor that targets both EGFR1 and 2 (Her2) ( Moy et al., 2007 ).

Techniques: Activity Assay, Inhibition, Comparison, Control

Journal: Scientific Reports

Article Title: Mesenchymal Stem Cells Exploit Extracellular Matrix as Mechanotransducer

doi: 10.1038/srep02425

Figure Lengend Snippet: (a) hMSCs cultured on single Fn fibers in osteogenic induction medium for 7 days with or without constant exposure to function-blocking antibodies. Percentage of ALP positive hMSCs is shown in the presence of function-blocking antibodies against integrin α5β1 (red), integrin αvβ3 (blue) or without antibodies (yellow). (b) Percentage of ALP positive hMSCs when cultured for 7 days on single Fn fibers in the presence (green) or absence (yellow) of the EGFR inhibitor GW572016 in osteogenic induction medium is shown. Data shown in a and b represent mean ± s.d. (n = 5). Asterisk p < 0.05 versus no antibody treatment (P = 0.0385 for α5β1 in (a)). Two asterisks: p < 0.01 versus no antibody treatment (P = 0.0038 for αvβ3 in (a)) or no drug treatment (p = 0.0092 for (b)).

Article Snippet: To block the activity of ErbB receptors, hMSCs (50 × 10 3 cells/ml) were allowed to attach for 1 hour on single Fn fibers in growth medium supplemented with GW572016 (1 μM, Lapatinib, Axon Medchem), and were subsequently cultured for 7 days in osteogenic induction medium supplemented with GW572016.

Techniques: Cell Culture, Blocking Assay

Journal: Disease Markers

Article Title: Predicting the Efficacy of HER2-Targeted Therapies: A Look at the Host

doi: 10.1155/2017/7849108

Figure Lengend Snippet: Anti-HER2 therapies and their immunostimulatory properties. The monoclonal antibodies trastuzumab and pertuzumab, in addition to inhibiting intracellular signaling downstream of HER2 activation (i.e., homo/heterodimerization and proteolytic cleavage of the HER2 extracellular domain), induce an antitumor immune response in the tumor microenvironment. Trastuzumab and pertuzumab bind to the extracellular domain of HER2 and, through their Fc portions, engage antibody-dependent cell-mediated cytotoxicity (ADCC) and phagocytosis (ADCP) in Fc receptor-positive innate immune cells (i.e., NK lymphocytes, macrophages, monocytes, and neutrophils). Immune complex and opsonized tumor fragments are recognized and taken up by dendritic cells via the Fc receptor. Dendritic cells and other antigen-presenting cells (e.g., macrophages) present tumor antigens through MHC-II molecules to CD4+ T-helper lymphocytes, which release interferon- γ (IFN γ ), interleukin 2 (IL2), and IL21 to enhance the cytotoxic T cell response. Antigens presented by MHC-I molecules directly stimulate CD8+ cytotoxic T lymphocytes. CD8+ T cells can also recognize tumor antigens presented on MHC-I molecules by cancer cells themselves and initiate a cytotoxic response. Tyrosine kinase inhibitors (TKIs) (lapatinib and neratinib) block the kinase domain activity of HERs, disrupting the oncogenic signals that lead to proliferation, migration, invasion, and survival of cancer cells. In contrast to neratinib, lapatinib, in addition to blocking the TK domain of HER2 and HER1, affects the accumulation of HER2 on the surface of BC cells, leading to an increase in ADCC when combined with trastuzumab.

Article Snippet: Lapatinib (GW572016, Tyverb/Tykerb; Novartis, Switzerland) reversibly inhibits the intracellular tyrosine kinase domains of EGFR and HER2, inducing cell cycle arrest and apoptosis in BC cell lines through the cleavage of PARP and the activation of caspase 3 [ ].

Techniques: Bioprocessing, Activation Assay, Blocking Assay, Activity Assay, Migration

Journal: Disease Markers

Article Title: Predicting the Efficacy of HER2-Targeted Therapies: A Look at the Host

doi: 10.1155/2017/7849108

Figure Lengend Snippet: Evaluation of HER2-addiction and immune biomarkers in randomized trials investigating anti-HER2 targeted therapies.

Article Snippet: Lapatinib (GW572016, Tyverb/Tykerb; Novartis, Switzerland) reversibly inhibits the intracellular tyrosine kinase domains of EGFR and HER2, inducing cell cycle arrest and apoptosis in BC cell lines through the cleavage of PARP and the activation of caspase 3 [ ].

Techniques: Polymerase Chain Reaction, Adjuvant