Journal: Toxicological Sciences

Article Title: Selective inhibition of integrin αvβ6 leads to rapid induction of urinary bladder tumors in cynomolgus macaques

doi: 10.1093/toxsci/kfac128

Figure Lengend Snippet: Inhibition of αvβ6 integrin or TGFβR1 induce proliferation in primary human bladder epithelial cells in culture. All proliferation measurements were assessed following 3 days of exposure to treatments. Selective αvβ6 integrin inhibitor, MORF-627 (A). Structurally distinct tool αvβ6 integrin inhibitor (B). TGFβR1 multikinase inhibitor (C). Proliferative changes induced by MORF-627 were reversed by concomitant incubation of test agent with exogenous TGF-β (D). Graphs represent 3 independent experiments, with each concentration run in triplicate per study. All data are plotted as raw luminescence mean ± SEM; * p < .05, ** p < .01, *** p < .001, and **** p < .0001 indicate statistical significance compared with bleomycin IgG isotype group, using 2-tailed unpaired t -test with Welsh’s correction.

Article Snippet: An anti-αvβ6 antibody ( Weinreb et al. , 2004 ) without effector function was generated by Morphic Therapeutic at ATUM Bio, based on the sequence of the variable fragment (Fab) of 3G9 (US patent 8,153,126 B2) fused with the constant fragment (Fc) of mouse IgG1 backbone.

Techniques: Inhibition, Incubation, Concentration Assay

Journal: Toxicological Sciences

Article Title: Selective inhibition of integrin αvβ6 leads to rapid induction of urinary bladder tumors in cynomolgus macaques

doi: 10.1093/toxsci/kfac128

Figure Lengend Snippet: Inhibition of αvβ6 integrin or TGFβR1 induce proliferation in primary human alveolar epithelial cells in culture. All proliferation measurements were assessed following 3 days of exposure to treatments. Selective αvβ6 integrin inhibitor, MORF-627 (A). Structurally distinct tool αvβ6 integrin inhibitor (B). TGFβR1 multikinase inhibitor (C). Proliferative changes induced by MORF-627 were reversed by concomitant incubation of test agent with exogenous TGF-β (D). Graphs represent 3 independent experiments, with each concentration run in triplicate per study. All data are plotted as raw luminescence mean ± SEM; * p < .05, ** p < .01, *** p < .001, and **** p < .0001 indicate statistical significance compared with bleomycin IgG isotype group, using 2-tailed unpaired t -test with Welsh’s correction.

Article Snippet: An anti-αvβ6 antibody ( Weinreb et al. , 2004 ) without effector function was generated by Morphic Therapeutic at ATUM Bio, based on the sequence of the variable fragment (Fab) of 3G9 (US patent 8,153,126 B2) fused with the constant fragment (Fc) of mouse IgG1 backbone.

Techniques: Inhibition, Incubation, Concentration Assay

Journal: Toxicological Sciences

Article Title: Selective inhibition of integrin αvβ6 leads to rapid induction of urinary bladder tumors in cynomolgus macaques

doi: 10.1093/toxsci/kfac128

Figure Lengend Snippet: Tool αvβ6 inhibitor and TGFβR1 selective inhibitor increased biliary epithelial cell proliferation in DDC-induced biliary fibrosis model. A, Representative images of IHC staining for CK19 expressing epithelial cells in healthy mice ( n = 5) and mice fed with DDC diet with or without treatment ( n = 9–15/group). Scale bar = 200 µm. B, IHC quantification of CK19 expressing biliary epithelial cells. (**** p < .0001 indicate statistical significance compared with DDC vehicle group, using 1-way ANOVA with Dunnett’s post hoc test.) C, IHC quantification of Ki67 positive proliferating cells (* p < .05, ** p < .01 indicate statistical significance compared with DDC vehicle group using 2-tailed Student’s T test). All data are shown as means ± SEM.

Article Snippet: An anti-αvβ6 antibody ( Weinreb et al. , 2004 ) without effector function was generated by Morphic Therapeutic at ATUM Bio, based on the sequence of the variable fragment (Fab) of 3G9 (US patent 8,153,126 B2) fused with the constant fragment (Fc) of mouse IgG1 backbone.

Techniques: Immunohistochemistry, Expressing

Journal: Toxicological Sciences

Article Title: Selective inhibition of integrin αvβ6 leads to rapid induction of urinary bladder tumors in cynomolgus macaques

doi: 10.1093/toxsci/kfac128

Figure Lengend Snippet: Anti-αvβ6 antibody (3G9) increased biliary epithelial cell proliferation in DDC-induced biliary fibrosis model. A, IHC quantification of CK19 expressing epithelial cells in healthy mice and mice fed with DDC diet with or without treatment ( n = 10/group). B, IHC quantification of Ki67 positive proliferating cells. C, KEGG pathway analysis of bulk RNA sequencing data from liver tissues. All data are shown as means ± SEM. * p < .05, ** p < .01, *** p < .001, and **** p < .0001 indicate statistical significance compared with DDC vehicle group, using 1-way ANOVA with Dunnett’s post hoc test.

Article Snippet: An anti-αvβ6 antibody ( Weinreb et al. , 2004 ) without effector function was generated by Morphic Therapeutic at ATUM Bio, based on the sequence of the variable fragment (Fab) of 3G9 (US patent 8,153,126 B2) fused with the constant fragment (Fc) of mouse IgG1 backbone.

Techniques: Expressing, RNA Sequencing

Journal: Toxicological Sciences

Article Title: Selective inhibition of integrin αvβ6 leads to rapid induction of urinary bladder tumors in cynomolgus macaques

doi: 10.1093/toxsci/kfac128

Figure Lengend Snippet: Anti-αvβ6 antibody (3G9) promoted cell proliferation in bleomycin-induced lung fibrosis model. A, KEGG pathway analysis of bulk RNA sequencing data from lung tissues. mRNA levels of cell cycle genes, (B) Mki67 and (C) Pcna, in lung tissues. All data are shown as means ± SEM. * p < .05, ** p < .01, *** p < .001, and **** p < .0001 indicate statistical significance compared with bleomycin IgG isotype group, using 1-way ANOVA with Dunnett’s post hoc test.

Article Snippet: An anti-αvβ6 antibody ( Weinreb et al. , 2004 ) without effector function was generated by Morphic Therapeutic at ATUM Bio, based on the sequence of the variable fragment (Fab) of 3G9 (US patent 8,153,126 B2) fused with the constant fragment (Fc) of mouse IgG1 backbone.

Techniques: RNA Sequencing

Journal: Frontiers in Pharmacology

Article Title: Integrin α4β1/VCAM-1 Interaction Evokes Dynamic Cell Aggregation Between Immune Cells and Human Lung Microvascular Endothelial Cells at Infectious Hemolysis

doi: 10.3389/fphar.2021.653143

Figure Lengend Snippet: (A) Images of RBCs/mo-DCs aggregates captured by Keyence fluorescence microscope. RBCs and mo-DCs were pre-labeled with WGA-555 and Hoechst dye, respectively. Top panels show medium control; middle panels show LPS treated RBCs group (LPS 50 μg/ml for 24 h); bottom panels show the RBCs/mo-DCs group with RBC pretreated with 10 µM of the cyclic peptide inhibitor of integrin α4β1. White arrows pointed RBCs/mo-DCs aggregates. (B) The imaging strategy to distinguish cell aggregation with the corresponding three cell types in cell suspension. RBCs, mo-DCs, and HLMVEC were pre-labeled with WGA-555, Hoechst dye and WGA-488 respectively. Accordingly, aggregates were defined as RBCs/mo-DCs (white arrows), RBCs/HLMEC (yellow arrows), and RBCs/mo-DCs/HLMEC (pink arrows). (C) Quantification of cell aggregation calculated by imaging analysis software (left two panels). Several controls were used to access the ability of cell aggregation as indicated. Flow cytometric analysis of the integrin β1 expression on RBCs (middle panel). Integrin β1 on RBCs treated by LPS increased significantly compared to untreated control (graph bar panel, right side of flow cytometry picture). The α4β1 peptide inhibitor (10 µM) or Natalizmab (10 μg/ml) blocked cell-cell aggregation of RBCs/mo-DCs in a concentration-dependent manner while peptide control or Methylprednisolone alone had no effect on cell aggregation (Bottom right panel). All data were representative of mean ± SD, with at least three biological replicates. * p < 0.05, *** p < 0.001.

Article Snippet: The cyclic peptide inhibitor of integrin α4β1 (azide modified peptide: MePhe-Leu-Asp-Val-Aib-Lys) ( ) control peptides (cyclic MePhe-Ala-Ala-Ala-Aib-Lys), and peptide drug conjugates (PDC), cyclic peptide inhibitor of integrin α4β1 conjugated methylprednisolone (α4β1/Methrol), and control peptide/methrol were synthesized using solid-supported chemistry by InnoPep Inc (San Diego, CA, United States).

Techniques: Fluorescence, Microscopy, Labeling, Control, Imaging, Suspension, Software, Expressing, Flow Cytometry, Concentration Assay

Journal: Frontiers in Pharmacology

Article Title: Integrin α4β1/VCAM-1 Interaction Evokes Dynamic Cell Aggregation Between Immune Cells and Human Lung Microvascular Endothelial Cells at Infectious Hemolysis

doi: 10.3389/fphar.2021.653143

Figure Lengend Snippet: (A) LPS pretreated RBCs induced cell-cell aggregation on RBCs/mo-DCs, RBCs/HLMVEC, and RBCs/mo-DCS/HLMVEC (* p < 0.05 vs. untreated groups). Such effects were blocked by α4β1/Methrol (# p < 0.05). (B) LPS or L-sup enhanced mo-DCs migration (* p < 0.05, ## p < 0.01 vs. C-sup; * p < 0.05, L-sup vs. LPS alone) which was attenuated by α4β1/Methrol (* p < 0.01 vs. L-sup). (C) α4β1/Methrol decreased L-sup induced mo-DCs migration in a concentration-dependent manner [* p < 0.05 vs. peptide drug conjugate (PDC control)]. Data were representative of mean ± SD of three experiments. (D) GILZ-expressing mo-DCs analyzed by flow cytometry (dark color: PDC control; light black color: 10 µM α1β4/Methrol treatment for 24 h). (E) MFI values of GILZ-expressing mo-DCs quantitated by flow cytometry. α4β1/Methrol significantly increased the number of mo-DCs expressing GILZ as compared to PDC control (* p < 0.05).

Article Snippet: The cyclic peptide inhibitor of integrin α4β1 (azide modified peptide: MePhe-Leu-Asp-Val-Aib-Lys) ( ) control peptides (cyclic MePhe-Ala-Ala-Ala-Aib-Lys), and peptide drug conjugates (PDC), cyclic peptide inhibitor of integrin α4β1 conjugated methylprednisolone (α4β1/Methrol), and control peptide/methrol were synthesized using solid-supported chemistry by InnoPep Inc (San Diego, CA, United States).

Techniques: Migration, Concentration Assay, Control, Expressing, Flow Cytometry

Journal: Frontiers in Pharmacology

Article Title: Integrin α4β1/VCAM-1 Interaction Evokes Dynamic Cell Aggregation Between Immune Cells and Human Lung Microvascular Endothelial Cells at Infectious Hemolysis

doi: 10.3389/fphar.2021.653143

Figure Lengend Snippet: Schematic diagram of the α4β1/Methrol reversing dendritic cell dysfunction in LPS-induced hemolysis. LPS-induced hemolysis resulted in an increase of integrin α4β1 expression on RBCs that subsequently mediated cell-cell aggregation, released cytokines/chemokines and evoked immune cell responses. LPS increased integrin α4β1 expression to facilitate cell-cell interaction of RBCs/mo-DCs, RBCs/HLMVEC, and RBCs/mo-DCs/HLMVEC. Cell aggregation of RBCs and mo-DCs further enhanced cytokines release and activation of NF-kB pathway in immune T cells. Chemokines/cytokines induced mo-DCs migration to HLMVEC. α4β1/VCAM-1 is an interconnective molecule for RBCs, mo-DCs, and HLMVEC aggregation at inflamed site. α4β1/Methrol blocked cell aggregation, decreases cell migration, protected the RBCs from hemolysis caused by LPS, as well as upregulated GILZ expression culminating in reversing the dysfunction of the immune cells, in particular dendritic cells.

Article Snippet: The cyclic peptide inhibitor of integrin α4β1 (azide modified peptide: MePhe-Leu-Asp-Val-Aib-Lys) ( ) control peptides (cyclic MePhe-Ala-Ala-Ala-Aib-Lys), and peptide drug conjugates (PDC), cyclic peptide inhibitor of integrin α4β1 conjugated methylprednisolone (α4β1/Methrol), and control peptide/methrol were synthesized using solid-supported chemistry by InnoPep Inc (San Diego, CA, United States).

Techniques: Expressing, Activation Assay, Migration