integrin inhibitors rgd peptides  (Janssen)

Journal: Nature Microbiology

Article Title: Peptostreptococcus anaerobius mediates anti-PD1 therapy resistance and exacerbates colorectal cancer via myeloid-derived suppressor cells in mice

doi: 10.1038/s41564-024-01695-w

Figure Lengend Snippet: a , Representative images of MDSCs that migrated to the lower chamber and quantification of MDSC migration towards CRC-cell-conditioned medium pre-incubated with vehicle or P. anaerobius ( n = 3 biologically independent samples). Scale bars, 100 μm. b , Cytokine array analysis (36 cytokines) of conditioned medium from Caco-2 cells treated with P. anaerobius or vehicle (left). Fold change in cytokine levels after P. anaerobius treatment (right). Red box, significantly changed cytokines; NC, negative control. c , Levels of CXCL1 secreted from Caco-2 and MC38 cells treated with vehicle or P. anaerobius (left). Levels of serum CXCL1 and mRNA expression of CXCL1 in the colon of Apc min/+ mice (right). d , Integrin α 2 β 1 knockdown in Caco-2 cells abrogated P. anaerobius -induced CXCL1 secretion (left) and migration of MDSCs (right). e , RGDS peptide treatment of MC38 cells abrogated P. anaerobius -induced CXCL1 secretion (left) and migration of MDSCs (right). f , Treatment with JSH-23 abolished P. anaerobius -induced CXCL1 secretion (left) and MDSCs migration (right). c – f , n = 3 biologically independent samples. g – j , AOM-induced CRC mice were treated with RGDS. g , Tumour load of the mice; n = 8 (PBS), 11 (PA), 7 (PBS + RGDS) and 10 (PA + RGDS) mice. h , Serum levels of CXCL1; n = 5 (PBS and PA) and 4 (PBS + RGDS and PA + RGDS). i , Percentage of MDSCs in the colonic lamina propria of the mice (left). Immunofluorescence images (middle) and quantification of CD11b + Gr-1 + cells in mouse tumours (right). Scale bars, 25 μm. j , Mean fluorescence intensity (MFI) of IFN-γ + CD8 + and IFN-γ + CD4 + T cells in colonic lamina propria. i , j , n = 7 (PBS and PBS + RGDS) and 9 (PA and PA + RGDS) mice. k – m , MC38 allograft model mice were treated with RGDS peptide and anti-PD1. k , Tumour growth curves (left) and tumour weight (right); n = 7 (PBS + anti-PD1) and 8 (all other groups) mice. l , CXCL1 levels in tumour tissue lysates; n = 5 (PBS + IgG, PBS + anti-PD1 and PBS + anti-PD1 + RGDS), 8 (PA + IgG), 7 (PA + anti-PD1) and 3 (PA + anti-PD1 + RGDS) mice. m , Percentage of MDSCs (left) and IFN-γ + CD8 + T cells (right) in tumours; n = 7 (PBS + anti-PD1) and 8 (all other groups) mice. a , c – m , Data are the mean ± s.e.m. P values were calculated using a one-way ANOVA, followed by Tukey’s post-hoc test ( a , d – j ); an unpaired two-tailed Student’s t -test ( c ); a one-way ANOVA, followed by Fisher’s least significant difference test ( k – m ) or a two-way ANOVA, followed by Bonferroni’s post-hoc test ( k ).

Article Snippet: For the inhibitor-treatment experiments, cells were pretreated with 30 μM NF-κB inhibitor JSH-23 (MCE) or 100 μM integrin α 2 β 1 inhibitor RGDS peptide (MCE) for 24 h and then treated with P. anaerobius .

Techniques: Migration, Incubation, Negative Control, Expressing, Knockdown, Immunofluorescence, Fluorescence, Two Tailed Test

Journal: Nature Microbiology

Article Title: Peptostreptococcus anaerobius mediates anti-PD1 therapy resistance and exacerbates colorectal cancer via myeloid-derived suppressor cells in mice

doi: 10.1038/s41564-024-01695-w

Figure Lengend Snippet: a . Validation of integrin α 2/ β 1 knock down efficiency by western blot in HCT116 and Caco-2 cells (up) and Integrin α 2 β 1 knockdown in HCT116 cells abrogated P. anaerobius-induced CXCL1 secretion and migration of MDSCs (down). b . Antibody blocking of integrin α 2/ β 1 suppressed P. anaerobius -induced NF- κ B activation in HCT116 and MC38 cells (up) and RGDS peptide treatment in HCT116 cells abrogated P. anaerobius -induced CXCL1 secretion and migration of MDSCs (down). a-b n = 3 biologically independent samples. c . P. anaerobius promoted the nuclear translocation of NF-κB p65 and phospho-p65 in CRC cells. d . Induction of NF-κB activation by P. anaerobius was abolished by JSH-23 in HCT116, Caco-2 and MC38 cells. Three independent experiments were repeated with similar results in a–d. e . Treatment with JSH-23 abolished P. anaerobius -induced CXCL1 secretion (left) and MDSCs migration (right) in conditioned medium from HCT116 cell (n = 3 biologically independent samples). f . Schematic diagram of RGDS treatment in mice. RGDS abrogated P. anaerobius -induced colorectal tumorigenesis. g . Representative flow plots of IFNγ + CD8 + T cells and IFNγ + CD4 + T cells in colonic LP cells. h . Schematic diagram of MC38 allograft model. RGDS peptide reversed P. anaerobius -mediated anti-PD1 resistance in MC38 allografts. Representative images of tumours after sacrificing. i . Percentage of G-MDSCs in tumours were determined by flow cytometry analysis. 6-8 mice were used in each group including IgG (n = 8), a-PD1 (n = 6), a-PD1 + RGDS (n = 7), PA+ IgG (n = 8), PA+ a-PD1 (n = 8), PA+ a-PD1 + RGDS (n = 7). j . Representative tumour flow plots of IFNγ + CD8 + T cells in tumours. k . Integrin α 2/ β 1 blockade abolished the effect of P. anaerobius on NF- κ B pathway in MC38 allograft tumours (n = 4 biologically independent samples). Data are presented as mean ± SEM. P values were calculated by one-way ANOVA followed by Tukey’s post-hoc test ( e, i ).

Article Snippet: For the inhibitor-treatment experiments, cells were pretreated with 30 μM NF-κB inhibitor JSH-23 (MCE) or 100 μM integrin α 2 β 1 inhibitor RGDS peptide (MCE) for 24 h and then treated with P. anaerobius .

Techniques: Biomarker Discovery, Knockdown, Western Blot, Migration, Blocking Assay, Activation Assay, Translocation Assay, Flow Cytometry

Journal: mBio

Article Title: Mindin Activates Autophagy for Lipid Utilization and Facilitates White Spot Syndrome Virus Infection in Shrimp

doi: 10.1128/mbio.02919-22

Figure Lengend Snippet: Integrin is essential for Mindin-enhanced WSSV infection. (A) Influence of integrin signaling inhibition by using RGD peptide on Mindin-induced autophagy. RGD peptide was administered at the dose of 5 μg/shrimp to block integrin signaling. rMindin injection (5 μg) was performed at 2 h after RGD peptide application. GABARAP expression in hemocytes was detected another 6 h later. (B and C) Influence of integrin signaling inhibition on Mindin-induced variation of LDs and the ATP level. rMindin injection was performed at 2 h after RGD peptide application. The hepatopancreas was collected to observe the LDs another 12 h later (B) or to detect the ATP level another 6 h later (C). Bar, 10 μm. Quantification of LD diameter was performed by ImageJ software. (D) Influence of integrin signaling inhibition on Mindin-enhanced WSSV entry. FITC-labeled virions (10 6 copies) were injected into shrimp hemocoels together with rMindin or the control tag (5 μg) at 2 h after RGD peptide application. WSSV entry was detected using flow cytometry 1 h later. (E) Influence of integrin signaling inhibition on Mindin-enhanced WSSV replication. Injection of rMindin and WSSV inoculum (5 × 10 5 copies) was performed at 2 h after RGD peptide application. WSSV load (left panel) and VP28 expression (right panel) in gills were detected 24 h later. (F) Effect of integrin knockdown on Mindin-enhanced WSSV infection. Injection of rMindin and WSSV inoculum (5 × 10 5 copies) was performed at 48 h after dsRNA injection. WSSV load (left panel) and VP28 expression (right panel) in gills were detected 24 h later. All bar chart data are shown as the mean ± SD from three or four replicates. ***, P < 0.001, and ns, no significant difference, Student’s t test. All Western blotting images, microscopy images, and flow cytometry images are representative of three independent replicates. The numbers in blotting panels indicate the relative band intensities (VP28/β-actin).

Article Snippet: The autophagy inhibitor (3-methyladenine [3-MA]; S2767), the fatty acid oxidation inhibitor (trimetazidine [TMZ]; S4543), the ATP synthase inhibitor (oligomycin A; S1478), and the integrin inhibitor (RGD peptide; S8008) were purchased from Selleck Chemicals (Houston, TX, USA).

Techniques: Infection, Inhibition, Blocking Assay, Injection, Expressing, Software, Labeling, Control, Flow Cytometry, Knockdown, Western Blot, Microscopy

Journal: mBio

Article Title: Mindin Activates Autophagy for Lipid Utilization and Facilitates White Spot Syndrome Virus Infection in Shrimp

doi: 10.1128/mbio.02919-22

Figure Lengend Snippet: Model of the mechanism revealed in this study. Mindin interacts with integrin to induce autophagy. Autophagy regulates lipid droplet (LD) degradation to generate free fatty acids (FFAs) and produce ATP, which is essential for the entry and replication of WSSV. Moreover, the monopolization of lysosome for LD degradation may allow the virions to escape from lysosomal elimination.

Article Snippet: The autophagy inhibitor (3-methyladenine [3-MA]; S2767), the fatty acid oxidation inhibitor (trimetazidine [TMZ]; S4543), the ATP synthase inhibitor (oligomycin A; S1478), and the integrin inhibitor (RGD peptide; S8008) were purchased from Selleck Chemicals (Houston, TX, USA).

Techniques:

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: STAT3 is Activated by CTGF-mediated Tumor-stroma Cross Talk to Promote HCC Progression

doi: 10.1016/j.jcmgh.2022.09.006

Figure Lengend Snippet: Recombinant CTGF treatment upregulated IL-6 family cytokine expression in stromal cell lines via integrin/NF-kB signaling. LX-2, THP-1, MTA, and TMNK-1 cells were serum-starved for 16 hours and then treated with 5 nM recombinant CTGF protein. THP-1 cells had been pretreated with 100 ng/mL phorbol 12-myristate 13-acetate (PMA) to induce macrophage differentiation for 48 hours before starvation. Total protein was collected at the indicated time points after the addition of recombinant CTGF protein. Total RNA was extracted 6 hours after incubation with recombinant CTGF protein. Protein and gene expression after recombinant CTGF treatment ( A-C ). Cells were treated with recombinant CTGF alone ( A ) or pretreated with 1 mM RGDS peptide ( B ) or 50 ng/mL SN-50 ( C ) for 2 hours before the addition of recombinant CTGF protein. n = 4 ( A, B ) or n = 3 ( C ). ∗ P < .05.

Article Snippet: The pan-integrin inhibitor RGDS peptide was purchased from R&D Systems. siRNA against STAT3, gp130, and CTGF and control siRNA were purchased from Thermo Fisher Scientific and transfected into the cells using Lipofectamine RNAiMAX (Thermo Fisher Scientific) according to the Lipofectamine reverse transfection protocol.

Techniques: Recombinant, Expressing, Incubation, Gene Expression