Journal: Cell Reports Medicine

Article Title: Disruption of MerTK increases the efficacy of checkpoint inhibitor by enhancing ferroptosis and immune response in hepatocellular carcinoma

doi: 10.1016/j.xcrm.2024.101415

Figure Lengend Snippet: MerTK mediates HCC resistant to anti-PD-L1 treatment (A) Schematic illustrating the establishment of anti-PD-L1-resistant strains in vivo . (B) Tumor growth curves of subcutaneous implantation models in Hepa1-6 and Res1-6 strains treated with anti-PD-L1 (aPD-L1) or IgG. (C) Survival of orthotopic implantation models of Hepa1-6 and Res1-6 strains treated with aPD-L1 or IgG. (D) Proteomic sequencing analysis was used to analyze the differently expressed proteins in Hepa1-6 and Res1-6. The top 20 differently expressed proteins are presented in a heatmap, including 10 upregulated and 10 downregulated proteins. (E) Western blot of p-MerTK, MerTK, and β-actin in Hepa1-6, Res1-6, HCA-1, and Res-CA1 strains. (F) IHC staining of p-MerTK and MerTK in Hepa1-6 and Res1-6 subcutaneous tumor tissues. (G) Locations of sensitive and resistant tumors were exhibited by MRI, and AFP values before and after anti-PD-L1/PD-1 therapy between patient in sensitive group and patient in resistant group. IHC staining of MerTK expression in HCC tissues from sensitive and resistant patients received anti-PD-1/PD-L1 therapy. (H and I) Subcutaneous xenograft mouse model of Res1-6, Res1-6-sh-MerTK, Hepa1-6, and Hepa1-6-OE-MerTK strains treated with anti-PD-L1 or IgG. When the tumor volume approximately reached 100 mm 3 in size, tumor volume was measured every 3 days. After 25 days of treatment, mice were sacrificed. Shown are tumor appearance and tumor growth curves. All results are shown as mean ± SEM (n = 5). One- or two-way ANOVA was used to analyze the data; ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.

Article Snippet: For treatment with antibody, 200 μg mouse anti-PD-L1 antibody (BE0361, Bio X Cell) or IgG control (BE0089, Bio X Cell) was injected intraperitoneally (i.p.) twice a week.

Techniques: In Vivo, Sequencing, Western Blot, Immunohistochemistry, Expressing

Journal: Cell Reports Medicine

Article Title: Disruption of MerTK increases the efficacy of checkpoint inhibitor by enhancing ferroptosis and immune response in hepatocellular carcinoma

doi: 10.1016/j.xcrm.2024.101415

Figure Lengend Snippet: MerTK promotes anti-PD-L1 resistance by suppressing ferroptosis in HCC (A) All the differential genes between Hepa1-6 and Res1-6 cells were analyzed using KEGG pathway analysis using the functional gene sets in MSigDB (literature vs. databases containing signaling pathways). (B) Fluorescence detection of lipid ROS by C11-BODIPY (left) and statistical analysis of relative lipid ROS fluorescence signal (right). (C) Fluorescence detection of dead cells by SYTOX Green (left) and statistical analysis of percentage dead cells (right). (D–F) Cell viability of Hepa1-6, Hepa1-6-OE-MerTK, Res1-6, and Res1-6-sh-MerTK strains treated with erastin (5.0 μM) in cocultured condition (left) and statistical analysis of cell survival rate in each time point (right). (G–J) In subcutaneous xenograft mouse model, the statistical analysis of relative lipid ROS and MDA content in Hepa1-6, Hepa1-6-OE-MerTK, Res1-6, and Res1-6-sh-MerTK strains treated with anti-PD-L1 or IgG. All results are shown as mean ± SEM (n = 5). One- or two-way ANOVA was used to analyze the data; ∗∗p < 0.01 and ∗∗∗p < 0.001.

Article Snippet: For treatment with antibody, 200 μg mouse anti-PD-L1 antibody (BE0361, Bio X Cell) or IgG control (BE0089, Bio X Cell) was injected intraperitoneally (i.p.) twice a week.

Techniques: Functional Assay, Protein-Protein interactions, Fluorescence

Journal: Cell Reports Medicine

Article Title: Disruption of MerTK increases the efficacy of checkpoint inhibitor by enhancing ferroptosis and immune response in hepatocellular carcinoma

doi: 10.1016/j.xcrm.2024.101415

Figure Lengend Snippet: Suppression of SLC7A11 sensitizes anti-PD-L1 treatment caused by MerTK (A) Volcano plot of differentially expressed proteins in protein mass spectrometry. (B) Western blot of SLC7A11 expression in Hepa1-6, Res1-6, HCA-1, and Res-CA1. (C) IHC staining of SLC7A11 in Hepa1-6, Res1-6, HCA-1, and Res-CA1 subcutaneous tumor tissues. (D) IHC staining of SLC7A11 expression in HCC tissues from sensitive and resistant patients with anti-PD-1/PD-L1 therapy, and statistical analysis. (E) The correlation between MerTK and SLC7A11 expression in tumor tissues from HCC patients received anti-PD-1/PD-L1 therapy, Pearson product-moment correlation coefficients and p values are shown. (F and G) (F) Schematic illustrating the procedure of anti-PD-L1 or IgG treatment in Res1-6 and Res1-6-shSLC7A11 subcutaneous tumor model, and (G) the representative images of subcutaneous tumor in different groups. (H) Statistical analysis of tumor growth curves. (I and J) Western blot analysis of p-MerTK, MerTK, SLC7A11, and β-actin expression in Hepa1-6, Hepa1-6-OE-MerTK, Res1-6, and Res1-6-shMerTK subcutaneous tumor treated with anti-PD-L1 or IgG. All results are shown as mean ± SEM (n = 5). One- or two-way ANOVA was used to analyze the data; ∗∗p < 0.01 and ∗∗∗p < 0.001.

Article Snippet: For treatment with antibody, 200 μg mouse anti-PD-L1 antibody (BE0361, Bio X Cell) or IgG control (BE0089, Bio X Cell) was injected intraperitoneally (i.p.) twice a week.

Techniques: Mass Spectrometry, Western Blot, Expressing, Immunohistochemistry

Journal: Cell Reports Medicine

Article Title: Disruption of MerTK increases the efficacy of checkpoint inhibitor by enhancing ferroptosis and immune response in hepatocellular carcinoma

doi: 10.1016/j.xcrm.2024.101415

Figure Lengend Snippet: MerTK induces anti-PD-L1 resistance by favoring a protumor microenvironment (A) Relationship between overall survival and CTL levels in HCC patients with low and high MerTK gene copy numbers. (B) The correlations between the mRNA expression levels of MerTK and cytotoxic CD8 + T cells. (C) The representative image of HCC tissue stained with MerTK (red), CD8 (gold), CD11b (purple), CD15 (green), and CD14 (pink). (D–G) (D) The percentage statistical analysis of CD8 + T cells (E) MDSCs, (F) gMDSCs, and (G) mMDSCs in tumor tissues. (H) The correlation analysis between the expression of MerTK and the enrichment of MDSCs. (I and J) T-distributed stochastic neighbor embedding (t-SNE) plot of tumor-infiltrating leukocytes overlaid with color-coded clusters and the frequency of clusters of the indicated immune cell subsets, including CD3 + T cells, CD8 + T cells, IFNγ + CD8 + T cells, CD4 + T cells, CD11b + cells, and MDSCs in Hepa1-6, Hepa1-6-OE-MerTK, Res1-6, and Res1-6-shMerTK subcutaneous tumor model treated with anti-PD-L1 or IgG (left) and the statistical analysis (right). All results are shown as mean ± SEM (n = 5). One- or two-way ANOVA was used to analyze the data; ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.

Article Snippet: For treatment with antibody, 200 μg mouse anti-PD-L1 antibody (BE0361, Bio X Cell) or IgG control (BE0089, Bio X Cell) was injected intraperitoneally (i.p.) twice a week.

Techniques: Expressing, Staining

Journal: Cell Reports Medicine

Article Title: Disruption of MerTK increases the efficacy of checkpoint inhibitor by enhancing ferroptosis and immune response in hepatocellular carcinoma

doi: 10.1016/j.xcrm.2024.101415

Figure Lengend Snippet: Inhibition of MerTK promotes ferroptosis and increases the efficacy of PD-L1 antibody in resistant HCC (A) Cell viability (percentage) analysis of Res1-6 cells following MerTK inhibitors (sitravatinib, UNC5293, UNC2541, and UNC1267) at different concentrations (0, 0.1, 0.5, 1.0, 2.0, 4.0, 8.0, 16.0, and 32.0 μM). (B) Cell death detection by propidium iodide (PI) staining (left) and statistical analysis (right). (C) Fluorescence detection of lipid ROS by C11-BODIPY (left) and statistical analysis of relative lipid ROS fluorescence signal (right). (D) Fluorescence detection of dead cells by SYTOX Green (left) and statistical analysis of percentage dead cells (right). (E) The representative images of subcutaneous tumor in Res1-6 strains were treated with IgG, sitravatinib, anti-PD-L1 or their combination (left) and the statistical analysis of tumor growth curves (right). (F) The representative images of orthotopic tumor in Res1-6 strains were treated with IgG, sitravatinib, anti-PD-L1 or their combination (left) and the statistical analysis of survival curves (right). All results are shown as mean ± SEM (n = 5). One- or two-way ANOVA was used to analyze the data; ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.

Article Snippet: For treatment with antibody, 200 μg mouse anti-PD-L1 antibody (BE0361, Bio X Cell) or IgG control (BE0089, Bio X Cell) was injected intraperitoneally (i.p.) twice a week.

Techniques: Inhibition, Staining, Fluorescence

Journal: Cell Reports Medicine

Article Title: Disruption of MerTK increases the efficacy of checkpoint inhibitor by enhancing ferroptosis and immune response in hepatocellular carcinoma

doi: 10.1016/j.xcrm.2024.101415

Figure Lengend Snippet: Sitravatinib combined with PD-L1 antibody increases ferroptosis and reduces MDSC infiltration in HCC microenvironment (A and B) (A) T-SNE plot of tumor-infiltrating leukocytes overlaid with color-coded clusters and the frequency of clusters of the indicated immune cell subsets, including CD3 + T cells, CD8 + T cells, IFNγ + CD8 + T cells, CD4 + T cells, CD11b + cells, and MDSCs in Res1-6 strains treated with IgG, sitravatinib, anti-PD-L1, or their combination and (B) the statistical analysis. (C) Western blot analysis of p-MerTK, MerTK, SLC7A11, and β-actin expression in different groups. (D) The representative imagines of IHC staining of p-MerTK, MerTK, and Ki-67 from subcutaneous tumors treated with IgG, sitravatinib, anti-PD-L1 or their combination. Scale bar: 100 μm. (E–H) In subcutaneous xenograft mouse model, the statistical analysis of relative lipid ROS and MDA content in Res1-6 and Res-CA1 strains treated with IgG, sitravatinib, anti-PD-L1, or their combination. All results are shown as mean ± SEM (n = 5). One- or two-way ANOVA was used to analyze the data; ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.

Article Snippet: For treatment with antibody, 200 μg mouse anti-PD-L1 antibody (BE0361, Bio X Cell) or IgG control (BE0089, Bio X Cell) was injected intraperitoneally (i.p.) twice a week.

Techniques: Western Blot, Expressing, Immunohistochemistry

Journal: Cell Reports Medicine

Article Title: Disruption of MerTK increases the efficacy of checkpoint inhibitor by enhancing ferroptosis and immune response in hepatocellular carcinoma

doi: 10.1016/j.xcrm.2024.101415

Figure Lengend Snippet:

Article Snippet: For treatment with antibody, 200 μg mouse anti-PD-L1 antibody (BE0361, Bio X Cell) or IgG control (BE0089, Bio X Cell) was injected intraperitoneally (i.p.) twice a week.

Techniques: Virus, Recombinant, Red Blood Cell Lysis, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Isolation, Multiple Displacement Amplification, Expressing, shRNA, Software

Journal: Frontiers in Neuroscience

Article Title: Association Study of Alcohol Dehydrogenase and Aldehyde Dehydrogenase Polymorphism With Alzheimer Disease in the Taiwanese Population

doi: 10.3389/fnins.2021.625885

Figure Lengend Snippet: Background demographic distribution and frequency of the genotype in the pilot study.

Article Snippet: Plasma ADH1C level was determined using human ADH1C ELISA kit (catalog #MBS2889930, MyBioSource, San Diego, CA, United States) and monitored spectrophotometrically at 450 nm on a multifunctional microplate reader (Tecan infinite 200) by following the manufacturer’s instructions.

Techniques:

Journal: Frontiers in Neuroscience

Article Title: Association Study of Alcohol Dehydrogenase and Aldehyde Dehydrogenase Polymorphism With Alzheimer Disease in the Taiwanese Population

doi: 10.3389/fnins.2021.625885

Figure Lengend Snippet: Associations of the candidate SNPs with AD in the pilot study.

Article Snippet: Plasma ADH1C level was determined using human ADH1C ELISA kit (catalog #MBS2889930, MyBioSource, San Diego, CA, United States) and monitored spectrophotometrically at 450 nm on a multifunctional microplate reader (Tecan infinite 200) by following the manufacturer’s instructions.

Techniques: Variant Assay

Journal: Frontiers in Neuroscience

Article Title: Association Study of Alcohol Dehydrogenase and Aldehyde Dehydrogenase Polymorphism With Alzheimer Disease in the Taiwanese Population

doi: 10.3389/fnins.2021.625885

Figure Lengend Snippet: Background demographic distribution and frequency of the genotype in the extension study.

Article Snippet: Plasma ADH1C level was determined using human ADH1C ELISA kit (catalog #MBS2889930, MyBioSource, San Diego, CA, United States) and monitored spectrophotometrically at 450 nm on a multifunctional microplate reader (Tecan infinite 200) by following the manufacturer’s instructions.

Techniques:

Journal: Frontiers in Neuroscience

Article Title: Association Study of Alcohol Dehydrogenase and Aldehyde Dehydrogenase Polymorphism With Alzheimer Disease in the Taiwanese Population

doi: 10.3389/fnins.2021.625885

Figure Lengend Snippet: Associations of the candidate SNPs with AD in the extension study.

Article Snippet: Plasma ADH1C level was determined using human ADH1C ELISA kit (catalog #MBS2889930, MyBioSource, San Diego, CA, United States) and monitored spectrophotometrically at 450 nm on a multifunctional microplate reader (Tecan infinite 200) by following the manufacturer’s instructions.

Techniques: Variant Assay

Journal: Frontiers in Neuroscience

Article Title: Association Study of Alcohol Dehydrogenase and Aldehyde Dehydrogenase Polymorphism With Alzheimer Disease in the Taiwanese Population

doi: 10.3389/fnins.2021.625885

Figure Lengend Snippet: ADH1C level in AD and control. Plasma ADH1C level was higher in AD than in control (A) . (B) there was a marginal interaction effect between AD and rs2241894 genotype on plasma ADH1C level ( p = 0.04), in which those carried minor allele T had lower ADH1C level in AD patients but higher ADH1C level in the controls.

Article Snippet: Plasma ADH1C level was determined using human ADH1C ELISA kit (catalog #MBS2889930, MyBioSource, San Diego, CA, United States) and monitored spectrophotometrically at 450 nm on a multifunctional microplate reader (Tecan infinite 200) by following the manufacturer’s instructions.

Techniques: Control, Clinical Proteomics