Journal: Cancer Research

Article Title: Nerves Stimulate Cross-talk Between Gastric Cancer and Group 3 Innate Lymphoid Cells to Enhance Immunosuppression

doi: 10.1158/0008-5472.CAN-25-3092

Figure Lengend Snippet: Elevated ABHD16A triggered by ACh was associated with poor prognosis in patients with gastric cancer. A, Representative IHC images of ABHD16A expression in human normal gastric tissues and gastric cancer tissues from the TMA with magnifications of 40×, 100×, and 200×. Scale bar for 200×, 100 μm. B–D, Relationship of ABHD16A IHC score to clinical stage ( B ), pT stage ( C ), and lymph node metastasis ( n = 80; D ). E, Cumulative survival curves of patients with gastric cancer with high or low ABHD16A expression based on the TMA. F, IHC images and score of ABHD16A in S100 + or S100 − gastric cancer tissues. Scale bar, 100 μm. G, Procedure for coculture of DRG neurons and gastric cancer cells. H, Western blotting was used to analyze ABHD16A levels in MFC cells cocultured with DRG neurons or DRG neuron–derived CM. GAPDH served as the loading control. I, Expression of HIF1A and ABHD16A in neurotransmitter [HA, dopamine (DA), 5-HT, norepinephrine (NE), ACh]-treated MFC cells and ACh-treated MGC-803 cells. J, ACh concentration was detected in DRG CM by ELISA. K, Tumor volume of orthotopic gastric cancer tumors with or without vagotomy. L, Representative mIF staining images of FOXP3, CD163, CD11b, CD8, PD-L1, and Pan-CK in gastric cancer tissues with high or low expression of ABHD16A. Scale bar, 100 μm. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: GES-1 (RRID: CVCL_EQ22), NCI-N87 (RRID: CVCL_1603), SGC-7901 (RRID: CVCL_0520), and MGC-803 (RRID: CVCL_5334) cells used in this study were purchased from the ATCC.

Techniques: Expressing, Western Blot, Derivative Assay, Control, Concentration Assay, Enzyme-linked Immunosorbent Assay, Staining

Journal: Cancer Research

Article Title: Nerves Stimulate Cross-talk Between Gastric Cancer and Group 3 Innate Lymphoid Cells to Enhance Immunosuppression

doi: 10.1158/0008-5472.CAN-25-3092

Figure Lengend Snippet: IL22 upregulates PD-L1 expression in gastric cancer cells through the UPR IRE1α–XBP1 axis. A, mIF images show the alterations of PD-L1 + tumor cells (purple), CD4 + (green), and CD8 + (red) T cells in orthotopic gastric cancer tumors from control and Abhd16a -knockdown mice following IL22 treatment. Scale bar, 50 μm. B, KEGG pathway enrichment analysis of RNA-seq data of gastric cancer tissues with or without IL22 treatment. C, RT-PCR was used to assess the mRNA expression of key downstream molecules of the UPR branches ( XBP1 , ATF4 , ATF6 ) in control and IL22RA1 -knockdown gastric cancer cells. D, Western blotting analysis of PD-L1 and XBP1s levels in control and IL22RA1 -knockdown MGC-803 cells treated with IL22 (100 μg/L). E, Western blotting detection of PD-L1 and XBP1s levels in XBP1- knockdown MGC-803 cells treated with IL22 and MGC-803 cells treated with IL22 or XBP1s inhibitor (STF083010, 30 μmol/L) in combination with IL22. F, The binding sequence of XBP1 on the CD274 promoter. G and H, ChIP ( G ) and luciferase reporter assay ( H ) showing the transcriptional regulation of CD274 by XBP1s under IL22 stimulation. I, Orthotopic gastric cancer mouse models ( n = 5 per group) were injected with anti-IL22 (200 μg per mouse), anti-CD90.2 antibody (150 μg per mouse), anti-CD90.2 antibody in combination with IL22 (500 ng per mouse), or anti-CD90.2 antibody in combination with XBP1s inhibitors (STF083010, 30 mg/kg) and IL22 for 2 weeks. IHC analysis was used to show IL22, XBP1s, and PD-L1 levels in gastric cancer tissues. Scale bar, 200 μm. J, Tumor volume of orthotopic gastric cancer models under treatments the same as in I . *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, nonsignificant.

Article Snippet: GES-1 (RRID: CVCL_EQ22), NCI-N87 (RRID: CVCL_1603), SGC-7901 (RRID: CVCL_0520), and MGC-803 (RRID: CVCL_5334) cells used in this study were purchased from the ATCC.

Techniques: Expressing, Control, Knockdown, RNA Sequencing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Binding Assay, Sequencing, Luciferase, Reporter Assay, Injection

Journal: JIMD Reports

Article Title: Small Molecules as Alternate Substrates for 3‐Methylglutaconylation

doi: 10.1002/jmd2.70047

Figure Lengend Snippet: Non‐enzymatic reactions of trans‐3MGC‐CoA. The series of non‐enzymatic chemical reactions induced by the accumulation of trans ‐3MGC‐CoA in vivo is depicted. Trans ‐3MGC‐CoA formed as an intermediate in the leucine catabolism pathway or alternatively by the acetyl CoA diversion pathway spontaneously isomerizes to cis ‐3MGC‐CoA. A subsequent intramolecular cyclization reaction generates cis ‐3MGC anhydride and free CoA. 3MGC anhydride has three potential fates, including hydrolysis to yield cis ‐3MGC acid (center), acylation of protein lysine residues (left), or acylation of amino group‐containing small molecules (right).

Article Snippet: 3‐MGC anhydride (4‐methyl‐2H‐pyran‐2,6(3H)‐dione) was from Biosynth Ltd. (UK).

Techniques: In Vivo

Journal: JIMD Reports

Article Title: Small Molecules as Alternate Substrates for 3‐Methylglutaconylation

doi: 10.1002/jmd2.70047

Figure Lengend Snippet: Effect of glycine on the signal intensity of 3MGCylated BSA. (A) Schematic diagram depicting the experimental design employed. “Incubation 1” refers to the incubation of 3MGC‐CoA filtrate with glycine while “Incubation 2” refers to the further incubation of this sample following the addition of BSA. Dashed box represents expected products formed. Control experiments with no glycine added were conducted in parallel. (B) Incubations were conducted as described in panel A and, following incubation, an aliquot of each sample was subjected to SDS‐PAGE and immunoblotting with α‐3MGC IgG. Dotted line indicates that the immunoblot was cut to remove extraneous lanes. (C) Incubations were conducted and processed as in panel B, except 3MGC anhydride was used in lieu of trans ‐3MGC‐CoA filtrate.

Article Snippet: 3‐MGC anhydride (4‐methyl‐2H‐pyran‐2,6(3H)‐dione) was from Biosynth Ltd. (UK).

Techniques: Incubation, Control, SDS Page, Western Blot

Journal: JIMD Reports

Article Title: Small Molecules as Alternate Substrates for 3‐Methylglutaconylation

doi: 10.1002/jmd2.70047

Figure Lengend Snippet: Effect of small molecules on trans ‐3MGC‐CoA‐dependent acylation of BSA. (A) A trans ‐3MGC‐CoA‐containing 3‐MCC reaction filtrate was incubated with indicated amounts of N‐acetylglucosamine or glucosamine for 2 h at 37°C. BSA (0.5 mg/mL) was then added, and the samples incubated for a further 24 h. An aliquot of each sample was then subjected to α‐3MGC IgG immunoblot analysis. (B) A trans ‐3MGC‐CoA‐containing 3‐MCC reaction filtrate was incubated with specified amounts of choline or ethanolamine for 2 h at 37°C. BSA (0.5 mg/mL) was then added, and the samples incubated for a further 24 h. An aliquot of each sample was then subjected to SDS‐PAGE and immunoblotting with α‐3MGC IgG. (C) A trans ‐3MGC‐CoA‐containing 3‐MCC reaction filtrate was incubated with specified amounts of glutathione for 2 h at 37°C. BSA (0.5 mg/mL) was then added, and the samples incubated for a further 24 h. An aliquot of each sample was then subjected to α‐3MGC IgG immunoblot analysis.

Article Snippet: 3‐MGC anhydride (4‐methyl‐2H‐pyran‐2,6(3H)‐dione) was from Biosynth Ltd. (UK).

Techniques: Incubation, Western Blot, SDS Page