Journal: Nature Communications

Article Title: Histone lactylation increases CXCL1 expression for neutrophil infiltration and immune escape in pancreatic cancer

doi: 10.1038/s41467-026-69311-5

Figure Lengend Snippet: A The expression of neutrophils and CD8 + T cells in low- and high-glycolysis clusters, divided by ssGSEA score of glycolysis-related genes, in TCGA and CPTAC PAAD database. Data are presented as box plots showing the median (center line), the first and third quartiles (box bounds), and the whiskers extend to 1.5 times the interquartile range from the box. B The correlation of glycolysis level and neutrophils or CD8 + T cells expression in TCGA ( n = 179 patients) and CPTAC ( n = 140 patients) PAAD database. C Kaplan–Meier plots representing survival probabilities in TCGA-PAAD patients according to the relative level of glycolysis-related or neutrophil-related gene expression. D The diagram illustrating the targets of 2DG and shRNA in the glycolysis process. E CyTOF analysis of tumor-infiltrating immunocytes in subcutaneous tumor models with or without 2DG treatment: tSNE plots showing 12 meta-clusters based on the expression of 41 markers for the immunocytes. F Bar chart of the frequencies of the immune cell subsets in two experimental groups. G A schematic diagram showing the orthotopic PDAC model ( n = 6 mice per group in one experiment) with or without 2DG treatment. H , I Image and weights of the orthotopic tumors in the experimental groups from ( G ) at the end of the experiments. ( n = 6 mice per group). ( J – M ) Tumor-infiltrating neutrophils, CD8 + T cells, GZMB + CD8 + T cells, and PD-1 + CD8 + T cells isolated from the orthotopic tumors were analyzed using flow cytometry: representative graphs of flow cytometry (left panel) and statistical analysis of the cell ratio (right panel) ( n = 6 mice per group). N A schematic diagram showing the orthotopic tumor model in C57BL/6 J mice ( n = 6 mice per group in one experiment) treated with or without anti-Ly6G/anti-CD8 antibody. O , P Image and weights of the orthotopic tumors in the experimental groups from ( N ) at the end of the experiments ( n = 6 mice per group). Q – S Tumor-infiltrating neutrophils, CD8 + T cells, and GZMB + CD8 + T cells isolated from the orthotopic tumors were analyzed using flow cytometry: Representative graphs of flow cytometry (left panel) and statistical analysis of the cell ratio (right panel) ( n = 6 mice per group). T , U Representative luminescence images of the mouse model in ( N ) and statistical analysis of the total flux ( n = 6 mice per group). Data represent mean ± SEM. Statistical analysis was conducted using the two-tailed unpaired Student’s t test ( A, I–M, P–S, and U ), two-tailed Pearson’s correlation analysis ( B ), and log-rank test ( C ). Source data are provided as a Source Data file.

Article Snippet: The treatment schedule is depicted in the figures in the manuscript and described as following: (1) The treatments of vehicle/2DG (800 mg/kg, i.p., once daily) was initiated on day 7 post-implantation and continued for 2 or 8 days; (2) Administration of vehicle/bromosporine (30 mg/kg, i.p., once a day) was initiated on day 9 post-implantation and continued for a total of 11 doses; (3) For combinational treatment, the administration of bromosporine (30 mg/kg, i.p., 3 times a week) or anti-PD-1 antibody (200 μg/mouse, i.p., twice a week, BioXCell, #BE0146)/IgG2a antibody (200 μg/mouse, i.p., twice a week, BioXCell, #BE0089) began on day 13 post-implantation.

Techniques: Expressing, Gene Expression, shRNA, Isolation, Flow Cytometry, Two Tailed Test

Journal: Nature Communications

Article Title: Histone lactylation increases CXCL1 expression for neutrophil infiltration and immune escape in pancreatic cancer

doi: 10.1038/s41467-026-69311-5

Figure Lengend Snippet: A Molecular docking simulated the binding affinities of PCAF for acetyl-CoA and lactyl-CoA. B Co-IP was performed to confirm the interaction between H3K18la and PCAF in PANC1 and KPC cells. PCAF and H3K18la were detected on the same gel. C, D Western blot analysis demonstrates the level of H3K18la in PDAC cells with or without the treatment of PCAF inhibitors (bromosporine or Embelin). Histone H3 and H3K18la blots are from parallel-processed separate gels (size conflict) with samples from the same experiment. E Western blot analysis demonstrates the level of H3K18la in PDAC cells following the knockdown of Pcaf . Histone H3, PCAF, β-actin and H3K18la blots are from parallel-processed separate gels with samples from the same experiment. F Western blots of in vitro histone acetylation or lactylation assay, the samples as indicated. Blots are from parallel-processed separate gels with samples from the same experiment. G , H Relative Cxcl1 RNA and protein levels with the vehicle or bromosporine treatment were analyzed by qRT-PCR and ELISA ( n = 3 independent experiments), respectively. I Relative Cxcl1 RNA levels after Pcaf knockdown were analyzed by qRT-PCR ( n = 3 independent experiments). J A schematic diagram showing the orthotopic tumor construction in C57BL/6 J mice ( n = 5 mice per group in one experiment, 2 × 10 6 KPC-luc cells per mouse) with or without bromosporine treatment. K , L Image and weights of the orthotopic tumors in the experimental groups from ( J ) at the end of the experiments ( n = 5 mice per group). M Lactylation levels of H3K18 and PanK in orthotopic tumors with or without bromosporine treatment were detected by western blot ( n = 4 biologically independent samples per group). Histone H3, H3K18la, and PanKla blots are from parallel-processed separate gels (size conflict) with samples from the same experiment. N Serum CXCL1 levels in mice with or without bromosporine treatment were measured by ELISA ( n = 5 mice per group). O – R Tumor-infiltrating neutrophils, CD8 + T cells, GZMB + CD8 + T cells, and PD-1 + CD8 + T cells isolated from the orthotopic tumors were analyzed using flow cytometry. Representative graphs of flow cytometry (left panel) and statistical analysis of the cell ratio (right panel) ( n = 5 mice per group). S The migratory abilities of neutrophils co-incubated with the culture medium supernatant from shNTC/ Pcaf PDAC cells treated with recombinant CXCL1 were analyzed by counting the penetrated cell numbers ( n = 3 biologically independent samples). T Representative IHC images stained by CXCL1 or H3K18la antibody (scale bar = 100 μm, left panel), correlation analysis of H-scores of CXCL1 and H3K18la in PDAC samples ( n = 21 patients, right panel). Data represent mean ± SEM. Statistical analysis was conducted using the two-tailed unpaired Student’s t test ( G – I , L , and N – S ) and two-tailed Pearson’s correlation analysis ( T ). Unless otherwise indicated, all western blots had three independent experimental repetitions with consistent results. Source data are provided as a Source Data file.

Article Snippet: The treatment schedule is depicted in the figures in the manuscript and described as following: (1) The treatments of vehicle/2DG (800 mg/kg, i.p., once daily) was initiated on day 7 post-implantation and continued for 2 or 8 days; (2) Administration of vehicle/bromosporine (30 mg/kg, i.p., once a day) was initiated on day 9 post-implantation and continued for a total of 11 doses; (3) For combinational treatment, the administration of bromosporine (30 mg/kg, i.p., 3 times a week) or anti-PD-1 antibody (200 μg/mouse, i.p., twice a week, BioXCell, #BE0146)/IgG2a antibody (200 μg/mouse, i.p., twice a week, BioXCell, #BE0089) began on day 13 post-implantation.

Techniques: Binding Assay, Co-Immunoprecipitation Assay, Western Blot, Knockdown, In Vitro, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Isolation, Flow Cytometry, Incubation, Recombinant, Staining, Two Tailed Test

Journal: Nature Communications

Article Title: Histone lactylation increases CXCL1 expression for neutrophil infiltration and immune escape in pancreatic cancer

doi: 10.1038/s41467-026-69311-5

Figure Lengend Snippet: A A schematic diagram showing the subcutaneous tumor model ( n = 6 mice per group in one experiment, 6 × 10 6 PANC02 cells per mouse) treated with bromosporine and anti-PD-1 antibody. B. The tumor volume growth curves of subcutaneous tumors from ( A ). C , D Image and weights of the subcutaneous tumors at the end point of experiments ( n = 6 mice per group). E The statistical analysis of the cell ratio of tumor-infiltrating neutrophils and CD8 + T cells isolated from subcutaneous tumors ( n = 3 mice per group). F Western blot analysis demonstrates H3K18la levels in the subcutaneous tumors ( n = 3 biologically independent samples per group) from ( A ). Histone H3 and H3K18la blots are from parallel-processed separate gels (size conflict) with samples from the same experiment. G Relative serum CXCL1 levels in mice treated with or without bromosporine/anti-PD-1 antibody were measured by ELISA ( n = 6 mice per group). H A schematic diagram showing the combinational treatment schedule for the orthotopic KPC-luc tumor model ( n = 21 mice per group in one experiment, 2 × 10 6 KPC-luc cells per mouse). I , J Image and weights of the orthotopic tumors at the end point of experiments ( n = 6 mice per group). K – N Tumor-infiltrating neutrophils, CD8 + T cells, GZMB + CD8 + T cells, and PD-1 + CD8 + T cells were analyzed using flow cytometry. Representative graphs of flow cytometry (left panel) and statistical analysis of the cell ratio (right panel) ( n = 6 mice per group). O Western blot analysis showing the H3K18la levels in the orthotopic tumors ( n = 3 biologically independent samples per group) from ( H ). Histone H3 and H3K18la blots are from parallel-processed separate gels (size conflict) with samples from the same experiment. P Representative luminescence images and Quantification of radiance intensity of the mouse model in ( H ). Q Survival probability of mice with orthotopically transplanted PDAC ( n = 15 mice per group). R A working model displaying the signaling pathway through which the aerobic glycolysis-mediated Lactate-PCAF-H3K18la-CXCL1 axis modulates the tumor microenvironment in pancreatic cancer, and the scientific basis for the development of a novel therapeutic strategy for PDAC. Data represent mean ± SEM. Statistical analysis was conducted using the two-tailed unpaired Student’s t test ( B , D , E , G , J – N , P ) and the log-rank test ( Q ). Source data are provided as a Source Data file.

Article Snippet: The treatment schedule is depicted in the figures in the manuscript and described as following: (1) The treatments of vehicle/2DG (800 mg/kg, i.p., once daily) was initiated on day 7 post-implantation and continued for 2 or 8 days; (2) Administration of vehicle/bromosporine (30 mg/kg, i.p., once a day) was initiated on day 9 post-implantation and continued for a total of 11 doses; (3) For combinational treatment, the administration of bromosporine (30 mg/kg, i.p., 3 times a week) or anti-PD-1 antibody (200 μg/mouse, i.p., twice a week, BioXCell, #BE0146)/IgG2a antibody (200 μg/mouse, i.p., twice a week, BioXCell, #BE0089) began on day 13 post-implantation.

Techniques: Isolation, Western Blot, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: CRISPR screens in the context of immune selection identify CHD1 and MAP3K7 as mediators of cancer immunotherapy resistance

doi: 10.1016/j.xcrm.2025.102565

Figure Lengend Snippet: Reduced CHD1 and MAP3K7 expression enhances anti-tumor immunity and correlates with clinical response to immune checkpoint blockade (A) Chd1 and Map3k7 dKO B16-F10 tumor cells have a higher frequency of spontaneous rejection. Engraftment rates of NT gRNA-harboring or dKO B16-F10 cells injected subcutaneously into syngeneic C57BL/6 mice at day 9 post-injection. Data represent four independent experiments. Two-sided Fisher’s exact test; ∗ P = 0.015. (B) Chd1 and Map3k7 dKO B16-F10 tumors grow more slowly in vivo . Endpoint tumor mass of successfully engrafted B16-F10 NT gRNA and dKO tumors in mice treated with intraperitoneal anti-PD-1 and anti-CTLA-4 immune checkpoint blockade (ICB) or isotype (Iso.) control. Data represent the mean ± SD from three independent experiments. Unpaired, two-tailed Student’s t test; n = 25 NT NT iso.; n = 18 NT NT ICB; n = 33 dKO iso.; n = 21 dKO ICB. ∗∗∗ P = 0.0002, ∗∗ P = 0.0025 NT NT iso. versus NT NT ICB, ∗∗ P = 0.0094 dKO iso versus dKO ICB, ∗ P = 0.01. (C) Tumor regressions are more frequent in Chd1 and Map3k7 dKO B16-F10 tumors treated with ICB. Subcutaneous tumors were measured using calipers, and tumors with a decrease in volume were considered regressions. Data are pooled from three independent experiments. NT NT ICB versus dKO ICB, two-sided Fisher’s exact test; ∗ P = 0.0176. (D) Improved immunotherapy response in Chd1 and Map3k7 dKO B16-F10 tumors. Growth curves of subcutaneously engrafted B16-F10 NT NT gRNA or dKO tumors ± ICB. Data represent the mean ± SEM and are representative of three independent experiments. Two-way analysis of variance (ANOVA); ∗ P = 0.0094; ∗∗ P = 0.0019. dKO Iso n = 6, dKO ICB n = 5, NT NT Iso n = 4, NT NT ICB n = 6. (E) Heightened anti-tumor immunity in Chd1 and Map3k7 dKO B16-F10 tumors treated with ICB. Immunoprofiling of subcutaneous tumors at the endpoint (days 15–17). Relative abundance of intratumoral conventional CD4 + T cells, CD8 + T cells, regulatory T cells (T regs ), and the CD8 + T cells:T reg ratio, as well as the surface expression of the activation marker CD44 on CD8 + T cells, was assessed by flow cytometry. MFI, mean fluorescence intensity. Data represent the mean ± SD and are pooled from three independent experiments. One-way ANOVA; ∗∗∗∗ P < 0.0001; ∗∗∗ P < 0.0005; ∗∗ P < 0.01; ∗ P < 0.05; ns = not significant. NT NT Iso. n = 22 or n = 23 for CD8 analysis. NT NT ICB n = 20 or n = 19 for Tconv. analysis. dKO Iso. n = 25 or n = 27 for CD8 analysis. dKO ICB n = 17 or n = 16 for CD8:Treg or n = 18 for CD44 analysis. (F) Correlation of CHD1 and MAP3K7 mRNA expression in prostate cancers expressed as log2 fragments per kilobase of transcript per million (FMPK). Spearman’s rank correlation, R = 0.67, P = 1.54e−16. (G) Reduced expression of CHD1 and MAP3K7 in lung cancer and melanoma is associated with clinical response to ICB. Boxplot displaying tumor CHD1 and MAP3K7 mRNA expression (adjusted transcripts per million; adj. TPM) and clinical responses to ICB in patients from the Hartwig Medical Foundation. Significance was assessed using the Wilcoxon signed-rank test, and n denotes the number of patients. Boxplots represent the median and interquartile range (IQR), and whiskers indicate the lowest and highest values within 1.5 × IQR. CB, clinical benefit; NCB, no clinical benefit.

Article Snippet: Anti-human PD-1 Isotype control , BioXCell , Cat.# BP0089; RRID: AB_2894744.

Techniques: Expressing, Injection, In Vivo, Control, Two Tailed Test, Activation Assay, Marker, Flow Cytometry, Fluorescence