Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Mutant p53 epigenetically rewires CXCL10 to promote CD8⁺ T-cell infiltration and enhance the anti-PD-1 response in advanced prostate cancer

doi: 10.1186/s13046-026-03672-z

Figure Lengend Snippet: Mutp53 promotes immunotherapy responsiveness through CXCL10-CXCR3-mediated recruitment and activation of CD8 + T cells. a-b Heatmap ( a ) and qRT-PCR analysis ( b ) of immune-related chemokine expression in subcutaneous WTp53 and Mutp53 tumors, showing significant upregulation of Cxcl10 together with Cxcl1 , Cxcl2 , Cxcl12 , Ccl2 , and Cxcl9 in Mutp53 tumors compared with WTp53 tumors. c Correlation analysis of CXCL10 expression with immune-related signaling pathways in the IMvigor210 urothelial carcinoma cohort treated with immune checkpoint blockade, revealing strong positive associations with IFN-γ-related signatures and the antigen presentation machinery (APM). d Representative immunohistochemical (IHC) staining of CXCL10 in WTp53 and Mutp53 tumors. e-f Immunoblot analysis ( e ) and quantitative densitometry ( f ) of CXCL10 protein expression in WTp53 and Mutp53 tumor cell lines and corresponding tumor tissues. g Transwell coculture assays demonstrating enhanced chemotaxis of CD8 + T cells toward Mutp53-derived tumor cells compared with WTp53-derived tumor cells ( n = 3). h Clinical association between intratumoral CXCL10 expression and immune checkpoint blockade (ICB) benefit in the IMvigor210 cohort, with higher CXCL10 levels predicting improved therapeutic response. i Schematic illustration of the in vivo experimental design for pharmacological disruption of the CXCL10-CXCR3 axis using the CXCR3 antagonist AMG-487 during anti-PD-1 treatment, with RM-1 tumors established by subcutaneous injection of 1 × 10⁶ cells. j-l Representative tumor images ( j ), tumor growth curves ( k ), and endpoint tumor weights ( l ) of WTp53- and Mutp53-bearing mice treated with anti-PD-1 therapy in the presence or absence of AMG-487 ( n = 5 per group). CXCR3 blockade markedly attenuated the therapeutic efficacy of PD-1 blockade, with a substantially stronger reversal observed in Mutp53 tumors. m Serum biochemical analyses evaluating liver and kidney function, including alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea (BUN), and creatinine (CREA), showing no significant differences among treatment groups, indicating minimal systemic toxicity of AMG-487. n Multiplex immunofluorescence staining of tumor sections for CD8 (green), granzyme B (GZMB; red), and nuclei (DAPI; blue), revealing that anti-PD-1 therapy markedly increased infiltration of GZMB+CD8 + T cells in Mutp53 tumors, whereas CXCR3 inhibition significantly impaired this recruitment. o Quantification of GZMB+CD8 + T cells across treatment groups, confirming that the enrichment induced by PD-1 blockade in Mutp53 tumors was largely abolished by AMG-487 treatment. Note: Data are presented as the mean ± SD. Statistical significance was assessed using one-way or two-way ANOVA with Tukey’s multiple-comparison test, as appropriate. p < 0.05, p < 0.01, * p < 0.001; ns, not significant

Article Snippet: Animals received intraperitoneal administration of anti-PD-1 antibody (10 mg/kg; BioXcell, Lebanon, NH, USA) and/or the CXCR3 antagonist AMG487 (5 mg/kg; MedChemExpress, Shanghai, China) according to predefined schedules.

Techniques: Activation Assay, Quantitative RT-PCR, Expressing, Protein-Protein interactions, Immunopeptidomics, Immunohistochemical staining, Immunohistochemistry, Western Blot, Chemotaxis Assay, Derivative Assay, Clinical Proteomics, In Vivo, Disruption, Injection, Drug discovery, Multiplex Assay, Immunofluorescence, Staining, Inhibition, Comparison

Journal: Bioactive Materials

Article Title: Microenvironment-educated MSC-EVs loaded injectable smart hydrogel for targeting senescent nucleus pulposus cells and inhibiting ferroptosis against intervertebral disc degeneration

doi: 10.1016/j.bioactmat.2026.02.030

Figure Lengend Snippet: Targeting Capability of D-EVs to Senescent NPCs is Mediated by the CXCL10-CXCR3 Axis. (A) Schematic diagram of RNA sequencing for senescent NPCs treated with D-EVs or not. (B-C) Volcano plots and heatmaps displaying differential gene expression in senescent NPCs treated with D-EVs or not. (D) A Venn diagram illustrates overlap between DEGs identified in senescent NPCs treated with D-EVs and key gene databases for cytokine-cytokine receptor interactions. (E) A Venn diagram illustrates overlap between DEGs identified in D-MSCs and chemokine signaling pathway key gene databases. (F-G) Network analysis of the hub genes among the above total DEGs. A protein-protein interaction network was created in the STRING database, while the Cytoscape software was used to determine the hub genes in the network. (H) GO analysis confirming enrichment of terms related to senescent NPCs treated with D-EVs or not in the BP categories. (I) GSEA analysis of cytokine-cytokine receptor interactions in D-EVs group versus TBHP group. (J) Western blot analysis confirmed the expression of CXCR3 in Control and senescent NPCs, and (S) quantitative analysis. (K) Western blot analysis confirmed the expression of CXCL10 in N-EVs and D-EVs, and (R) quantitative analysis. (L) Protein binding analysis of CXCR3 and CXCL10 by PyMOL. (M) Confocal analysis of the uptake of pHrodo-labeled EVs in senescent NPCs following the treatments of N-EVs, D-EVs with CXCL10 knockout/anti-CXCR3, or not, and (Q) quantitative analysis. (N) In vivo tracking of PKH26-labeled CXCL10 knockout D-EVs. (O) Flow cytometry showing uptake of different EVs with CXCL10 knockout/anti-CXCR3 or not by senescent NPCs, and (P) quantitative analysis. The data were presented as mean ± SD. n = 3, ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: After blocked with 5% non-fat milk for 2 h at room temperature, the membranes were incubated with primary antibodies against GAPDH (1:5000, 104941-AP, Proteintech), TSG101 (1:1000, DF8427, Affinity), CD9 (1:1000, AF5139, Affinity), CD63 (1:2000, 25682-1-AP, Proteintech), Calnexin (1:5000, 10427-2-AP, Proteintech), GM130 (1:20000, 11308-1-AP, Proteintech), CXCR3 (1:5000, 26756-1-AP, Proteintech), CXCL10 (1:2000, 10937-1-AP, Proteintech), MMP3 (1:2000, 17873-1-AP, Proteintech), ADAMTS5 (DF13268, Affinity), P16 (AF5484, Affinity), P21 (10355-1-AP, Proteintech), GPX4 (1:1000, 381958, Zen-bio), SLC7A11 (1:1000, 26864-1-AP, Proteintech), ACSL4 (1:5000, 22401-1-AP, Proteintech) and Tubulin (1:10000, T40103 , Abmart) overnight at 4 °C.

Techniques: RNA Sequencing, Gene Expression, Software, Western Blot, Expressing, Control, Protein Binding, Labeling, Knock-Out, In Vivo, Flow Cytometry

Journal: Nutrients

Article Title: Docosahexaenoic Acid Attenuates Visceral Pain by Suppressing Spinal CXCL10/CXCR3/ERK Signaling

doi: 10.3390/nu18071113

Figure Lengend Snippet: TNBS-induced upregulation of CXCR3 in spinal neurons. ( A ) TNBS significantly increased CXCR3 mRNA expression in the spinal cord (L6-S1), as determined by qPCR. **, p < 0.01 vs. NS. Student’s t -test, n = 6. ( B ) Western blot analysis and quantification showing increased CXCR3 protein expression in the spinal cord (L6-S1) of TNBS-treated mice compared with NS controls. **, p < 0.01 vs. NS. Student’s t -test. ( C ) CXCR3 (red) is predominantly colocalized with the neuronal marker NeuN (green). ( D ) CXCR3 (red) shows minimal colocalization with the astrocytic marker GFAP (green). ( E ) CXCR3 (red) shows minimal colocalization with the microglial marker IBA-1 (green).

Article Snippet: Membranes were blocked and incubated overnight at 4 °C with primary antibodies against CXCR3 (1:200, rabbit, Boster (Pleasanton, CA, USA)), pERK (1:1000, rabbit, Cell Signaling (Danvers, MA, USA)), and ERK (1:1000, rabbit, Cell Signaling).

Techniques: Expressing, Western Blot, Marker

Journal: Nutrients

Article Title: Docosahexaenoic Acid Attenuates Visceral Pain by Suppressing Spinal CXCL10/CXCR3/ERK Signaling

doi: 10.3390/nu18071113

Figure Lengend Snippet: Intrathecal CXCR3 antagonist attenuates TNBS-induced visceral pain and spinal ERK activation. ( A ) Intrathecal administration of the CXCR3 antagonist NBI-74330 (20 μg), but not 2 μg, significantly attenuated TNBS-induced visceral hypersensitivity at 0.5 h and 1 h after injection. *, p < 0.05; **, p < 0.01 vs. TNBS + vehicle, n = 6. ( B , C ) Western blot analysis and quantification showing that intrathecal NBI-74330 (20 μg) markedly reduced TNBS-induced ERK phosphorylation in the spinal dorsal horn. *, p < 0.05 vs. TNBS + vehicle.

Article Snippet: Membranes were blocked and incubated overnight at 4 °C with primary antibodies against CXCR3 (1:200, rabbit, Boster (Pleasanton, CA, USA)), pERK (1:1000, rabbit, Cell Signaling (Danvers, MA, USA)), and ERK (1:1000, rabbit, Cell Signaling).

Techniques: Activation Assay, Injection, Western Blot, Phospho-proteomics

Journal: Nutrients

Article Title: Docosahexaenoic Acid Attenuates Visceral Pain by Suppressing Spinal CXCL10/CXCR3/ERK Signaling

doi: 10.3390/nu18071113

Figure Lengend Snippet: CXCL10-induced visceral hypersensitivity and spinal ERK activation are dependent on CXCR3. ( A ) Intrathecal administration of recombinant CXCL10 (100 ng), but not 10 ng, induced visceral hypersensitivity at 1 h and 3 h after injection in naïve mice. The downward arrow indicates the administration time point. **, p < 0.01 vs. PBS, n = 6. ( B ) Pretreatment with the CXCR3 antagonist NBI-74330 significantly attenuated CXCL10 (100 ng)-induced visceral hypersensitivity. *, p < 0.05 vs. vehicle + CXCL10, n = 6. ( C , D ) Western blot analysis and quantification showing that CXCL10-induced ERK phosphorylation in the spinal cord was markedly reduced by NBI-74330 pretreatment. ***, p < 0.001 vs. naïve; ##, p < 0.01 vs. vehicle + CXCL10.

Article Snippet: Membranes were blocked and incubated overnight at 4 °C with primary antibodies against CXCR3 (1:200, rabbit, Boster (Pleasanton, CA, USA)), pERK (1:1000, rabbit, Cell Signaling (Danvers, MA, USA)), and ERK (1:1000, rabbit, Cell Signaling).

Techniques: Activation Assay, Recombinant, Injection, Western Blot, Phospho-proteomics

Journal: Nutrients

Article Title: Docosahexaenoic Acid Attenuates Visceral Pain by Suppressing Spinal CXCL10/CXCR3/ERK Signaling

doi: 10.3390/nu18071113

Figure Lengend Snippet: Intrathecal DHA attenuates TNBS-induced visceral pain and suppresses spinal CXCL10/CXCR3/ERK signaling. ( A ) Intrathecal administration of DHA (10 μg), but not 1 μg, significantly attenuated TNBS-induced visceral hypersensitivity at 1 h and 3 h after injection. *, p < 0.05; **, p < 0.01 vs. TNBS + vehicle, n = 6. ( B , C ) qPCR analysis showing that DHA treatment reduced TNBS-induced upregulation of CXCL10 and CXCR3 mRNA expression in the spinal cord. ***, p < 0.001 vs. naïve; #, p < 0.05 vs. TNBS, n = 6. ( D , E ) Western blot analysis and quantification demonstrated that DHA markedly suppressed TNBS-induced ERK phosphorylation in the spinal cord. **, p < 0.01 vs. naïve; #, p < 0.05 vs. TNBS + vehicle.

Article Snippet: Membranes were blocked and incubated overnight at 4 °C with primary antibodies against CXCR3 (1:200, rabbit, Boster (Pleasanton, CA, USA)), pERK (1:1000, rabbit, Cell Signaling (Danvers, MA, USA)), and ERK (1:1000, rabbit, Cell Signaling).

Techniques: Injection, Expressing, Western Blot, Phospho-proteomics

Journal: Nutrients

Article Title: Docosahexaenoic Acid Attenuates Visceral Pain by Suppressing Spinal CXCL10/CXCR3/ERK Signaling

doi: 10.3390/nu18071113

Figure Lengend Snippet: DHA attenuates LPS-induced CXCL10/CXCR3 expression and ERK activation in neuronal cells. ( A , B ) qPCR analysis showing that LPS markedly increased CXCL10 and CXCR3 mRNA expression in Neuro-2a cells, which was attenuated by DHA pretreatment in a dose-dependent manner. ***, p < 0.001 vs. control; ###, p < 0.001 vs. LPS alone. ( C , D ) Western blot analysis and quantification demonstrating that DHA pretreatment suppressed LPS-induced ERK phosphorylation in Neuro-2a cells. ***, p < 0.001 vs. control; ##, p < 0.01 vs. LPS + vehicle.

Article Snippet: Membranes were blocked and incubated overnight at 4 °C with primary antibodies against CXCR3 (1:200, rabbit, Boster (Pleasanton, CA, USA)), pERK (1:1000, rabbit, Cell Signaling (Danvers, MA, USA)), and ERK (1:1000, rabbit, Cell Signaling).

Techniques: Expressing, Activation Assay, Control, Western Blot, Phospho-proteomics

Journal: Nutrients

Article Title: Docosahexaenoic Acid Attenuates Visceral Pain by Suppressing Spinal CXCL10/CXCR3/ERK Signaling

doi: 10.3390/nu18071113

Figure Lengend Snippet: Mechanism of DHA-mediated visceral pain relief by spinal CXCL10/CXCR3/ERK pathway inhibition. The dot between CXCR3 and phosphorylated ERK represents intermediate signaling molecules in the signaling cascade, which mediate the signal transduction from CXCR3 activation to ERK phosphorylation.

Article Snippet: Membranes were blocked and incubated overnight at 4 °C with primary antibodies against CXCR3 (1:200, rabbit, Boster (Pleasanton, CA, USA)), pERK (1:1000, rabbit, Cell Signaling (Danvers, MA, USA)), and ERK (1:1000, rabbit, Cell Signaling).

Techniques: Inhibition, Transduction, Activation Assay, Phospho-proteomics