Journal: Advanced Science

Article Title: Oral Astilbin Nanogels Modulate Senescence‐Associated CD4 + T Cells and Promote T reg Induction in Aged Mice with Colitis

doi: 10.1002/advs.78123

Figure Lengend Snippet: Ast attenuates senescence‐associated features and restores the immunoregulatory function of CD4 + T cells. (A) Correlation between PDCD1 expression, which encodes PD‐1, and the age at diagnosis in patients with colitis ( n = 106). (B) Comparison of PDCD1 expression levels between young (< 60 years old) and aged (≥ 60 years old) colitis patients. (C) Experimental flowchart for DSS‐induced colitis in young and aged mice. (D) Curve chart analysis of DAI ( n = 6). (E) Representative images of colons and corresponding quantification of colon length ( n = 6). (F) Quantitative analysis of CD4 + PD‐1 + T cells and CD4 + SA‐β‐gal + T cells in colonic lamina propria cells and representative FC plots ( n = 6). (G) In vitro effects of different concentrations of Ast on PD‐1 expression in colonic CD4 + T cells isolated from young and aged mice ( n = 5). (H) In vitro proliferation of CD4 + T cells isolated from aged mice following Ast treatment ( n = 3). (I) In vitro differentiation of CD4 + T cells isolated from aged mice into T regs following Ast treatment ( n = 3). Data are presented as mean ± SD and analyzed by two‐tailed unpaired Student's t test with Welch's correction (B), two‐tailed unpaired Student's t test (H, I), Two‐way ANOVA with Tukey's post‐test (D), or One‐way ANOVA with Tukey's post‐test (E–G). ns represents not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: Purified blocking anti‐mouse PD‐1 (Clone RMP1‐14), agonistic anti‐mouse PD‐1 (Clone RMP1‐30), anti‐mouse CD3 (Clone 17A2), and anti‐mouse CD28 (Clone 37.51) were purchased from BioXcell (USA).

Techniques: Expressing, Biomarker Discovery, Comparison, Analysis, In Vitro, Isolation, Two Tailed Test

Journal: Advanced Science

Article Title: Oral Astilbin Nanogels Modulate Senescence‐Associated CD4 + T Cells and Promote T reg Induction in Aged Mice with Colitis

doi: 10.1002/advs.78123

Figure Lengend Snippet: High‐dose free Ast alleviates colitis in young and aged mice by attenuating senescence‐associated immune dysfunction. (A) Experimental flowchart of young and aged colitis mice treated with free Ast. (B) Curve chart variations of murine body weight across time, normalized to the day 0 body weight percentage ( n = 6). (C) Curve chart analysis of DAI ( n = 6). (D) Representative images of colons and corresponding quantification of colon length in young and aged mice ( n = 6). Representative FC plots and quantitative analysis of (E) CD4 + PD‐1 + T cells, (F) CD4 + SA‐β‐gal + T cells, and (G) CD25 + Foxp3 + CD4 T cells in colonic lamina propria cells ( n = 6). (H) Heatmap of differentially expressed genes in colon samples ( n = 6). GSEA of the (I) TGF‐beta signaling pathway and (J) AGE‐RAGE signaling pathway. Data are presented as mean ± SD and analyzed by One‐way ANOVA with Tukey's post‐test (D‐G) or Two‐way ANOVA with Tukey's post‐test (B, C). ns represents not significant, * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Purified blocking anti‐mouse PD‐1 (Clone RMP1‐14), agonistic anti‐mouse PD‐1 (Clone RMP1‐30), anti‐mouse CD3 (Clone 17A2), and anti‐mouse CD28 (Clone 37.51) were purchased from BioXcell (USA).

Techniques: Analysis

Journal: Advanced Science

Article Title: Oral Astilbin Nanogels Modulate Senescence‐Associated CD4 + T Cells and Promote T reg Induction in Aged Mice with Colitis

doi: 10.1002/advs.78123

Figure Lengend Snippet: Mechanistic validation of Ast‐mediated regulation of senescence‐associated CD4 + T cells in aged colitis. (A) Experimental flowchart of NG treatment in aged colitis mice ( n = 6). (B) Representative fecal samples following occult blood testing. (C) Curve chart analysis of DAI ( n = 6). (D) Representative images of colons and corresponding quantification of colon length ( n = 6). (E) Quantification of PD‐1 + CD4 T cells in colonic lamina propria cells ( n = 6). (F) Representative FC plots and quantitative analysis of CD4 + CD27 − CD28 − T cells in colonic lamina propria cells ( n = 6). (G) Schematic illustration of the isolation and sorting strategy for senescence‐associated (CD28 − CD27 − ) and non‐senescence‐associated (remaining) CD4 + T cells. Created with BioRender. (H) MFI of PD‐1 expression on senescence‐associated and non‐senescence‐associated CD4 + T cells after treatment with PBS or Ast for 2 days ( n = 3). (I) Cell viability of senescence‐associated and non‐senescence‐associated CD4 + T cells ( n = 3). (J) Foxp3 + T reg differentiation capacity of senescence‐associated and non‐senescence‐associated CD4 + T cells in the presence of PBS or Ast for 2 days ( n = 3). (K) Experimental groups for therapeutic evaluation in aged colitis mice. (L) Curve chart analysis of DAI ( n = 6). (M) Quantification of colon length ( n = 6). Quantitative analysis of (N) CD4 + PD‐1 + T cells and (O) CD4 + CD27 − CD28 − T cells in colonic lamina propria cells ( n = 6). (P) Schematic illustration of CD4 + T cells cultured in the presence of plate‐bound agonistic aPD‐1 or Ast. Created with BioRender. (Q) Quantitative analysis of CD4 + CD27 − CD28 − T cells following treatment with plate‐bound agonistic aPD‐1 or Ast ( n = 3). Data are presented as mean ± SD and analyzed by Two‐way ANOVA with Tukey's post‐test (C, L) or One‐way ANOVA with Tukey's post‐test (D‐F, H‐J, M‐O, Q). ns represents not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: Purified blocking anti‐mouse PD‐1 (Clone RMP1‐14), agonistic anti‐mouse PD‐1 (Clone RMP1‐30), anti‐mouse CD3 (Clone 17A2), and anti‐mouse CD28 (Clone 37.51) were purchased from BioXcell (USA).

Techniques: Biomarker Discovery, Fecal/Stool Samples, Analysis, Isolation, Expressing, Cell Culture, FACS

Journal: bioRxiv

Article Title: Neutrophils orchestrate pre-metastatic immunosuppressive bone marrow remodeling during prostate cancer metastasis

doi: 10.64898/2026.10.08.757800

Figure Lengend Snippet: (a) Experimental design for testing whether conditioned-medium (CM)-induced BM preconditioning promotes bone metastasis and whether PPP prevents this effect. Mice received matched blank medium (Control), wild-type TRAMP-C1 CM or CM plus PPP before intracardiac TRAMP-C1 injection. (b) Experimental design for testing whether recombinant IGF1-induced BM preconditioning promotes bone metastasis and whether PPP prevents this effect. Control mice received matched IGF1 and PPP vehicles. (c, d) Bone-metastasis-free curves and longitudinal bone bioluminescence imaging (BLI) signals after CM (c) or IGF1 (d) preconditioning, with or without PPP. For longitudinal BLI analyses, the area under the curve (AUC) was calculated for each mouse from ln(signal + 1)-transformed serial BLI measurements, and per-mouse AUC values were compared between groups; n = 10 mice per group. P 1 denotes Control versus CM in c and Control versus IGF1 in d; P 2 denotes CM versus CM plus PPP in c and IGF1 versus IGF1 plus PPP in d. (e) Experimental design using neutrophil-specific Igf1r conditional-knockout (cKO) mice and littermate controls to evaluate IGF1-induced BM preconditioning and subsequent bone metastasis. (f) Bone-metastasis-free curves and longitudinal bone BLI signals in control and cKO mice. BLI signals were natural-log transformed as ln(signal + 1); n = 10 mice per group. (g) Experimental design of the metastatic model evaluating Control, anti-IGF1R, anti-PD-1 or combined anti-IGF1R plus anti-PD-1 treatment. Control mice received matched saline. (h) Bone-metastasis-free curves and longitudinal bone BLI signals across treatment groups. BLI signals were natural-log transformed as ln(signal + 1); n = 10 mice per group. P values for the prespecified pairwise comparisons are shown in the panel: P 1, P 2 and P 3 denote Control versus Combination, Saline versus anti-IGF1R and Saline versus anti-PD-1, respectively. (i) Representative haematoxylin-and-eosin-stained bone sections from control, anti-IGF1R-, anti-PD-1- and combination-treated mice. Dashed lines outline tumor regions (T); B denotes bone. Scale bar, 400 µm. (j) Quantification of tumor area in the indicated treatment groups; each dot represents one mouse. (k) Frequencies of CCR3-positive neutrophils, IFNG-positive and PD1-positive CD8 T cells, and Tregs, together with BM IL-1β concentrations, across treatment groups; n = 3 mice per group. (l) Flowchart of the investigator-initiated trial of teprotumumab plus tislelizumab in metastatic castration-resistant prostate cancer; seven patients were enrolled and received treatment; one subsequently withdrew from the study. (m) Best percentage change in serum prostate-specific antigen (PSA) from baseline. (n) PSA progression-free survival from the first dose. (o) Representative axial PET/CT images before treatment and at day 84. (p) Best percentage change in serum alkaline phosphatase (ALP) from baseline. (q) Patient-reported pain outcomes at days 28, 56 and 84 after the first dose, categorized as improved, stable or worse relative to the immediately preceding assessment (day 28 versus baseline, day 56 versus day 28 and day 84 versus day 56); n = 6 evaluable patients at each time point. Unless otherwise indicated, bar plots with error bars show mean ± s.d. Bone-metastasis-free curves were compared using log-rank tests. Longitudinal BLI data were compared using a two-sided Welch’s t-test. Pairwise cellular or histological comparisons were analyzed using two-sided Student’s t-tests, with Benjamini-Hochberg adjustment where multiple comparisons were performed. Exact P values are shown in the panels.

Article Snippet: For the therapeutic bone-metastasis model, TRAMP-C1-LUC cells were introduced without prior CM or IGF1 preconditioning, and treatment was initiated 1 day after intracardiac tumor-cell injection, anti-IGF1R antibody ganitumab (MCE, HY-P99294, CAS 905703-97-1; 1 mg per mouse loading dose followed by 300 μg per mouse maintenance doses, administered intraperitoneally twice weekly), anti-PD-1 antibody (Bio X Cell, BE0146; 200 μg per dose, intraperitoneally, three doses at 3-day intervals) or the combination according to the treatment schedule shown in .

Techniques: Control, Injection, Recombinant, Imaging, Transformation Assay, Gene Knockout, Saline, Staining, Positron Emission Tomography-Computed Tomography

Journal: bioRxiv

Article Title: Neutrophils orchestrate pre-metastatic immunosuppressive bone marrow remodeling during prostate cancer metastasis

doi: 10.64898/2026.10.08.757800

Figure Lengend Snippet: (a) Heatmap of genes differentially expressed in control and IGF1-treated mouse BM neutrophils; values are scaled expressions. Two mice were analysed per group, with each mouse serving as an independent biological replicate. (b) Gene-set enrichment analysis of the TAN signature in IGF1-treated versus control neutrophils using the RNA-seq data shown in a (n = 2 mice per group). (c) Representative immunofluorescence images of peripheral-blood and BM neutrophils from control- or IGF1-treated mice stained for DAPI, CD274, Ly6G and CCR3. Scale bars, 10 µm. (d) Frequencies of CCR3-positive and CD274-positive neutrophils in peripheral blood and BM; n = 3 mice per group. (e) Transcription-factor motifs enriched in accessible chromatin after IGF1 stimulation. n = 2 biologically independent neutrophil preparations per group. (f) Intersection of IGF1-upregulated genes (RNA-seq), IGF1-regulated transcription factors (ATAC-seq) and TAN-upregulated genes (scRNA-seq), identifying KLF4 as a candidate regulator. (g) KLF4 MFI in neutrophils after IGF1 stimulation with or without MAPK/ERK inhibition; n = 3 biologically independent samples per group. (h) ChIP-qPCR analysis of c-Myc occupancy at the KLF4 promoter after IGF1 stimulation; normal IgG served as the negative control. n = 3 biologically independent samples per group. (i) Experimental design for IGF1-induced BM preconditioning in mice receiving matched vehicle (Control), IGF1 or IGF1 plus the IGF1R inhibitor picropodophyllin (PPP). (j) CCR3 and ICAM1 MFIs in BM neutrophils after the indicated treatments; n = 6 mice per group. (k) Representative BM immunofluorescence images stained for DAPI, CCR3 and Ly6G. Scale bars, 50 µm in the main images and 5 µm in the insets. (l) Experimental design of the CM-induced BM-preconditioning model using matched blank medium (Control), wild-type TRAMP-C1 conditioned-medium (WT CM), Igf1-knockdown CM (Igf1-KD CM) or wild-type CM combined with an IGF1R-blocking antibody. (m) Frequencies of CCR3-positive neutrophils, PD-1-positive CD8 T cells and Tregs after the indicated CM treatments; n = 3 mice per group. (n) Experimental design for analysis of IGF1-induced BM remodeling in neutrophil-specific Igf1r conditional-knockout (cKO) mice and littermate controls. (o) CD274, CCR3 and ICAM1 MFIs in neutrophils, intermediate-monocyte frequency and CD8Temra frequency in control and cKO mice after IGF1 treatment; n = 3 mice per group. (p) Design of the mouse BM scRNA-seq experiment comprising saline-control, IGF1-induced HMP-like, and IGF1-preconditioned TRAMP-C1 metastatic groups. For each experimental group, BM cells from three mice were pooled to generate one scRNA-seq sample. (q) UMAP of all mouse BM cells, colored by major cell type. (r) UMAP of mouse neutrophil states with group-specific embeddings. (s) Human-derived TAN signature scores across mouse neutrophil states. (t) Differential abundance of mouse neutrophil states across experimental groups. Color indicates the log odds ratio, and “+” or “-” denotes significant enrichment or depletion, respectively. (u) UMAP of mouse BM CD8 T cell subtypes with group-specific embeddings. (v) Differential abundance of CD8 T cell subtypes across experimental groups. (w) IL-1β-induced signature scores in CD8Temra cells across saline-control, IGF1-induced HMP and TRAMP-C1 metastatic groups. Unless otherwise indicated, bar plots with error bars show mean ± s.d. Statistical comparisons in panels d, g, h, j, m and o were performed using two-sided Student’s t-tests. P values were adjusted using the Benjamini–Hochberg method where multiple comparisons were performed. For the GSEA in panel b, nominal P values and normalized enrichment scores are shown.

Article Snippet: For the therapeutic bone-metastasis model, TRAMP-C1-LUC cells were introduced without prior CM or IGF1 preconditioning, and treatment was initiated 1 day after intracardiac tumor-cell injection, anti-IGF1R antibody ganitumab (MCE, HY-P99294, CAS 905703-97-1; 1 mg per mouse loading dose followed by 300 μg per mouse maintenance doses, administered intraperitoneally twice weekly), anti-PD-1 antibody (Bio X Cell, BE0146; 200 μg per dose, intraperitoneally, three doses at 3-day intervals) or the combination according to the treatment schedule shown in .

Techniques: Control, Analysis, RNA Sequencing, Immunofluorescence, Staining, Inhibition, ChIP-qPCR, Negative Control, Knockdown, Blocking Assay, Gene Knockout, Saline, Derivative Assay, Oligomer Restriction