Journal: Cancer Science

Article Title: UBE 2 C ‐Mediated CD 147‐ CTLA ‐4 Axis Promotes T Cell Exhaustion and Immunosuppressive Microenvironment in Bladder Cancer

doi: 10.1111/cas.70430

Figure Lengend Snippet: UBE2C knockdown suppresses bladder cancer progression and CTLA‐4 expression. (A) qRT‐PCR analysis of UBE2C expression in T24 cell. (B) Western blot analysis of UBE2C expression in T24 cell. (C) Transwell assay for T24 cell invasion ability. Scale bars: 100 μm. (D) CCK‐8 assay for T24 cell viability. (E) Flow cytometry for T24 cell apoptosis levels. (F) Western blot analysis of Ki‐67 and γ‐H2AX expression in tumor tissue from C57 mice harvested at the experimental endpoint (day 21 postinoculation). (G) Spearman correlation analysis between UBE2C and CTLA‐4 using TCGA‐BLCA database. (H) IHC analysis of CD8 + T cell infiltration in BCa tissue from patients. Scale bars: 100 μm. (I) Flow cytometry analysis of CD3 + CD8 + T cell proportion in tumor tissue. (J) Flow cytometry analysis of CTLA‐4 expression in tumor tissue. *si‐NC versus si‐UBE2C. * p < 0.05, *** p < 0.001.

Article Snippet: One to two weeks after inoculation, the mice were randomly grouped as shNC ( N = 3), shNC + anti‐CTLA‐4 ( N = 3), shUBE2C ( N = 3), and shUBE2C + AAV‐CTLA4 ( N = 3). shNC + anti‐CTLA‐4 mice were treated with anti‐mouse CTLA‐4 (9D9) Antibody (HY‐ P99132 , MCE, USA; 200 μg per mouse) The shUBE2C + AAV‐CTLA‐4 group received CD8 promoter‐specific AAV‐CTLA‐4 treatment (Genomeditech; 30 μg per mouse), and the administration method was peritumoral injection at d7, d10, and d13, while the remaining groups of mice were injected with the same dose of sterile PBS solution.

Techniques: Knockdown, Expressing, Quantitative RT-PCR, Western Blot, Transwell Assay, CCK-8 Assay, Flow Cytometry

Journal: Cancer Science

Article Title: UBE 2 C ‐Mediated CD 147‐ CTLA ‐4 Axis Promotes T Cell Exhaustion and Immunosuppressive Microenvironment in Bladder Cancer

doi: 10.1111/cas.70430

Figure Lengend Snippet: The UBE2C/CTLA‐4 axis drives T cell exhaustion in the bladder cancer microenvironment. (A) CTLA‐4 expression in T cells. (B) UBE2C expression in T24 cells and CTLA‐4 expression in T cells. (C) The expression levels of T cell exhaustion‐related proteins. (D) T cell viability. (E) T cell apoptosis. (F–H) ELISA assay for IFN‐γ (F), TNF‐α (G), and TGF‐β (H) levels in the supernatant. *si‐NC versus si‐UBE2C, # si‐UBE2C versus si‐UBE2C + oe‐CTLA‐4. ## p < 0.01, ### p < 0.001, and *** p < 0.001.

Article Snippet: One to two weeks after inoculation, the mice were randomly grouped as shNC ( N = 3), shNC + anti‐CTLA‐4 ( N = 3), shUBE2C ( N = 3), and shUBE2C + AAV‐CTLA4 ( N = 3). shNC + anti‐CTLA‐4 mice were treated with anti‐mouse CTLA‐4 (9D9) Antibody (HY‐ P99132 , MCE, USA; 200 μg per mouse) The shUBE2C + AAV‐CTLA‐4 group received CD8 promoter‐specific AAV‐CTLA‐4 treatment (Genomeditech; 30 μg per mouse), and the administration method was peritumoral injection at d7, d10, and d13, while the remaining groups of mice were injected with the same dose of sterile PBS solution.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Cancer Science

Article Title: UBE 2 C ‐Mediated CD 147‐ CTLA ‐4 Axis Promotes T Cell Exhaustion and Immunosuppressive Microenvironment in Bladder Cancer

doi: 10.1111/cas.70430

Figure Lengend Snippet: CTLA‐4 overexpression reverses the inhibition of tumor growth mediated by UBE2C knockdown. (A) Mouse diagram. (B) Mouse tumor tissue. (C) Mouse weight. (D) Mouse tumor volume. (E) Mouse tumor weight. *shNC versus shUBE2C or shNC + anti‐CTLA‐4, # shUBE2C versus shUBE2C + AAV‐CTLA‐4. ### p < 0.001, ** p < 0.01 and *** p < 0.001. N = 3.

Article Snippet: One to two weeks after inoculation, the mice were randomly grouped as shNC ( N = 3), shNC + anti‐CTLA‐4 ( N = 3), shUBE2C ( N = 3), and shUBE2C + AAV‐CTLA4 ( N = 3). shNC + anti‐CTLA‐4 mice were treated with anti‐mouse CTLA‐4 (9D9) Antibody (HY‐ P99132 , MCE, USA; 200 μg per mouse) The shUBE2C + AAV‐CTLA‐4 group received CD8 promoter‐specific AAV‐CTLA‐4 treatment (Genomeditech; 30 μg per mouse), and the administration method was peritumoral injection at d7, d10, and d13, while the remaining groups of mice were injected with the same dose of sterile PBS solution.

Techniques: Over Expression, Inhibition, Knockdown

Journal: Cancer Science

Article Title: UBE 2 C ‐Mediated CD 147‐ CTLA ‐4 Axis Promotes T Cell Exhaustion and Immunosuppressive Microenvironment in Bladder Cancer

doi: 10.1111/cas.70430

Figure Lengend Snippet: Knocking down UBE2C inhibits bladder cancer growth by alleviating CTLA‐4‐mediated immune suppression. (A, B) Immunohistochemistry (A) and Western blot (B) analysis of UBE2C and CTLA‐4 expression in tumor tissues. Scale bars: 100 μm. (C) T cell exhaustion protein expression in tumor tissues. (D–F) ELISA analysis of IFN‐γ (D), TNF‐α (E), and TGF‐β (F) levels in mouse blood. *shNC versus shUBE2C or shNC + anti‐CTLA‐4, # shUBE2C versus shUBE2C + AAV‐CTL A‐4. ### p < 0.001 and *** p < 0.001. N = 3.

Article Snippet: One to two weeks after inoculation, the mice were randomly grouped as shNC ( N = 3), shNC + anti‐CTLA‐4 ( N = 3), shUBE2C ( N = 3), and shUBE2C + AAV‐CTLA4 ( N = 3). shNC + anti‐CTLA‐4 mice were treated with anti‐mouse CTLA‐4 (9D9) Antibody (HY‐ P99132 , MCE, USA; 200 μg per mouse) The shUBE2C + AAV‐CTLA‐4 group received CD8 promoter‐specific AAV‐CTLA‐4 treatment (Genomeditech; 30 μg per mouse), and the administration method was peritumoral injection at d7, d10, and d13, while the remaining groups of mice were injected with the same dose of sterile PBS solution.

Techniques: Immunohistochemistry, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Cancer Science

Article Title: UBE 2 C ‐Mediated CD 147‐ CTLA ‐4 Axis Promotes T Cell Exhaustion and Immunosuppressive Microenvironment in Bladder Cancer

doi: 10.1111/cas.70430

Figure Lengend Snippet: UBE2C promotes the expression of CTLA‐4 by regulating CD147 and induces T cell exhaustion. (A, B) CD147 expression in T24 cells (A) and mouse tumor tissue (B). (C) CD147 expression in T24 cells. (D, E) T cell apoptosis. (F) T cell proliferation activity. (G) UBE2C expression in T24 cells and CTLA‐4 expression in T cells. (H) T cell exhaustion proteins. (I–K) ELISA assay for IFN‐γ (I), TNF‐α (J), and TGF‐β (K) levels in cell culture supernatants. (L) ELISA detection of soluble CD147 (sCD147) levels in the conditioned medium. (M) Western blot analysis of exosomal markers CD63 and CD9 in isolated extracellular vesicles. (N) Western blot analysis of CD147 expression in tumor‐derived exosomes. *si‐NC versus si‐UBE2C or si‐CD147, # si‐UBE2C versus si‐UBE2C + oe‐CD147. ### p < 0.001 and *** p < 0.001.

Article Snippet: One to two weeks after inoculation, the mice were randomly grouped as shNC ( N = 3), shNC + anti‐CTLA‐4 ( N = 3), shUBE2C ( N = 3), and shUBE2C + AAV‐CTLA4 ( N = 3). shNC + anti‐CTLA‐4 mice were treated with anti‐mouse CTLA‐4 (9D9) Antibody (HY‐ P99132 , MCE, USA; 200 μg per mouse) The shUBE2C + AAV‐CTLA‐4 group received CD8 promoter‐specific AAV‐CTLA‐4 treatment (Genomeditech; 30 μg per mouse), and the administration method was peritumoral injection at d7, d10, and d13, while the remaining groups of mice were injected with the same dose of sterile PBS solution.

Techniques: Expressing, Activity Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Western Blot, Isolation, Derivative Assay

Journal: Cancer Science

Article Title: UBE 2 C ‐Mediated CD 147‐ CTLA ‐4 Axis Promotes T Cell Exhaustion and Immunosuppressive Microenvironment in Bladder Cancer

doi: 10.1111/cas.70430

Figure Lengend Snippet: UBE2C upregulates cellular CD147 and enhances its release into the tumor microenvironment both as soluble CD147 (sCD147) and packaged within tumor‐derived exosomes. These CD147‐bearing signals induce T‐cell exhaustion, marked by elevated CTLA‐4 expression and reduced secretion of effector cytokines (IFN‐γ, TNF‐α, TGF‐β), ultimately diminishing CD8 + T cell abundance and antitumor immunity.

Article Snippet: One to two weeks after inoculation, the mice were randomly grouped as shNC ( N = 3), shNC + anti‐CTLA‐4 ( N = 3), shUBE2C ( N = 3), and shUBE2C + AAV‐CTLA4 ( N = 3). shNC + anti‐CTLA‐4 mice were treated with anti‐mouse CTLA‐4 (9D9) Antibody (HY‐ P99132 , MCE, USA; 200 μg per mouse) The shUBE2C + AAV‐CTLA‐4 group received CD8 promoter‐specific AAV‐CTLA‐4 treatment (Genomeditech; 30 μg per mouse), and the administration method was peritumoral injection at d7, d10, and d13, while the remaining groups of mice were injected with the same dose of sterile PBS solution.

Techniques: Derivative Assay, Expressing

Journal: bioRxiv

Article Title: A class act: HDAC1- Malat1 regulates MDSC apoptosis and cell cycling to decrease suppression of T cells

doi: 10.64898/2026.03.23.713743

Figure Lengend Snippet: (A) UMAP visualization of major cell clusters identified from single cell RNA sequencing (scRNA-seq) of NT2.5-LM breast-to-lung metastatic tumors after 3 weeks of treatment with: vehicle (V), Entinostat (E), anti-CTLA-4 + anti-PD-1 immune checkpoint inhibitors (PC), and Entinostat + anti-CTLA-4 + anti-PD-1 combination (EPC). Subclusters of MDSCs identified: G-MDSCs and M-MDSCs. (B) Iterative logistic regression analyses conducted on the major MDSC cluster comparing the Vehicle (V) and Entinostat + ICIs combination treatment (EPC). (C) Expression of Malat1 / MALAT1 in G- and M-MDSCs isolated from lung metastases of NT2.5-LM mice treated with vehicle vs. Entinostat for 3 weeks (left, n=2 from 5 pooled mice per treatment group), J774M murine MDSC-like cell line treated with Entinostat for 24 hours (middle, n=3), and human PBMC-derived MDSCs (hMDSCs) treated with Entinostat for 24 hours (right, n=3). (D) Median Fluorescence Intensity expression of Malat1 in CD45 + CD11b + MHC-II - F4/80 - Ly6G - Ly6C hi cells from lung metastases in 4T1 mice, treated with vehicle (V), Entinostat (E), and Entinostat + ICIs (EPC) for 3 weeks. (E) Expression of Malat1 in J774M cell line treated with various concentrations of various HDAC inhibitors, with corresponding targeted class and specific HDACs. One-way ANOVA for (C, E: all statistically significant with p<0.05, unless indicated as non-significant (ns)), Kruskal-Wallis test with Dunn’s correction for (D). * p<0.05, *** p< 0.001, **** p<0.0001. ENT = Entinostat; PAN = Panobinostat; BEL = Belinostat; VOR = Vorinostat; TSA = Trichostatin A; DOM = Domatinostat; CHL = Chlopynostat; PYR = Pyroxamide; SCA = Santacruzamate A; RGF = RGFP966; TMP = TMP269; SIS = SIS17.

Article Snippet: Immune checkpoint inhibitors (ICIs) used in this study: anti-CTLA-4 (BioXCell cat. #BE0131) and anti-PD-1 (BioXCell cat. #BE0146).

Techniques: Single Cell, RNA Sequencing, Expressing, Isolation, Derivative Assay, Fluorescence

Journal: Cell Reports Medicine

Article Title: Neoadjuvant Fc-enhanced anti-CTLA-4 targets Tregs to augment androgen deprivation in high-risk prostate cancer: A randomized phase I trial

doi: 10.1016/j.xcrm.2026.102638

Figure Lengend Snippet: Effect of ADT + Fc-enhanced anti-CTLA-4 on DCs (A) Violin plot of myeloid cluster frequencies as percentage of myeloid cells stratified by patient’s 2-year PSA recurrence status. No recurrence, n = 7; recurrence, n = 4. Two-tailed Student’s t test was utilized to test statistical significance. For this and all violin plots to follow, solid lines denote group medians, while dashed lines denote quartiles. (B) Kaplan-Meier curve representing time-to-PSA recurrence in patients stratified by dendritic cell frequency; 95% confidence intervals shown in the shaded areas. There was one recurrence event in DC-high patients and three recurrence events in DC-low patients. Log rank test was performed to evaluate statistical significance. (C) Schematic of pre-clinical validation experiment in the MycCaP model. (D) Percent change in tumor volume relative to baseline volume prior to ADT stratified by treatment group. Average values are represented by solid lines and standard deviations are represented by shaded areas. (E) Kaplan-Meier curve representing survival of mice shown in (D). Log rank (Mantel-Cox) test was used to evaluate statistical significance. (F) Left: supervised UMAP of live CD45 + cells from MycCaP tumors harvested following therapy as denoted in (D and E). Semi-supervised clustering was performed using FlowSOM and clusters were manually annotated according to expression of lineage-defining markers. Right: heatmap of lineage-defining markers across all clusters. Data represent geometric MFI for each marker normalized to the minimum and maximum values across all clusters. (G and H) (G) Frequency of TI-Tregs and (H) frequency of cDCs as percentage of all live CD45 + cells, stratified by treatment group. (I) Left: PaCMAP representing FlowSOM-derived cDC clusters. Clusters were manually annotated according to expression of canonical marker proteins. Right: pseudocolor representation of cDC phenotypes in PaCMAP space stratified by treatment group. (J) Violin plot representing frequency of PD-L2 + DCs as percentage of all DCs stratified by treatment group. (K) Violin plot representing frequency of CD40 + DCs as percentage of all DCs stratified by treatment group. Welch’s t test was used to assess statistical significance. Murine data shown are n = 8 mice per group and representative of two independent experiments each for survival and immune profiling studies.

Article Snippet: RecombiMab anti-Mouse CTLA-4 9D9-mIgG2a , BioXCell , 9D9-CP007.

Techniques: Two Tailed Test, Biomarker Discovery, Expressing, Marker, Derivative Assay

Journal: Cell Reports Medicine

Article Title: Neoadjuvant Fc-enhanced anti-CTLA-4 targets Tregs to augment androgen deprivation in high-risk prostate cancer: A randomized phase I trial

doi: 10.1016/j.xcrm.2026.102638

Figure Lengend Snippet: Effect of ADT + Fc-enhanced anti-CTLA-4 on DCs (A) Violin plot of myeloid cluster frequencies as percentage of myeloid cells stratified by patient’s 2-year PSA recurrence status. No recurrence, n = 7; recurrence, n = 4. Two-tailed Student’s t test was utilized to test statistical significance. For this and all violin plots to follow, solid lines denote group medians, while dashed lines denote quartiles. (B) Kaplan-Meier curve representing time-to-PSA recurrence in patients stratified by dendritic cell frequency; 95% confidence intervals shown in the shaded areas. There was one recurrence event in DC-high patients and three recurrence events in DC-low patients. Log rank test was performed to evaluate statistical significance. (C) Schematic of pre-clinical validation experiment in the MycCaP model. (D) Percent change in tumor volume relative to baseline volume prior to ADT stratified by treatment group. Average values are represented by solid lines and standard deviations are represented by shaded areas. (E) Kaplan-Meier curve representing survival of mice shown in (D). Log rank (Mantel-Cox) test was used to evaluate statistical significance. (F) Left: supervised UMAP of live CD45 + cells from MycCaP tumors harvested following therapy as denoted in (D and E). Semi-supervised clustering was performed using FlowSOM and clusters were manually annotated according to expression of lineage-defining markers. Right: heatmap of lineage-defining markers across all clusters. Data represent geometric MFI for each marker normalized to the minimum and maximum values across all clusters. (G and H) (G) Frequency of TI-Tregs and (H) frequency of cDCs as percentage of all live CD45 + cells, stratified by treatment group. (I) Left: PaCMAP representing FlowSOM-derived cDC clusters. Clusters were manually annotated according to expression of canonical marker proteins. Right: pseudocolor representation of cDC phenotypes in PaCMAP space stratified by treatment group. (J) Violin plot representing frequency of PD-L2 + DCs as percentage of all DCs stratified by treatment group. (K) Violin plot representing frequency of CD40 + DCs as percentage of all DCs stratified by treatment group. Welch’s t test was used to assess statistical significance. Murine data shown are n = 8 mice per group and representative of two independent experiments each for survival and immune profiling studies.

Article Snippet: RecombiMab anti-Mouse CTLA-4 9D9-mIgG2a-LALAPG , BioXCell , 9D9-CP008.

Techniques: Two Tailed Test, Biomarker Discovery, Expressing, Marker, Derivative Assay