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: Single-cell multi-omic interrogation of mechanisms underlying TI-Treg modulation and clinical response to anti-CTLA4-NF (A) Schematic of immune correlates performed. (B) Summary of data collection and QC passing at a per-patient level for CyTOF, scRNA-seq, and immunofluorescence datasets. Patients are additionally annotated according to presence or absence of PSA recurrence within 2 years post-surgery. (C) UMAP plot of single-cell RNA-seq data from all patients, clustered using the PICSES analytical pipeline. (D) Bubble plot of representative gene expression for each cluster identified in (C). (E) Supervised UMAP plot of CyTOF data from all patients, clustered using FlowSOM. (F) Heatmap of lineage-defining protein expression across all clusters. Data represent geometric MFI for each marker normalized to the minimum and maximum values across all clusters. (G) Heatmap representing Log 2 fold change of scRNA-seq-defined cluster frequencies in indicated treatment group relative to the untreated group. (H) Heatmap representing Log 2 fold change of CyTOF-defined immune cluster frequencies in indicated treatment group relative to the untreated group. (I) UMAP plot representing subclustering of myeloid cells in the scRNA-seq dataset utilizing the PISCES pipeline. (J) Visualization of representative myeloid-lineage-defining gene expression overlayed on the parent NeoRED tumor myeloid scRNA-seq UMAP plot to validate cluster annotations. (K) Bubble plot validating expression of additional myeloid cluster defining genes. (L) Visualization of VIPER-inferred protein activity of select Fcγ receptors across myeloid clusters in UMAP space. (M) Violin plot of FCGR3A /CD16a inferred protein activity on a per cell level stratified by myeloid cluster. (N) Correlation between Treg frequency and average inferred protein activity of FCGR3A /CD16a across all myeloid cells on a per patient basis, stratified by treatment group. ADT, n = 8; ADT + anti-CTLA4-NF, n = 11. (O) Correlation between Treg frequency and average inferred protein activity of FCGR3A /CD16a across TREM2 + TAM cluster cells on a per patient basis, stratified by treatment group. Pearson correlations were utilized to evaluate statistical significance of correlations. (P) Bubble plot representing relative expression of phenotypic markers in Tregs from ADT + anti-CTLA4-NF group versus Tregs in the ADT only group, as calculated by log 2 transformed ratio of average geometric MI of each marker on Tregs from ADT + anti-CTLA4-NF versus ADT groups. Bubble size represents relative gMI, while color intensity represents percent expression of indicated markers across all Tregs in the dataset. Red color indicates higher relative gMI in Tregs from patients treated with ADT + anti-CTLA4-NF, while blue color indicates higher relative gMI in patients treated with ADT only. (Q) Violin plot representing frequency of Tregs at time of surgery in the scRNA-seq dataset stratified by 2-year PSA recurrence status. No recurrence, n = 7; recurrence, n = 4. Two-tailed Welch’s t test was performed to evaluate statistical significance. Solid lines denote group median, and dashed lines denote quartiles. (R) Kaplan-Meier curve representing time-to-PSA recurrence in ADT + anti-CTLA4-NF-treated patients stratified by Treg frequency in the scRNA-seq data; 95% confidence intervals shown in the shaded areas. There were three recurrence events in Treg-high patients and one recurrence event in Treg-low patients. Log rank test was performed to evaluate statistical significance. Unless otherwise stated, two-tailed Welch’s t test was used to assess statistical significance.

Article Snippet: Anti-Human CD16 (3G8) 148ND , Standard BioTools , Cat# 3148004B; RRID: AB_10900638.

Techniques: Single Cell, Immunofluorescence, RNA Sequencing, Gene Expression, Expressing, Marker, Activity Assay, Transformation Assay, Two Tailed Test