multiplex immunofluorescence  (Vectra Laboratories)

Journal: Signal Transduction and Targeted Therapy

Article Title: Single-cell imaging analysis, therapeutic modeling and a Phase Ib trial validate BCL-2 as a target across heterogeneous castration-resistant prostate cancer

doi: 10.1038/s41392-026-02700-w

Figure Lengend Snippet: qmIF analysis reveals predominant AR + BCL-2 - cells in untreated primary PCa and markedly increased (AR + or AR - ) BCL-2 + cells in CRPC. a In benign prostatic glands (HPCa14N), BCL-2 + cells are mainly in the basal cell layer whereas AR + cells in luminal layer. Cytokeratin (CK) staining was used to mark epithelial compartment. Note both AR and BCL-2 were also expressed in stromal cells (scale bar, 50 μm). Magnified images of individual stains from the boxed region in the whole-mount (WM) image (top) were presented below (scale bar, 20 μm for all 4 lower panels). b Primary PCa is characterized by significantly increased AR + BCL-2 - cells. Shown above are two WM images of HPCa31T (scale bars, 800 μm) and down below zoom-in images of individual or merged markers (scale bars, 80 μm). c , d Increased cellular heterogeneity and markedly expanded (AR + or AR - ) BCL-2 + cell population in CRPC. c WM low-magnification image showing AR/BCL-2 expression (scale bar, 300 μm). d Zoom-in images of the boxed area in c showing individual markers (scale bar, 50 μm). e Relationship between (CK + ) AR-expressing and/or BCL-2-expressing cells in benign tissue (top), primary tumor (middle) and CRPC (bottom). Regression line, and Pearson R and P values are indicated. f Box plots summarizing the relative % of 4 PCa cell subtypes in WM images analyzed in benign tissues, primary tumors and CRPC. Each dot in the box plots represents a CK + ROI. P values were determined by repeated measures two-way ANOVA with Bonferroni multiple comparison test

Article Snippet: To confirm and extend these findings, we employed Vectra-based quantitative multiplex immunofluorescence (qmIF) to assess PCa cells expressing BCL-2 and/or AR in regular FFPE (formalin-fixed and paraffin-embedded) sections as well as TMA (tissue microarray) and whole-mount (WM) sections from benign prostate ( n = 123), treatment-naïve primary PCa ( n = 125) and treatment-failed CRPC ( n = 25) (Fig. ; supplementary Figs. – , supplementary Table ).

Techniques: Staining, Expressing, Comparison

Journal: Signal Transduction and Targeted Therapy

Article Title: Single-cell imaging analysis, therapeutic modeling and a Phase Ib trial validate BCL-2 as a target across heterogeneous castration-resistant prostate cancer

doi: 10.1038/s41392-026-02700-w

Figure Lengend Snippet: Vectra and IMC analysis reveals castration-induced dynamic changes in AR +/- BCL-2 +/- cell types in 4 xenograft CRPC models. a – d tSNE plots of IMC-derived single-cell imaging data. Shown are the individual and merged tSNE plots from LNCaP-AD/AI ( a ), LAPC9-AD/AI ( b ), LAPC4-AD/AI ( c ), and VCaP-AD/AI ( d ) xenografts. Columns represent merged AD and AI cell clusters (first column), individual AD and AI cell populations and contours (second and third columns), and AR and BCL-2 expression overlaid on tSNE maps (fourth and fifth columns). Blue and red contours indicate AD and AI cell subpopulations, respectively. e LAPC4-AD tumors are populated mostly by AR + BCL-2 - PCa cells. Shown on top are qmIF WM images (scale bar, 800 μm) and at the bottom zoom-in images (scale bar, 80 μm for all panels). f LAPC4-AI (1° CRPC) tumors are populated by AR cyto BCL-2 + PCa cells. Shown on top are qmIF WM images (scale bar, 800 μm) and at the bottom zoom-in images illustrating cytoplasmic AR + LAPC4-AI cells with upregulated BCL-2 (scale bar, 80 μm for all panels). g IMC images of AR and BCL-2 in LAPC4-AD and LAPC4-AI tumors (scale bar, 200 μm) with representative zoom-in images of AR and BCL-2 shown below (scale bar, 100 μm for all panels)

Article Snippet: To confirm and extend these findings, we employed Vectra-based quantitative multiplex immunofluorescence (qmIF) to assess PCa cells expressing BCL-2 and/or AR in regular FFPE (formalin-fixed and paraffin-embedded) sections as well as TMA (tissue microarray) and whole-mount (WM) sections from benign prostate ( n = 123), treatment-naïve primary PCa ( n = 125) and treatment-failed CRPC ( n = 25) (Fig. ; supplementary Figs. – , supplementary Table ).

Techniques: Derivative Assay, Single Cell, Imaging, Expressing