Fig. 1: Establishment of a novel androgen dependent, patient-derived xenograft from an early, treatment-naïve prostate cancer metastasis.
From: Patient-derived xenografts and organoids model therapy response in prostate cancer
a Scheme of clinical history and patient-derived samples: primary tumor TUR-P (T1) and penile metastasis needle biopsies used to establish the PDX model (PNPCa) and subsequent passages. Created with BioRender.com. b Histological morphology of primary TURP tumor, penile metastasis (PN met) from PCa and the PDX passages 1 and 6 (PDX1, PDX6) derived from the metastasis needle biopsy implantation (PNPCa), as assessed by Hematoxylin and Eosin staining (H&E). Scale bars 20 μm. Top to bottom panels: PSA protein expression. Scale bars 20 μm. Expression of AR (green), CK5 (red) assessed by immunofluorescence, DAPI (blue) marks the nuclei. Scale bars 50 μm. Expression of NKX3.1 (green), CK8 (red) assessed by immunofluorescence. Scale bars 50 μm. c Flow cytometry analysis of epithelial and prostate-specific marker expression in PNPCa PDX tissue. FcR-blocked PNPCa cells were stained with antibodies against CD44, E-Cadherin, PSMA, CD49f, CD36, and CD146. d PDX tumor growth progression in time. Groups; 1. Intact tumors (collected at max size, N = 2 independent animals), 2. Castrated (N = 5 independent animals), 3. Castrated followed by Testosterone re-administration (Castrated-Testosterone independent animals) starting on day 189 (N = 4, N = 3 from day 203 to 252, N = 2 from day 252 to 273). Tumor scoring was performed weekly by routine palpation; values represent average calculation of the tumors of all animals per group (considering N = 2 tumors, left L and right R of each animal). Error bars represent SEM, calculated considering number of animals for each time point. Ordinary two-way ANOVA with Tukey’s multiple comparison correction was performed. (*) p = 0.0105 day 217, (**) p = 0.0025 day 224, (***) p = 0.0005 day 231, (****) p ≤ 0.0001 from day 238 onwards. e Top to bottom panels: Histological H&E staining of representative tumors from Castrated and Castrated-Testosterone hosts. Immunofluorescence staining for AR and CK5, CK8, NKX3.1 and CK8, CD44, and Ki67. DAPI marks the nuclei. Scale bars 50 μm. f Genomic analysis of PNPCa PDXs from intact and castrated animals, collecting samples at full regression (122 days) and after further testosterone replacement (84 days). g Principal component analysis of the gene expression of the 500 most variable genes. h Gene set enrichment analysis plot of statistically significant (adjusted p-value < 0.05) enrichment of HALLMARK pathways based on the differential expression analysis of the Castrated versus the Intact groups. NES, normalized enrichment score.
