Supplementary Material from Characterization of an Abiraterone Ultraresponsive Phenotype in Castration-Resistant Prostate Cancer Patient-Derived Xenografts
Supplementary Data Supplementary Table S1 Characteristics of the patient-derived xenograft models and protein expression by immunohistochemistry Supplementary Table S2. Primer sequences used in real-time PCR analysis Supplementary Table S3. Microarray gene expression analysis of steroidogenic genes in AA- versus vehicle-treated tumors upon AAresistance (end of study, EOS) Supplementary Table S4. Castration effects (vs. intact) on LuCaP PDX AR gene expression Supplementary Table S5. Upregulated genes in both ultraresponsive LuCaP 136CR and minimal responder LuCaP 35CR: lack of involvement in AR/GR signaling Supplementary Fig. S1. Treatment scheme and survival curves for AA on CRPC PDXs. Castration-resistant tumors were developed by either A, implanting castration-resistant tumors into castrated SCID mice (LuCaP 70CR), or B, implanting androgen-sensitive tumors into intact SCID mice and castrating mice when the tumor reached 100 mm3 (LuCaP 86.2CR). Castrated mice with castration-resistant tumors were treated orally with either vehicle or AA (0.5 mmol/kg/day) on a weekly schedule of 5 days on, 2 days off. Tumors were measured twice weekly. Mice were sacrificed and tumors were harvested on D7 or when tumors reached 1,000 mm3 (EOS). Supplementary Fig. S2. Differential gene expression associated with secreted proteins in the AA ultraresponder LuCaP 136CR. A, Chart showing 44% (68 genes) of 156 genes that were differentially expressed in the ultraresponder LuCaP 136CR (vs. intermediate responder LuCaP 96CR and minimal responder LuCaP 35CR) and were mapped into gene family coding for secreted proteins. Supervised clustering analyses showing 68 genes coding for secreted proteins at B, D7 and C, EOS. Red: high gene expression; green: low gene expression. Supplementary Fig. S3. Heat map showing the top 10 upregulated and top 10 downregulated genes representing secreted proteins in the LuCaP models at D7 and EOS. Yellow: high gene expression; blue: low gene expression. The heat map showed a homogeneous upregulation of the top 10 upregulated genes in LuCaP 136CR and a heterogeneous downregulation of the top 10 downregulated genes in LuCaP 136CR. Supplementary Fig. S4. qPCR confirmation on the top 10 downregulated genes of secreted proteins identified in the AA ultraresponder LuCaP 136CR versus intermediate responder LuCaP 77CR and LuCaP 96CR, and minimal responder LuCaP 35CR. Each data point represented one animal. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Supplementary Fig. S5. GSEA showing the association between differentially expressed genes upon AA treatment (D7) and resistance (EOS) in the LuCaP models with biological functions. False discovery rate <25% was considered statistically significant. Supplementary Fig. S6. Intratumoral androgen-related hormone level in LuCaP models upon AA resistance. Each data point represented one animal, n = 2-6.