American Association for Cancer Research
Browse
10780432ccr162054-sup-169325_1_supp_3736881_cgc8q1.docx (2.69 MB)

Supplementary Material from Characterization of an Abiraterone Ultraresponsive Phenotype in Castration-Resistant Prostate Cancer Patient-Derived Xenografts

Download (2.69 MB)
journal contribution
posted on 2023-03-31, 19:46 authored by Hung-Ming Lam, Ryan McMullin, Holly M. Nguyen, Ilsa Coleman, Michael Gormley, Roman Gulati, Lisha G. Brown, Sarah K. Holt, Weimin Li, Deborah S. Ricci, Karin Verstraeten, Shibu Thomas, Elahe A. Mostaghel, Peter S. Nelson, Robert L. Vessella, Eva Corey

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.

Funding

Richard M. Lucas Foundation

Prostate Cancer Foundation

SU2C

NIH

PNW Prostate Cancer SPORE NIH

Department of Defense

Janssen Research & Development

History

ARTICLE ABSTRACT

Purpose: To identify the molecular signature associated with abiraterone acetate (AA) response and mechanisms underlying AA resistance in castration-resistant prostate cancer patient-derived xenografts (PDXs).Experimental Design: SCID mice bearing LuCaP 136CR, 77CR, 96CR, and 35CR PDXs were treated with AA. Tumor volume and prostate-specific antigen were monitored, and tumors were harvested 7 days after treatment or at end of study for gene expression and immunohistochemical studies.Results: Three phenotypic groups were observed based on AA response. An ultraresponsive phenotype was identified in LuCaP 136CR with significant inhibition of tumor progression and increased survival, intermediate responders LuCaP 77CR and LuCaP 96CR with a modest tumor inhibition and survival benefit, and LuCaP 35CR with minimal tumor inhibition and no survival benefit upon AA treatment. We identified a molecular signature of secreted proteins associated with the AA ultraresponsive phenotype. Upon resistance, AA ultraresponder LuCaP 136CR displayed reduced androgen receptor (AR) signaling and sustainably low nuclear glucocorticoid receptor (nGR) localization, accompanied by steroid metabolism alteration and epithelial–mesenchymal transition phenotype enrichment with increased expression of NF-κB–regulated genes; intermediate and minimal responders maintained sustained AR signaling and increased tumoral nGR localization.Conclusions: We identified a molecular signature of secreted proteins associated with AA ultraresponsiveness and sustained AR/GR signaling upon AA resistance in intermediate or minimal responders. These data will inform development of noninvasive biomarkers predicting AA response and suggest that further inhibition along the AR/GR signaling axis may be effective only in AA-resistant patients who are intermediate or minimal responders. These findings require verification in prospective clinical trials. Clin Cancer Res; 23(9); 2301–12. ©2016 AACR.

Usage metrics

    Clinical Cancer Research

    Licence

    Exports

    RefWorks
    BibTeX
    Ref. manager
    Endnote
    DataCite
    NLM
    DC