EG, was involved in idea development, experiment design, daily oral gavage drug treatments as well as tumor and tissue harvesting, RNA extraction and performing qPCR assays and data analysis. E.G. created Figure
3A and wrote first draft and edited manuscript.
MM prepared bicalutamide and vehicle control, performed caliper and total body weight measurements, was involved in tumor and tissue harvesting and processing and preparing RNA, protein extracts and tissue and tumor blocks for sectioning and IHC. MM performed qRT-PCR assays and analysis and PSA immunohistochemistry, collected digital Keyence microscope images, performed ImageScope analysis on Aperio Scanscope digitized images, applied final deconvolution algorithm, Conceived of data comparisons and created Figures
4 and
5. Contributed to writing and editing of manuscript.
AS, generated PCSD1 cells stably transduced with GFP-luciferase lentivirus and performed IVIS for Figure
1C. Prepared bicalutamide and vehicle control, Performed caliper and total body weight measurements, was involved in tumor and tissue harvesting, qPCR data analysis, writing and editing manuscript.
CNW was involved in experiment design, performed intra-femoral injections, monitored health and performed IVIS, contributed to editing of manuscript,
SCP was involved in in experiment design especially in terms of clinical relevance, oral gavage drug treatment, was involved in tumor and tissue harvesting, designed and performed correlational and statistical analyses of data, edited manuscript.
JRW, helped with experiment design and developed digital IHC analysis methodology, was involved in tumor and tissue harvesting, performed qPCR, edited manuscript.
WM, was involved in in experiment design especially in terms of clinical relevance, established oral gavage drug treatment protocol and performed drug treatments,
MAL was involved in in experiment design especially in terms of clinical relevance, was involved in developing digital IHC analysis methodology, and performed statistical analysis of digital IHC data,
TH, was involved in in experiment design especially in terms of clinical relevance, performed custom primer design, qPCR optimization and analysis,
OR, was involved in experiment design especially in terms of clinical relevance, performed custom primer design, qPCR optimization and analysis, edited manuscript and was involved in preparing Figure
2,
NAC contributed to manuscript writing, performed custom primer design, qPCR optimization and analysis, established protein extraction methods and performed Western blots, edited manuscript, A.A.K., is the orthopaedic surgeon specializing in pathologic fracture repair who provided patient bone metastasis, normal patient bone marrow and guided the experiment design and interpretation in terms of clinical insight and relevance to bone metastatic cancer,
CAMJ, is the PI of the study, wrote the manuscript, guided and coordinated all aspects of this translational research from obtaining patient specimens and discussing clinical needs for the patients with clinicians in order to developing models that most closely represent the disease in patients to directing the laboratory research for developing and testing the hypothesis. Formulated the hypothesis that prostate cancer is more resistant to androgen deprivation therapy in the bone microenvironment than in soft tissue. In addition, CAMJ performed SC injections, was involved in IF injections, performed oral gavage, performed patient sample preparation, xenograft tumor dissection and cell preparation for IF injections, RNA, DNA purification, tumor fixation. Planned, wrote and edited manuscript, designed figures, wrote and edited manuscript. All authors read and approved the final manuscript.