title: Phosphoproteomic profiling identifies focal adhesion kinase as a mediator of docetaxel resistance in castrate-resistant prostate cancer.
abstract: Docetaxel remains the standard-of-care for men diagnosed with metastatic castrate-resistant prostate cancer (CRPC). However, only approximately 50% of patients benefit from treatment and all develop docetaxel-resistant disease. Here, we characterize global perturbations in tyrosine kinase signaling associated with docetaxel resistance and thereby develop a potential therapeutic strategy to reverse this phenotype. Using quantitative mass spectrometry-based phosphoproteomics, we identified that metastatic docetaxel-resistant prostate cancer cell lines (DU145-Rx and PC3-Rx) exhibit increased phosphorylation of focal adhesion kinase (FAK) on Y397 and Y576, in comparison with parental controls (DU145 and PC3, respectively). Bioinformatic analyses identified perturbations in pathways regulating focal adhesions and the actin cytoskeleton and in protein-protein interaction networks related to these pathways in docetaxel-resistant cells. Treatment with the FAK tyrosine kinase inhibitor (TKI) PF-00562271 reduced FAK phosphorylation in the resistant cells, but did not affect cell viability or Akt phosphorylation. Docetaxel administration reduced FAK and Akt phosphorylation, whereas cotreatment with PF-00562271 and docetaxel resulted in an additive attenuation of FAK and Akt phosphorylation and overcame the chemoresistant phenotype. The enhanced efficacy of cotreatment was due to increased autophagic cell death, rather than apoptosis. These data strongly support that enhanced FAK activation mediates chemoresistance in CRPC, and identify a potential clinical niche for FAK TKIs, where coadministration with docetaxel may be used in patients with CRPC to overcome chemoresistance.
authors.affiliation: Corresponding Author: Roger J. Daly, Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Level 1, Building 77, Monash University, VIC 3800, Australia. Telephone: 61-3-990-29301; roger.daly@monash.edu.
authors.forename: Brian Y
authors.initials: BY
authors.lastname: Lee
authors.forename: Falko
authors.initials: F
authors.lastname: Hochgräfe
authors.forename: Hui-Ming
authors.initials: HM
authors.lastname: Lin
authors.forename: Lesley
authors.initials: L
authors.lastname: Castillo
authors.forename: Jianmin
authors.initials: J
authors.lastname: Wu
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Raftery
authors.forename: S
authors.initials: S
authors.lastname: Martin Shreeve
authors.forename: Lisa G
authors.initials: LG
authors.lastname: Horvath
authors.forename: Roger J
authors.initials: RJ
authors.lastname: Daly
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D000077143:Docetaxel; D019008:Drug Resistance, Neoplasm / Q000187:drug effects / Q000235:genetics; D051417:Focal Adhesion Kinase 1 / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008297:Male; D010750:Phosphoproteins / Q000302:isolation & purification / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D064129:Prostatic Neoplasms, Castration-Resistant / Q000188:drug therapy* / Q000235:genetics* / Q000473:pathology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D040901:Proteomics; D011743:Pyrimidines / Q000008:administration & dosage; D015398:Signal Transduction; D013449:Sulfonamides / Q000008:administration & dosage; D043823:Taxoids / Q000008:administration & dosage*
chemical_list: C573150:PF-00562271; D010750:Phosphoproteins; D011743:Pyrimidines; D013449:Sulfonamides; D043823:Taxoids; D000077143:Docetaxel; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1