title: Functional analysis of androgen receptor mutations that confer anti-androgen resistance identified in circulating cell-free DNA from prostate cancer patients.
abstract: BACKGROUND
  The androgen receptor (AR) is a pivotal drug target for the treatment of prostate cancer, including its lethal castration-resistant (CRPC) form. All current non-steroidal AR antagonists, such as hydroxyflutamide, bicalutamide, and enzalutamide, target the androgen binding site of the receptor, competing with endogenous androgenic steroids. Several AR mutations in this binding site have been associated with poor prognosis and resistance to conventional prostate cancer drugs. In order to develop an effective CRPC therapy, it is crucial to understand the effects of these mutations on the functionality of the AR and its ability to interact with endogenous steroids and conventional AR inhibitors.
  
  
  RESULTS
  We previously utilized circulating cell-free DNA (cfDNA) sequencing technology to examine the AR gene for the presence of mutations in CRPC patients. By modifying our sequencing and data analysis approaches, we identify four additional single AR mutations and five mutation combinations associated with CRPC. Importantly, we conduct experimental functionalization of all the AR mutations identified by the current and previous cfDNA sequencing to reveal novel gain-of-function scenarios. Finally, we evaluate the effect of a novel class of AR inhibitors targeting the binding function 3 (BF3) site on the activity of CRPC-associated AR mutants.
  
  
  CONCLUSIONS
  This work demonstrates the feasibility of a prognostic and/or diagnostic platform combining the direct identification of AR mutants from patients' serum, and the functional characterization of these mutants in order to provide personalized recommendations regarding the best future therapy.
authors.affiliation: Vancouver Prostate Centre, University of British Columbia, 2660 Oak St., Vancouver, BC, V6H 3Z6, Canada. nlallous@prostatecentre.com.
authors.forename: Nada
authors.initials: N
authors.lastname: Lallous
authors.affiliation: Vancouver Prostate Centre, University of British Columbia, 2660 Oak St., Vancouver, BC, V6H 3Z6, Canada. svolik@prostatecentre.com.|Laboratory for Advanced Genome Analysis (LAGA), Vancouver Prostate Centre, 2660 Oak St., Vancouver, BC, V6H 3Z6, Canada. svolik@prostatecentre.com.
authors.forename: Stanislav V
authors.initials: SV
authors.lastname: Volik
authors.affiliation: Vancouver Prostate Centre, University of British Columbia, 2660 Oak St., Vancouver, BC, V6H 3Z6, Canada. sawrey@prostatecentre.com.
authors.forename: Shannon
authors.initials: S
authors.lastname: Awrey
authors.affiliation: Vancouver Prostate Centre, University of British Columbia, 2660 Oak St., Vancouver, BC, V6H 3Z6, Canada. eleblanc@prostatecentre.com.
authors.forename: Eric
authors.initials: E
authors.lastname: Leblanc
authors.affiliation: Vancouver Prostate Centre, University of British Columbia, 2660 Oak St., Vancouver, BC, V6H 3Z6, Canada. rst2@sfu.ca.
authors.forename: Ronnie
authors.initials: R
authors.lastname: Tse
authors.affiliation: Vancouver Prostate Centre, University of British Columbia, 2660 Oak St., Vancouver, BC, V6H 3Z6, Canada. jmurillo@prostatecentre.com.
authors.forename: Josef
authors.initials: J
authors.lastname: Murillo
authors.affiliation: Vancouver Prostate Centre, University of British Columbia, 2660 Oak St., Vancouver, BC, V6H 3Z6, Canada. ksingh@prostatecentre.com.
authors.forename: Kriti
authors.initials: K
authors.lastname: Singh
authors.affiliation: Department of Medical Oncology, BC Cancer Agency, 600 West 10th Avenue, Vancouver, BC, V5Z 4E6, Canada. Arun.Azad@bccancer.bc.ca.
authors.forename: Arun A
authors.initials: AA
authors.lastname: Azad
authors.affiliation: Vancouver Prostate Centre, University of British Columbia, 2660 Oak St., Vancouver, BC, V6H 3Z6, Canada. awyatt@prostatecentre.com.
authors.forename: Alexander W
authors.initials: AW
authors.lastname: Wyatt
authors.affiliation: Vancouver Prostate Centre, University of British Columbia, 2660 Oak St., Vancouver, BC, V6H 3Z6, Canada. slebihan@prostatecentre.com.|Laboratory for Advanced Genome Analysis (LAGA), Vancouver Prostate Centre, 2660 Oak St., Vancouver, BC, V6H 3Z6, Canada. slebihan@prostatecentre.com.
authors.forename: Stephane
authors.initials: S
authors.lastname: LeBihan
authors.affiliation: Department of Medical Oncology, BC Cancer Agency, 600 West 10th Avenue, Vancouver, BC, V5Z 4E6, Canada. kchi@bccancer.bc.ca.
authors.forename: Kim N
authors.initials: KN
authors.lastname: Chi
authors.affiliation: Vancouver Prostate Centre, University of British Columbia, 2660 Oak St., Vancouver, BC, V6H 3Z6, Canada. mgleave@prostatecentre.com.
authors.forename: Martin E
authors.initials: ME
authors.lastname: Gleave
authors.affiliation: Vancouver Prostate Centre, University of British Columbia, 2660 Oak St., Vancouver, BC, V6H 3Z6, Canada. prennie@prostatecentre.com.
authors.forename: Paul S
authors.initials: PS
authors.lastname: Rennie
authors.affiliation: Vancouver Prostate Centre, University of British Columbia, 2660 Oak St., Vancouver, BC, V6H 3Z6, Canada. ccollins@prostatecentre.com.|Laboratory for Advanced Genome Analysis (LAGA), Vancouver Prostate Centre, 2660 Oak St., Vancouver, BC, V6H 3Z6, Canada. ccollins@prostatecentre.com.
authors.forename: Colin C
authors.initials: CC
authors.lastname: Collins
authors.affiliation: Vancouver Prostate Centre, University of British Columbia, 2660 Oak St., Vancouver, BC, V6H 3Z6, Canada. artc@interchange.ubc.ca.
authors.forename: Artem
authors.initials: A
authors.lastname: Cherkasov
mesh_terms: D059002:Androgen Receptor Antagonists / Q000627:therapeutic use; D000813:Anilides / Q000494:pharmacology; D001549:Benzamides; D004247:DNA / Q000097:blood / Q000235:genetics*; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D005485:Flutamide / Q000031:analogs & derivatives / Q000494:pharmacology; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008297:Male; D009154:Mutation / Q000235:genetics; D009570:Nitriles / Q000494:pharmacology; D010669:Phenylthiohydantoin / Q000031:analogs & derivatives / Q000494:pharmacology; D064129:Prostatic Neoplasms, Castration-Resistant / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D011944:Receptors, Androgen / Q000235:genetics*; D014105:Tosyl Compounds / Q000494:pharmacology
chemical_list: C410798:AR protein, human; D059002:Androgen Receptor Antagonists; D000813:Anilides; D001549:Benzamides; D009570:Nitriles; D011944:Receptors, Androgen; D014105:Tosyl Compounds; D010669:Phenylthiohydantoin; C014290:hydroxyflutamide; D005485:Flutamide; D004247:DNA; C540278:enzalutamide; C053541:bicalutamide
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