Linking ligand-induced alterations in androgen receptor structure to differential gene expression: a first step in the rational design of selective androgen receptor modulators.
We have previously identified a family of novel androgen receptor (AR) ligands that, upon binding, enable AR to adopt structures distinct from that observed in the presence of canonical agonists. In this report, we describe the use of these compounds to establish a relationship between AR structure and biological activity with a view to defining a rational approach with which to identify useful selective AR modulators. To this end, we used combinatorial peptide phage display coupled with molecular dynamic structure analysis to identify the surfaces on AR that are exposed specifically in the presence of selected AR ligands. Subsequently, we used a DNA microarray analysis to demonstrate that differently conformed receptors facilitate distinct patterns of gene expression in LNCaP cells. Interestingly, we observed a complete overlap in the identity of genes expressed after treatment with mechanistically distinct AR ligands. However, it was differences in the kinetics of gene regulation that distinguished these compounds. Follow-up studies, in cell-based assays of AR action, confirmed the importance of these alterations in gene expression. Together, these studies demonstrate an important link between AR structure, gene expression, and biological outcome. This relationship provides a firm underpinning for mechanism-based screens aimed at identifying SARMs with useful clinical profiles.
Department of Pharmacology and Cancer Biology, Duke University Medical Center, Durham, North Carolina 27710, USA. donald.mcdonnell@duke.edu
Dmitri
D
Kazmin
Tatiana
T
Prytkova
C Edgar
CE
Cook
Russell
R
Wolfinger
Tzu-Ming
TM
Chu
David
D
Beratan
J D
JD
Norris
Ching-yi
CY
Chang
Donald P
DP
McDonnell
D000595:Amino Acid Sequence; D000728:Androgens; D001665:Binding Sites; D002460:Cell Line; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D066298:In Vitro Techniques; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D019151:Peptide Library; D040081:Protein Array Analysis; D011487:Protein Conformation; D011944:Receptors, Androgen / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000819:agonists / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013816:Thermodynamics
D000728:Androgens; D008024:Ligands; D019151:Peptide Library; D011944:Receptors, Androgen; D011994:Recombinant Proteins