Development of a High-Throughput Screening Assay for Small-Molecule Inhibitors of Androgen Receptor Splice Variants

TY - JOUR

T1 - Development of a High-Throughput Screening Assay for Small-Molecule Inhibitors of Androgen Receptor Splice Variants

AU - Monaghan, Amy E.

AU - Porter, Alison

AU - Hunter, Irene

AU - Morrison, Angus

AU - McElroy, Stuart P.

AU - McEwan, Iain J.

N1 - Funding Information: A.E.M. was supported by a BBSRC-CASE studentship together with support from IOmet Pharma (Nine Edinburgh Bioquarter, Edinburgh, Scotland, United Kingdom), NHS Grampian Endowments, and the charity Friends of Anchor. The high-throughput assay and screen were developed with support in kind from the Scottish Life Science Alliance (SULSA) assay development program. We are also grateful to support from Dr Alan Wise (IOmet Pharma) for advice and critical input. Publisher Copyright: © Amy E. Monaghan et al., 2022.

PY - 2022/4/18

Y1 - 2022/4/18

N2 - The role of the androgen receptor (AR) in the progression of prostate cancer (PCa) is well established and competitive inhibition of AR ligand binding domain (LBD) has been the mainstay of antiandrogen therapies for advanced and metastatic disease. However, the efficacy of such drugs is often limited by the emergence of resistance, mediated through point mutations and receptor splice variants lacking the AR-LBD. As a result, the prognosis for patients with malignant, castrate-resistant disease remains poor. The amino terminal domain (NTD) of the AR has been shown to be critical for AR function. Its modular activation function (AF-1) is important for both gene regulation and participation in protein-protein interactions. However, due to the intrinsically disordered structure of the domain, its potential as a candidate for therapeutic intervention has been generally overlooked. In this article, we describe the design and development of a functional cell-based assay aimed at identifying small-molecule inhibitors of the AR-NTD. We demonstrate the suitability of the assay for high-throughput screening platforms and validate two initial hits emerging from a small, targeted, library screen in PCa cells.

AB - The role of the androgen receptor (AR) in the progression of prostate cancer (PCa) is well established and competitive inhibition of AR ligand binding domain (LBD) has been the mainstay of antiandrogen therapies for advanced and metastatic disease. However, the efficacy of such drugs is often limited by the emergence of resistance, mediated through point mutations and receptor splice variants lacking the AR-LBD. As a result, the prognosis for patients with malignant, castrate-resistant disease remains poor. The amino terminal domain (NTD) of the AR has been shown to be critical for AR function. Its modular activation function (AF-1) is important for both gene regulation and participation in protein-protein interactions. However, due to the intrinsically disordered structure of the domain, its potential as a candidate for therapeutic intervention has been generally overlooked. In this article, we describe the design and development of a functional cell-based assay aimed at identifying small-molecule inhibitors of the AR-NTD. We demonstrate the suitability of the assay for high-throughput screening platforms and validate two initial hits emerging from a small, targeted, library screen in PCa cells.

KW - androgen receptor

KW - AR-vs

KW - castrate-resistant prostate cancer

KW - high-throughput screen

KW - intrinsically disordered structure

KW - prostate cancer

UR - https://www.scopus.com/pages/publications/85128801142

U2 - 10.1089/adt.2021.128

DO - 10.1089/adt.2021.128

M3 - Article

C2 - 35333596

AN - SCOPUS:85128801142

SN - 1540-658X

VL - 20

SP - 111

EP - 124

JO - ASSAY and Drug Development Technologies

JF - ASSAY and Drug Development Technologies

IS - 3

ER -