AR intragenic deletions linked to androgen receptor splice variant expression and activity in models of prostate cancer progression

Y. Li, T. H. Hwang, L. A. Oseth, A. Hauge, R. L. Vessella, S. C. Schmechel, B. Hirsch, K. B. Beckman, K. A. Silverstein, S. M. Dehm

Research output: Contribution to journalArticle

133 Citations (Scopus)

Abstract

Reactivation of the androgen receptor (AR) during androgen depletion therapy (ADT) underlies castration-resistant prostate cancer (CRPCa). Alternative splicing of the AR gene and synthesis of constitutively active COOH-terminally truncated AR variants lacking the AR ligand-binding domain has emerged as an important mechanism of ADT resistance in CRPCa. In a previous study, we demonstrated that altered AR splicing in CRPCa 22Rv1 cells was linked to a 35-kb intragenic tandem duplication of AR exon 3 and flanking sequences. In this study, we demonstrate that complex patterns of AR gene copy number imbalances occur in PCa cell lines, xenografts and clinical specimens. To investigate whether these copy number imbalances reflect AR gene rearrangements that could be linked to splicing disruptions, we carried out a detailed analysis of AR gene structure in the LuCaP 86.2 and CWR-R1 models of CRPCa. By deletion-spanning PCR, we discovered a 8579-bp deletion of AR exons 5, 6 and 7 in the LuCaP 86.2 xenograft, which provides a rational explanation for synthesis of the truncated AR v567es AR variant in this model. Similarly, targeted resequencing of the AR gene in CWR-R1 cells led to the discovery of a 48-kb deletion in AR intron 1. This intragenic deletion marked a specific CWR-R1 cell population with enhanced expression of the truncated AR-V7/AR3 variant, a high level of androgen-independent AR transcriptional activity and rapid androgen independent growth. Together, these data demonstrate that structural alterations in the AR gene are linked to stable gain-of-function splicing alterations in CRPCa.

Original languageEnglish (US)
Pages (from-to)4759-4767
Number of pages9
JournalOncogene
Volume31
Issue number45
DOIs
StatePublished - Nov 8 2012

Fingerprint

Androgen Receptors
Prostatic Neoplasms
Castration
Androgens
Heterografts
Genes
Exons
3' Flanking Region
Gene Dosage
Gene Rearrangement
Alternative Splicing
Introns

Keywords

  • Androgen receptor variants
  • AR alternative splicing
  • Castration-resistant
  • Intragenic rearrangement
  • Prostate cancer

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

AR intragenic deletions linked to androgen receptor splice variant expression and activity in models of prostate cancer progression. / Li, Y.; Hwang, T. H.; Oseth, L. A.; Hauge, A.; Vessella, R. L.; Schmechel, S. C.; Hirsch, B.; Beckman, K. B.; Silverstein, K. A.; Dehm, S. M.

In: Oncogene, Vol. 31, No. 45, 08.11.2012, p. 4759-4767.

Research output: Contribution to journalArticle

Li, Y, Hwang, TH, Oseth, LA, Hauge, A, Vessella, RL, Schmechel, SC, Hirsch, B, Beckman, KB, Silverstein, KA & Dehm, SM 2012, 'AR intragenic deletions linked to androgen receptor splice variant expression and activity in models of prostate cancer progression', Oncogene, vol. 31, no. 45, pp. 4759-4767. https://doi.org/10.1038/onc.2011.637
Li, Y. ; Hwang, T. H. ; Oseth, L. A. ; Hauge, A. ; Vessella, R. L. ; Schmechel, S. C. ; Hirsch, B. ; Beckman, K. B. ; Silverstein, K. A. ; Dehm, S. M. / AR intragenic deletions linked to androgen receptor splice variant expression and activity in models of prostate cancer progression. In: Oncogene. 2012 ; Vol. 31, No. 45. pp. 4759-4767.
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AU - Li, Y.

AU - Hwang, T. H.

AU - Oseth, L. A.

AU - Hauge, A.

AU - Vessella, R. L.

AU - Schmechel, S. C.

AU - Hirsch, B.

AU - Beckman, K. B.

AU - Silverstein, K. A.

AU - Dehm, S. M.

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N2 - Reactivation of the androgen receptor (AR) during androgen depletion therapy (ADT) underlies castration-resistant prostate cancer (CRPCa). Alternative splicing of the AR gene and synthesis of constitutively active COOH-terminally truncated AR variants lacking the AR ligand-binding domain has emerged as an important mechanism of ADT resistance in CRPCa. In a previous study, we demonstrated that altered AR splicing in CRPCa 22Rv1 cells was linked to a 35-kb intragenic tandem duplication of AR exon 3 and flanking sequences. In this study, we demonstrate that complex patterns of AR gene copy number imbalances occur in PCa cell lines, xenografts and clinical specimens. To investigate whether these copy number imbalances reflect AR gene rearrangements that could be linked to splicing disruptions, we carried out a detailed analysis of AR gene structure in the LuCaP 86.2 and CWR-R1 models of CRPCa. By deletion-spanning PCR, we discovered a 8579-bp deletion of AR exons 5, 6 and 7 in the LuCaP 86.2 xenograft, which provides a rational explanation for synthesis of the truncated AR v567es AR variant in this model. Similarly, targeted resequencing of the AR gene in CWR-R1 cells led to the discovery of a 48-kb deletion in AR intron 1. This intragenic deletion marked a specific CWR-R1 cell population with enhanced expression of the truncated AR-V7/AR3 variant, a high level of androgen-independent AR transcriptional activity and rapid androgen independent growth. Together, these data demonstrate that structural alterations in the AR gene are linked to stable gain-of-function splicing alterations in CRPCa.

AB - Reactivation of the androgen receptor (AR) during androgen depletion therapy (ADT) underlies castration-resistant prostate cancer (CRPCa). Alternative splicing of the AR gene and synthesis of constitutively active COOH-terminally truncated AR variants lacking the AR ligand-binding domain has emerged as an important mechanism of ADT resistance in CRPCa. In a previous study, we demonstrated that altered AR splicing in CRPCa 22Rv1 cells was linked to a 35-kb intragenic tandem duplication of AR exon 3 and flanking sequences. In this study, we demonstrate that complex patterns of AR gene copy number imbalances occur in PCa cell lines, xenografts and clinical specimens. To investigate whether these copy number imbalances reflect AR gene rearrangements that could be linked to splicing disruptions, we carried out a detailed analysis of AR gene structure in the LuCaP 86.2 and CWR-R1 models of CRPCa. By deletion-spanning PCR, we discovered a 8579-bp deletion of AR exons 5, 6 and 7 in the LuCaP 86.2 xenograft, which provides a rational explanation for synthesis of the truncated AR v567es AR variant in this model. Similarly, targeted resequencing of the AR gene in CWR-R1 cells led to the discovery of a 48-kb deletion in AR intron 1. This intragenic deletion marked a specific CWR-R1 cell population with enhanced expression of the truncated AR-V7/AR3 variant, a high level of androgen-independent AR transcriptional activity and rapid androgen independent growth. Together, these data demonstrate that structural alterations in the AR gene are linked to stable gain-of-function splicing alterations in CRPCa.

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