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Neuropilin-2 regulates androgen-receptor transcriptional activity in advanced prostate cancer

Oncogene volume 41, pages 3747–3760 (2022)Cite this article

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Abstract

Aberrant transcriptional activity of androgen receptor (AR) is one of the dominant mechanisms for developing of castration-resistant prostate cancer (CRPC). Analyzing AR-transcriptional complex related to CRPC is therefore important towards understanding the mechanism of therapy resistance. While studying its mechanism, we observed that a transmembrane protein called neuropilin-2 (NRP2) plays a contributory role in forming a novel AR-transcriptional complex containing nuclear pore proteins. Using immunogold electron microscopy, high-resolution confocal microscopy, chromatin immunoprecipitation, proteomics, and other biochemical techniques, we delineated the molecular mechanism of how a specific splice variant of NRP2 becomes sumoylated upon ligand stimulation and translocates to the inner nuclear membrane. This splice variant of NRP2 then stabilizes the complex between AR and nuclear pore proteins to promote CRPC specific gene expression. Both full-length and splice variants of AR have been identified in this specific transcriptional complex. In vitro cell line-based assays indicated that depletion of NRP2 not only destabilizes the AR-nuclear pore protein interaction but also inhibits the transcriptional activities of AR. Using an in vivo bone metastasis model, we showed that the inhibition of NRP2 led to the sensitization of CRPC cells toward established anti-AR therapies such as enzalutamide. Overall, our finding emphasize the importance of combinatorial inhibition of NRP2 and AR as an effective therapeutic strategy against treatment refractory prostate cancer.

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Fig. 1: NRP2 localization detected within the nucleus.
Fig. 2: NRP2 present in the nuclear membrane of the cell.
Fig. 3: VEGF-C-mediated retrograde transport and post-translational SUMOylation of NRP2B.
Fig. 4: NRP2 Mass-Spectrometry predicted its interactions with various nuclear-associated proteins.
Fig. 5: Analysis of AR-NRP2 regulated gene expression.
Fig. 6: AR-DNA interaction decreases following NRP2 depletion.
Fig. 7: Inhibition of nuclear transport of NRP2 increased AR-targeted therapeutic efficacy.

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Acknowledgements

The authors thank all stuff members of the Advanced Microscopy Core and Genomics Core Facility of UNMC. We also extend our sincere thanks to the UNMC Bioinformatics and Bio-statistician Core for their support to analyze the data. The raw data has been deposited in Gene Expression Omnibus with the accession number GSE205150.

Funding

This work was supported by grants for SD (1R21CA241234-01, NE-LB506, Lageschulte Fund), KD (R01CA182435, R01CA239343, DoD W81XWH2110628), MHM and LHH (DFG project number 273676790), and MHM (DFG project number 416001651). MM is funded by the Rudolf-Becker-Foundation for his professorship. The construction of the prostate cancer tissue microarray was funded by the DFG Forschergruppe-1586 SKELMET to LCH and SF.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha, NE, USA

    Samikshan Dutta, Navatha Shree Polavaram, Ridwan Islam, Sreyashi Bhattacharya, Sanika Bodas, Sohini Roy, David L. Klinkebiel, Surinder K. Batra & Kaustubh Datta

  2. Rudolf Becker Laboratory for Prostate Cancer Research, Medical Faculty, University of Bonn, Bonn, Germany

    Thomas Mayr & Michael H. Muders

  3. Institute of Pathology, Medical Faculty, University of Bonn, Bonn, Germany

    Thomas Mayr, Marieta I. Toma & Michael H. Muders

  4. Institute of Pathology, Technische Universitaet Dresden, Dresden, Germany

    Thomas Mayr, Marieta I. Toma, Gustavo B. Baretton & Michael H. Muders

  5. Department of Agronomy and Horticulture, University of Nebraska-Lincoln, Lincoln, NE, USA

    Sophie Alvarez Y. Albala

  6. Department of Cell Biology, UT Southwestern Medical Center, Dallas, TX, USA

    Anza Darehshouri

  7. Department of Urology, Technische Universitaet Dresden, Dresden, Germany

    Angelika Borkowetz, Susanne Fuessel & Manfred Wirth

  8. Division of Endocrinology and Metabolic Bone Diseases, Department of Medicine III, Technische Universitaet Dresden, Dresden, Germany

    Stefanie Conrad & Lorenz C. Hofbauer

  9. Center for Healthy Aging, Technische Universitaet Dresden, Dresden, Germany

    Stefanie Conrad & Lorenz C. Hofbauer

  10. German Cancer Consortium (DKTK), Partner Site Dresden, German Research Center (DKFZ), Heidelberg, Germany

    Gustavo B. Baretton & Lorenz C. Hofbauer

  11. Tumor and Normal Tissue Bank of the University Cancer Center (UCC), University Hospital and Faculty of Medicine, Technische Universitaet Dresden, Dresden, Germany

    Gustavo B. Baretton

  12. Department of Biochemistry and Molecular Medicine, University of California Davis, Davis, CA, USA

    Paramita Ghosh

  13. The Brady Urological Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Kenneth J. Pienta

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  1. Samikshan Dutta
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Contributions

SD and KD has designed the project and SD performed most of the work. NSP, RI, SB, SB, SR have assisted some of the work. TM has developed the NRP2-HA-tagged plasmid. SAA did the mass spec. AD took the electron microscopic images. MI, AB, SC, SF, MW, GBB, LCH, MHM were involved in TMA development and staining of NRP2. PG help us in promoter assay. PG, KJP, SKB and MHM critically evaluated the work and time to time provide there suggestion. DLK performed and analyzed the ChIP-seq.

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Correspondence to Samikshan Dutta or Kaustubh Datta.

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Dutta, S., Polavaram, N.S., Islam, R. et al. Neuropilin-2 regulates androgen-receptor transcriptional activity in advanced prostate cancer. Oncogene 41, 3747–3760 (2022). https://doi.org/10.1038/s41388-022-02382-y

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