Androgen Receptor: How splicing confers treatment resistance in prostate cancer

A splice variant of the androgen receptor that drives prostate cancer resistance translocates into the nucleus using a different mechanism from the full-length receptor and exhibits distinct molecular properties once inside.
  1. Prathyusha Konda
  2. Srinivas R Viswanathan  Is a corresponding author
  1. Dana-Farber Cancer Institute, Harvard Medical School, United States

Prostate cancer is the second leading cause of cancer-related deaths among men and claims over 350,000 lives worldwide each year. Most prostate cancers are driven by male hormones called androgens, which bind to and activate the androgen receptor (AR). Once an androgenic ligand, such as testosterone, binds to the androgen receptor, the receptor-ligand complex travels from the cytoplasm of the cell into its nucleus. Within the nucleus, the activated androgen receptor works as a transcription factor, which binds to DNA and triggers the cell to transcribe genes that are involved in prostate cancer progression. Therefore, androgen deprivation therapy, which lowers the levels of androgens in the body through drugs or surgical castration, represents an effective frontline treatment for prostate cancer.

While most prostate cancers initially shrink following treatment, resistance invariably emerges and the cancer relapses. This leads to the development of castration-resistant prostate cancer (CRPC), which can grow and spread to different parts of the body despite low levels of androgens. Multiple studies have shown that one of the most common molecular alterations seen in CRPC is the re-activation of signaling through the androgen receptor. This has led to the development of androgen receptor pathway inhibitors (ARPIs) that either block the part of the AR that binds to the ligand (thus preventing the receptor from becoming active) or interfere with the synthesis of androgens (Chen et al., 2004; Robinson et al., 2015; Visakorpi et al., 1995; Watson et al., 2015).

Unfortunately, many CRPCs develop resistance even to these drugs. It is thought that one of the mechanisms through which this resistance develops may be the production of splice variants of the androgen receptor. A splice variant is a version of a protein that results from ‘alternative splicing’ of the mRNA before it is translated into a protein. When an mRNA molecule is transcribed from the genome, it is spliced to remove sequences that do not code for the protein (known as introns), and to join together the remaining coding regions (known as exons). If this splicing process is modified, for example, due to a mutation, this can lead to different regions being spliced in or out of the mRNA. Consequently, different versions of the protein, or splice variants, are translated. In the case of the androgen receptor, splice variants that preserve the DNA-binding domain of the protein but lack the ligand-binding domain allow the protein to continue driving androgen receptor signaling even under low-androgen conditions (Figure 1; Dehm et al., 2008; Watson et al., 2015; Westaby et al., 2022).

Schematic of a full-length androgen receptor (AR) compared to splice variant AR-V7.

The full-length androgen receptor (AR-fl, left) binds to androgenic ligands via its ligand binding domain (LBD; dark grey rectangle) and activates the transcription of downstream genes. This form of the protein has a hinge domain (purple rectangle), which is required for the receptor to translocate into the nucleus, as well as a DNA binding domain (DBD; orange rectangle) and an N-terminal domain (NTD, green rectangle). Comparatively, a truncated splice variant of the receptor, called AR-V7 (right), only contains an NTD and a DBD followed by a cryptic exon (CE3; light grey rectangle), and lacks a hinge domain and an LBD. This version of the androgen receptor protein can activate downstream genes without binding to a ligand.

One important mechanism of resistance to ARPIs is the production of different androgen receptor splice variants. The most well-studied, AR-V7, is a truncated receptor that results when exons four to eight of the full-length mRNA sequence are missing. AR-V7 lacks the ligand binding domain of the full-length version, allowing it to signal in the absence of an androgenic ligand (Figure 1). The truncated receptor is both a biomarker of CRPC and a possible contributor to ARPI resistance (Antonarakis et al., 2014; Armstrong et al., 2019; Luo et al., 2018; Sharp et al., 2019; Westaby et al., 2022).

Interestingly, AR-V7 also lacks the hinge domain that the full-length receptor requires to translocate from the cytoplasm to the nucleus (Figure 1; Thadani-Mulero et al., 2014), leading to the question of how AR-V7 moves into the nucleus and exerts its transcriptional activity. Now, in eLife, Paraskevi Giannakakou and colleagues from Weill Cornell Medical College – including Seaho Kim, CheukMan Cherie Au, and Mohd Azrin Bin Jamalruddin as joint first authors – report how AR-V7 enters the nucleus (Kim et al., 2022).

Using microscopy to image live cells over time, Kim et al. demonstrated that, similar to the full-length receptor, AR-V7 is imported into the nucleus relatively quickly. However, AR-V7 does not rely on microtubules or importin – two protein complexes involved in the nuclear transport of the full-length version. Additionally, Kim et al. implicate the zinc finger (D-box) domain of AR-V7 in the nuclear import of the truncated receptor (which is not the case for the full-length receptor).

Fluorescence recovery after photobleaching, combined with other advanced microscopy techniques, demonstrated that AR-V7 is constantly moving within the nucleus, and does not stay in contact with the same region of DNA for long periods of time. This contrasts with the full-length counterpart (and other nuclear hormone receptors), which stay on the same region of DNA for prolonged durations, and as such, exhibit comparatively less movement within the nucleus.

These findings point to a ‘hit-and-run’ transcription model for AR-V7, in which it transiently binds to DNA sequences and recruits secondary transcription factors that keep the target gene active, even after it unbinds. While hit-and-run transcription has been typically associated with proteins that repress transcription, it has been proposed that AR-V7 may have repressive activity in CRPC (Cato et al., 2019). Intriguingly, Kim et al. found that AR-V7 also promotes the nuclear translocation of full-length androgen receptor without its ligand, although the exact mechanism remains unclear. It is also unknown how this may impact AR signaling. This information may be clinically relevant, as it is common for patients with CRPC to co-express the full-length receptor and spliced variant AR-V7 (Watson et al., 2010).

Taken together, these findings shed light on important distinctions between spliced AR-V7 and full-length androgen receptor, although several open questions remain. Further studies will be needed to identify which proteins transport AR-V7 into the nucleus. Additionally, it will be important to determine the link between the hit-and-run activity of AR-V7 and the function of this spliced receptor. Finally, the extent to which the mechanisms elucidated by Kim et al. apply to other spliced variants of the androgen receptor merits further investigation. More broadly, a thorough understanding of how the full-length androgen receptor is mechanistically distinct from its splice variants may provide opportunities to selectively block the variants from signaling in advanced prostate cancer.


Article and author information

Author details

  1. Prathyusha Konda

    Prathyusha Konda is at the Dana-Farber Cancer Institute, Harvard Medical School, Boston, United States

    Competing interests
    No competing interests declared
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6861-4442
  2. Srinivas R Viswanathan

    Srinivas R Viswanathan is at the Dana-Farber Cancer Institute, Harvard Medical School, Boston, United States

    For correspondence
    Competing interests
    No competing interests declared
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1856-3023

Publication history

  1. Version of Record published: August 23, 2022 (version 1)


© 2022, Konda and Viswanathan

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.


  • 489
    Page views
  • 122
  • 0

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Prathyusha Konda
  2. Srinivas R Viswanathan
Androgen Receptor: How splicing confers treatment resistance in prostate cancer
eLife 11:e82070.

Further reading

    1. Cancer Biology
    Judit Jimenez-Sainz, Joshua Mathew ... Ryan B Jensen
    Research Article Updated

    Pathogenic mutations in the BRCA2 tumor suppressor gene predispose to breast, ovarian, pancreatic, prostate, and other cancers. BRCA2 maintains genome stability through homology-directed repair (HDR) of DNA double-strand breaks (DSBs) and replication fork protection. Nonsense or frameshift mutations leading to truncation of the BRCA2 protein are typically considered pathogenic; however, missense mutations resulting in single amino acid substitutions can be challenging to functionally interpret. The majority of missense mutations in BRCA2 have been classified as Variants of Uncertain Significance (VUS) with unknown functional consequences. In this study, we identified three BRCA2 VUS located within the BRC repeat region to determine their impact on canonical HDR and fork protection functions. We provide evidence that S1221P and T1980I, which map to conserved residues in the BRC2 and BRC7 repeats, compromise the cellular response to chemotherapeutics and ionizing radiation, and display deficits in fork protection. We further demonstrate biochemically that S1221P and T1980I disrupt RAD51 binding and diminish the ability of BRCA2 to stabilize RAD51-ssDNA complexes. The third variant, T1346I, located within the spacer region between BRC2 and BRC3 repeats, is fully functional. We conclude that T1346I is a benign allele, whereas S1221P and T1980I are hypomorphic disrupting the ability of BRCA2 to fully engage and stabilize RAD51 nucleoprotein filaments. Our results underscore the importance of correctly classifying BRCA2 VUS as pathogenic variants can impact both future cancer risk and guide therapy selection during cancer treatment.

    1. Biochemistry and Chemical Biology
    2. Cancer Biology
    Stefania Monterisi, Johanna Michl ... Pawel Swietach
    Research Article Updated

    Growth of cancer cells in vitro can be attenuated by genetically inactivating selected metabolic pathways. However, loss-of-function mutations in metabolic pathways are not negatively selected in human cancers, indicating that these genes are not essential in vivo. We hypothesize that spontaneous mutations in ‘metabolic genes’ will not necessarily produce functional defects because mutation-bearing cells may be rescued by metabolite exchange with neighboring wild-type cells via gap junctions. Using fluorescent substances to probe intercellular diffusion, we show that colorectal cancer (CRC) cells are coupled by gap junctions assembled from connexins, particularly Cx26. Cells with genetically inactivated components of pH regulation (SLC9A1), glycolysis (ALDOA), or mitochondrial respiration (NDUFS1) could be rescued through access to functional proteins in co-cultured wild-type cells. The effect of diffusive coupling was also observed in co-culture xenografts. Rescue was largely dependent on solute exchange via Cx26 channels, a uniformly and constitutively expressed isoform in CRCs. Due to diffusive coupling, the emergent phenotype is less heterogenous than its genotype, and thus an individual cell should not be considered as the unit under selection, at least for metabolite-handling processes. Our findings can explain why certain loss-of-function mutations in genes ascribed as ‘essential’ do not influence the growth of human cancers.