Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Muscarinic receptors promote castration-resistant growth of prostate cancer through a FAK–YAP signaling axis

Abstract

Prostate cancer (PCa) innervation contributes to the progression of PCa. However, the precise impact of innervation on PCa cells is still poorly understood. By focusing on muscarinic receptors, which are activated by the nerve-derived neurotransmitter acetylcholine, we show that muscarinic receptors 1 and 3 (m1 and m3) are highly expressed in PCa clinical specimens compared with all other cancer types, and that amplification or gain of their corresponding encoding genes (CHRM1 and CHRM3, respectively) represent a worse prognostic factor for PCa progression free survival. Moreover, m1 and m3 gene gain or amplification is frequent in castration-resistant PCa (CRPC) compared with hormone-sensitive PCa (HSPC) specimens. This was reflected in HSPC-derived cells, which show aberrantly high expression of m1 and m3 under androgen deprivation mimicking castration and androgen receptor inhibition. We also show that pharmacological activation of m1 and m3 signaling is sufficient to induce the castration-resistant growth of PCa cells. Mechanistically, we found that m1 and m3 stimulation induces YAP activation through FAK, whose encoding gene, PTK2 is frequently amplified in CRPC cases. Pharmacological inhibition of FAK and knockdown of YAP abolished m1 and m3-induced castration-resistant growth of PCa cells. Our findings provide novel therapeutic opportunities for muscarinic-signal-driven CRPC progression by targeting the FAK–YAP signaling axis.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Frequent overexpression of m1 and m3 in PCa and gene amplification and gain in CRPC.
Fig. 2: m1 and m3 are upregulated in CRPC cell lines, and activation of m1 and m3 induces castration-resistant growth of PCa cells.
Fig. 3: m1 and m3 signaling activate FAK and YAP, resulting in castration-resistant growth.
Fig. 4: Clinical impact of FAK gene expression on PCa.
Fig. 5: Pharmacological inhibition of FAK blocks carbachol-induced YAP activation and proliferation.

Similar content being viewed by others

References

  1. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2019. CA Cancer J Clin. 2019;69:7–34.

    Article  PubMed  Google Scholar 

  2. Crawford ED, Higano CS, Shore ND, Hussain M, Petrylak DP. Treating patients with metastatic castration resistant prostate cancer: a comprehensive review of available therapies. J Urol. 2015;194:1537–47.

    Article  PubMed  Google Scholar 

  3. Mauffrey P, Tchitchek N, Barroca V, Bemelmans A, Firlej V, Allory Y, et al. Progenitors from the central nervous system drive neurogenesis in cancer. Nature. 2019;569:672–8.

    Article  CAS  PubMed  Google Scholar 

  4. Zahalka AH, Arnal-Estape A, Maryanovich M, Nakahara F, Cruz CD, Finley LWS, et al. Adrenergic nerves activate an angio-metabolic switch in prostate cancer. Science. 2017;358:321–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Magnon C, Hall SJ, Lin J, Xue X, Gerber L, Freedland SJ, et al. Autonomic nerve development contributes to prostate cancer progression. Science. 2013;341:1236361.

    Article  PubMed  Google Scholar 

  6. Lee WY, Sun LM, Lin CL, Liang JA, Chang YJ, Sung FC, et al. Risk of prostate and bladder cancers in patients with spinal cord injury: a population-based cohort study. Urol Oncol. 2014;32:e51–7.

    Google Scholar 

  7. Patel N, Ngo K, Hastings J, Ketchum N, Sepahpanah F. Prevalence of prostate cancer in patients with chronic spinal cord injury. PM R. 2011;3:633–6.

    Article  PubMed  Google Scholar 

  8. Grytli HH, Fagerland MW, Fossa SD, Tasken KA. Association between use of beta-blockers and prostate cancer-specific survival: a cohort study of 3561 prostate cancer patients with high-risk or metastatic disease. Eur Urol. 2014;65:635–41.

    Article  CAS  PubMed  Google Scholar 

  9. Kaye JA, Margulis AV, Fortuny J, McQuay LJ, Plana E, Bartsch JL, et al. Cancer incidence after initiation of antimuscarinic medications for overactive bladder in the united kingdom: evidence for protopathic bias. Pharmacotherapy. 2017;37:673–83.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Wang N, Yao M, Xu J, Quan Y, Zhang K, Yang R, et al. Autocrine activation of CHRM3 promotes prostate cancer growth and castration resistance via CaM/CaMKK-mediated phosphorylation of Akt. Clin Cancer Res. 2015;21:4676–85.

    Article  CAS  PubMed  Google Scholar 

  11. Navone NM, Olive M, Ozen M, Davis R, Troncoso P, Tu SM, et al. Establishment of two human prostate cancer cell lines derived from a single bone metastasis. Clin Cancer Res. 1997;3:2493–2500.

    CAS  PubMed  Google Scholar 

  12. Chlenski A, Nakashiro K, Ketels KV, Korovaitseva GI, Oyasu R. Androgen receptor expression in androgen-independent prostate cancer cell lines. Prostate. 2001;47:66–75.

    Article  CAS  PubMed  Google Scholar 

  13. Gutkind JS, Novotny EA, Brann MR, Robbins KC. Muscarinic acetylcholine receptor subtypes as agonist-dependent oncogenes. Proc Natl Acad Sci USA. 1991;88:4703–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Tran C, Ouk S, Clegg NJ, Chen Y, Watson PA, Arora V, et al. Development of a second-generation antiandrogen for treatment of advanced prostate cancer. Science. 2009;324:787–90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Armbruster BN, Li X, Pausch MH, Herlitze S, Roth BL. Evolving the lock to fit the key to create a family of G protein-coupled receptors potently activated by an inert ligand. Proc Natl Acad Sci USA. 2007;104:5163–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Inoue A, Raimondi F, Kadji FMN, Singh G, Kishi T, Uwamizu A, et al. Illuminating G-protein-coupling selectivity of GPCRs. Cell. 2019;177:1933–47. e1925.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Feng X, Arang N, Rigiracciolo DC, Lee JS, Yeerna H, Wang Z, et al. A platform of synthetic lethal gene interaction networks reveals that the GNAQ uveal melanoma oncogene controls the Hippo pathway through FAK. Cancer Cell. 2019;35:457–72. e455.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. O’Hayre M, Vazquez-Prado J, Kufareva I, Stawiski EW, Handel TM, Seshagiri S, et al. The emerging mutational landscape of G proteins and G-protein-coupled receptors in cancer. Nat Rev Cancer. 2013;13:412–24.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  19. Wu V, Yeerna H, Nohata N, Chiou J, Harismendy O, Raimondi F, et al. Illuminating the Onco-GPCRome: novel G protein-coupled receptor-driven oncocrine networks and targets for cancer immunotherapy. J Biol Chem. 2019;294:11062–86.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Mendiratta P, Mostaghel E, Guinney J, Tewari AK, Porrello A, Barry WT, et al. Genomic strategy for targeting therapy in castration-resistant prostate cancer. J Clin Oncol. 2009;27:2022–9.

    Article  CAS  PubMed  Google Scholar 

  21. Drake JM, Graham NA, Lee JK, Stoyanova T, Faltermeier CM, Sud S. et al. Metastatic castration-resistant prostate cancer reveals intrapatient similarity and interpatient heterogeneity of therapeutic kinase targets. Proc Natl Acad Sci USA. 2013;110:E4762–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Arora VK, Schenkein E, Murali R, Subudhi SK, Wongvipat J, Balbas MD, et al. Glucocorticoid receptor confers resistance to antiandrogens by bypassing androgen receptor blockade. Cell. 2013;155:1309–22.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Schwartz S, Wongvipat J, Trigwell CB, Hancox U, Carver BS, Rodrik-Outmezguine V, et al. Feedback suppression of PI3Kalpha signaling in PTEN-mutated tumors is relieved by selective inhibition of PI3Kbeta. Cancer Cell. 2015;27:109–22.

    Article  CAS  PubMed  Google Scholar 

  24. Carver BS, Chapinski C, Wongvipat J, Hieronymus H, Chen Y, Chandarlapaty S, et al. Reciprocal feedback regulation of PI3K and androgen receptor signaling in PTEN-deficient prostate cancer. Cancer Cell. 2011;19:575–86.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Bluemn EG, Coleman IM, Lucas JM, Coleman RT, Hernandez-Lopez S, Tharakan R, et al. Androgen receptor pathway-independent prostate cancer is sustained through FGF signaling. Cancer Cell. 2017;32:474–89. e476.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Mannan Baig A, Khan NA, Effendi V, Rana Z, Ahmad HR, Abbas F. Differential receptor dependencies: expression and significance of muscarinic M1 receptors in the biology of prostate cancer. Anti-Cancer Drugs. 2017;28:75–87.

    Article  CAS  PubMed  Google Scholar 

  27. Luthin GR, Wang P, Zhou H, Dhanasekaran D, Ruggieri MR. Role of m1 receptor-G protein coupling in cell proliferation in the prostate. Life Sci. 1997;60:963–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Rayford W, Noble MJ, Austenfeld MA, Weigel J, Mebust WK, Shah GV. Muscarinic cholinergic receptors promote growth of human prostate cancer cells. Prostate. 1997;30:160–6.

    Article  CAS  PubMed  Google Scholar 

  29. Daaka Y. G proteins in cancer: the prostate cancer paradigm. Science’s STKE. 2004;2004:re2.

    PubMed  Google Scholar 

  30. Cao X, Qin J, Xie Y, Khan O, Dowd F, Scofield M, et al. Regulator of G-protein signaling 2 (RGS2) inhibits androgen-independent activation of androgen receptor in prostate cancer cells. Oncogene. 2006;25:3719–34.

    Article  CAS  PubMed  Google Scholar 

  31. Liu-Chittenden Y, Huang B, Shim JS, Chen Q, Lee SJ, Anders RA, et al. Genetic and pharmacological disruption of the TEAD-YAP complex suppresses the oncogenic activity of YAP. Genes Dev. 2012;26:1300–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Kuser-Abali G, Alptekin A, Lewis M, Garraway IP, Cinar B. YAP1 and AR interactions contribute to the switch from androgen-dependent to castration-resistant growth in prostate cancer. Nat Commun. 2015;6:8126.

    Article  PubMed  Google Scholar 

  33. Zhang L, Yang S, Chen X, Stauffer S, Yu F, Lele SM, et al. The hippo pathway effector YAP regulates motility, invasion, and castration-resistant growth of prostate cancer cells. Mol Cell Biol. 2015;35:1350–62.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Arnold JJ, Blinder KJ, Bressler NM, Bressler SB, Burdan A, Haynes L, et al. Acute severe visual acuity decrease after photodynamic therapy with verteporfin: case reports from randomized clinical trials-TAP and VIP report no. 3. Am J Ophthalmol. 2004;137:683–96.

    Article  PubMed  CAS  Google Scholar 

  35. Gibault F, Corvaisier M, Bailly F, Huet G, Melnyk P, Cotelle P. Non-photoinduced biological properties of verteporfin. Curr Med Chem. 2016;23:1171–84.

    Article  CAS  PubMed  Google Scholar 

  36. Zhang H, Ramakrishnan SK, Triner D, Centofanti B, Maitra D, Gyorffy B, et al. Tumor-selective proteotoxicity of verteporfin inhibits colon cancer progression independently of YAP1. Sci Signal. 2015;8:ra98.

    PubMed  PubMed Central  Google Scholar 

  37. Konstantinou EK, Notomi S, Kosmidou C, Brodowska K, Al-Moujahed A, Nicolaou F, et al. Verteporfin-induced formation of protein cross-linked oligomers and high molecular weight complexes is mediated by light and leads to cell toxicity. Sci Rep. 2017;7:46581.

    Article  PubMed  PubMed Central  Google Scholar 

  38. Roberts WG, Ung E, Whalen P, Cooper B, Hulford C, Autry C, et al. Antitumor activity and pharmacology of a selective focal adhesion kinase inhibitor, PF-562,271. Cancer Res. 2008;68:1935–44.

    Article  CAS  PubMed  Google Scholar 

  39. Slack-Davis JK, Hershey ED, Theodorescu D, Frierson HF, Parsons JT. Differential requirement for focal adhesion kinase signaling in cancer progression in the transgenic adenocarcinoma of mouse prostate model. Mol Cancer Ther. 2009;8:2470–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Schultze A, Fiedler W. Therapeutic potential and limitations of new FAK inhibitors in the treatment of cancer. Expert Opin Investig Drugs. 2010;19:777–88.

    Article  CAS  PubMed  Google Scholar 

  41. Sulzmaier FJ, Jean C, Schlaepfer DD. FAK in cancer: mechanistic findings and clinical applications. Nat Rev Cancer. 2014;14:598–610.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Bankhead P, Loughrey MB, Fernandez JA, Dombrowski Y, McArt DG, Dunne PD, et al. QuPath: Open source software for digital pathology image analysis. Sci Rep. 2017;7:16878.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  43. Gao J, Aksoy BA, Dogrusoz U, Dresdner G, Gross B, Sumer SO, et al. Integrative analysis of complex cancer genomics and clinical profiles using the cBioPortal. Sci Signal. 2013;6:pl1.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  44. Cerami E, Gao J, Dogrusoz U, Gross BE, Sumer SO, Aksoy BA, et al. The cBio cancer genomics portal: an open platform for exploring multidimensional cancer genomics data. Cancer Discov. 2012;2:401–4.

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

YG is supported by the JSPS Overseas Research Fellowships and the Uehara Memorial Foundation Research Fellowship. XF is supported by 111 Project of MOE (B14038) China, the National Natural Science Foundation (81402230, 81672677) China. MG is supported by FIRC-AIRC fellowship for abroad (Italian Foundation for cancer research). We thank La Jolla Institute Microscopy Core Facility for professional advice and guidance, in particular Zbigniew Mikulski. This work was supported by S10OD021831. YG initiated the study; YG and JSG designed the study and experiments; YG performed the genomic analyses; YG, TA, HI, NA, KA performed in vitro experiments, YG and MG performed immunofluorescence experiments, YG, NA and JSG prepared the manuscript, XF, ZW, NA and JSG provided advice and supervised the project. All authors discussed the results and reviewed the manuscript.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to J. Silvio Gutkind.

Ethics declarations

Conflict of interest

JSG is member of the Advisory Board of Oncoceutics and Domain Therapeutics.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Goto, Y., Ando, T., Izumi, H. et al. Muscarinic receptors promote castration-resistant growth of prostate cancer through a FAK–YAP signaling axis. Oncogene 39, 4014–4027 (2020). https://doi.org/10.1038/s41388-020-1272-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41388-020-1272-x

Search

Quick links