Genetic manipulation of autonomic nerve fiber innervation and activity and its effect on breast cancer progression


The effects of autonomic innervation of tumors on tumor growth remain unclear. Here we developed a series of genetic techniques to manipulate autonomic innervation in a tumor- and fiber-type-specific manner in mice with human breast cancer xenografts and in rats with chemically induced breast tumors. Breast cancer growth and progression were accelerated following stimulation of sympathetic nerves in tumors, but were reduced following stimulation of parasympathetic nerves. Tumor-specific sympathetic denervation suppressed tumor growth and downregulated the expression of immune checkpoint molecules (programed death-1 (PD-1), programed death ligand-1 (PD-L1), and FOXP3) to a greater extent than with pharmacological α- or β-adrenergic receptor blockers. Genetically induced simulation of parasympathetic innervation of tumors decreased PD-1 and PD-L1 expression. In humans, a retrospective analysis of breast cancer specimens from 29 patients revealed that increased sympathetic and decreased parasympathetic nerve density in tumors were associated with poor clinical outcomes and correlated with higher expression of immune checkpoint molecules. These findings suggest that autonomic innervation of tumors regulates breast cancer progression.

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Fig. 1: Sympathetic innervation of tumors accelerates the growth and progression of human breast cancer cell xenografts in mice.
Fig. 2: Sympathetic nerve denervation of tumors, rather than injections of adrenergic receptor blockers, suppresses the growth and progression of human breast cancer cell xenografts.
Fig. 3: Sympathetic innervation of tumors accelerates the growth of chemically induced breast cancer via the release of neurotransmitters.
Fig. 4: Sympathetic nerve denervation, rather than injections of adrenergic receptor blockers, suppresses immune checkpoint molecule expression in the tumor microenvironment.
Fig. 5: Parasympathetic innervation of tumors decelerates the growth and progression of human breast cancer cell xenografts in mice.
Fig. 6: Parasympathetic innervation of tumors decelerates the growth of chemically induced breast cancer and suppresses immune checkpoint molecule expression.
Fig. 7: Abundance of sympathetic and sparcity of parasympathetic nerve fibers in individuals with breast cancer with recurrence.
Fig. 8: Expression of immune checkpoint molecules positively and negatively correlates with sympathetic and parasympathetic nerve density, respectively, in human breast cancer samples.

Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Code availability

No custom code was generated for this study.


  1. 1.

    Cole, S. W., Nagaraja, A. S., Lutgendorf, S. K., Green, P. A. & Sood, A. K. Sympathetic nervous system regulation of the tumour microenvironment. Nat. Rev. Cancer 15, 563–572 (2015).

    CAS  Article  Google Scholar 

  2. 2.

    Hanoun, M., Maryanovich, M., Arnal-Estape, A. & Frenette, P. S. Neural regulation of hematopoiesis, inflammation, and cancer. Neuron 86, 360–373 (2015).

    CAS  Article  Google Scholar 

  3. 3.

    Chida, Y., Hamer, M., Wardle, J. & Steptoe, A. Do stress-related psychosocial factors contribute to cancer incidence and survival? Nat. Clin. Pract. Oncol. 5, 466–475 (2008).

    Article  Google Scholar 

  4. 4.

    Thaker, P. H. et al. Chronic stress promotes tumor growth and angiogenesis in a mouse model of ovarian carcinoma. Nat. Med 12, 939–944 (2006).

    CAS  Article  Google Scholar 

  5. 5.

    Schuller, H. M., Al-Wadei, H. A., Ullah, M. F. & Plummer, H. K. III. Regulation of pancreatic cancer by neuropsychological stress responses: a novel target for intervention. Carcinogenesis 33, 191–196 (2011).

    Article  Google Scholar 

  6. 6.

    Sloan, E. K. et al. The sympathetic nervous system induces a metastatic switch in primary breast cancer. Cancer Res. 70, 7042–7052 (2010).

    CAS  Article  Google Scholar 

  7. 7.

    Barron, T. I., Connolly, R. M., Sharp, L., Bennett, K. & Visvanathan, K. Beta blockers and breast cancer mortality: a population- based study. J. Clin. Oncol. 29, 2635–2644 (2011).

    CAS  Article  Google Scholar 

  8. 8.

    Cardwell, C. R., Coleman, H. G., Murray, L. J., Entschladen, F. & Powe, D. G. Beta-blocker usage and breast cancer survival: a nested case-control study within a UK clinical practice research datalink cohort. Int. J. Epidemiol. 42, 1852–1861 (2014).

    Article  Google Scholar 

  9. 9.

    Melhem-Bertrandt, A. et al. Beta-blocker use is associated with improved relapse-free survival in patients with triple-negative breast cancer. J. Clin. Oncol. 29, 2645–2652 (2011).

    CAS  Article  Google Scholar 

  10. 10.

    Shaashua, L. et al. Perioperative COX-2 and beta-adrenergic blockade improves metastatic biomarkers in breast cancer patients in a phase-II randomized trial. Clin. Cancer Res. 23, 4651–4661 (2017).

    CAS  Article  Google Scholar 

  11. 11.

    Grytli, H. H., Fagerland, M. W., Fossa, S. D. & Tasken, K. A. 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. 65, 635–641 (2013).

    Article  Google Scholar 

  12. 12.

    Sorensen, G. V. et al. Use of beta-blockers, angiotensin-converting enzyme inhibitors, angiotensin II receptor blockers, and risk of breast cancer recurrence: a Danish nationwide prospective cohort study. J. Clin. Oncol. 31, 2265–2272 (2013).

    Article  Google Scholar 

  13. 13.

    Magnon, C. et al. Autonomic nerve development contributes to prostate cancer progression. Science 341, 1236361 (2013).

    Article  Google Scholar 

  14. 14.

    Bellinger, D. L. & Lorton, D. Autonomic regulation of cellular immune function. Auton. Neurosci. 182, 15–41 (2014).

    CAS  Article  Google Scholar 

  15. 15.

    Chen, L. & Han, X. Anti-PD-1/PD-L1 therapy of human cancer: past, present, and future. J. Clin. Invest. 125, 3384–3391 (2015).

    Article  Google Scholar 

  16. 16.

    Tanaka, A. & Sakaguchi, S. Regulatory T cells in cancer immunotherapy. Cell Res. 27, 109–118 (2016).

    Article  Google Scholar 

  17. 17.

    Felten, D. L., Hall, P. V., Campbell, R. L. & Kalsbeck, J. E. A histochemical investigation of catecholamines in spinal cord injury. J. Neural. Transm. 39, 209–221 (1976).

    CAS  Article  Google Scholar 

  18. 18.

    Hollis, E. R. 2nd, Kadoya, K., Hirsch, M., Samulski, R. J. & Tuszynski, M. H. Efficient retrograde neuronal transduction utilizing self-complementary AAV1. Mol. Ther. 16, 296–301 (2008).

    CAS  Article  Google Scholar 

  19. 19.

    Pirozzi, M. et al. Intramuscular viral delivery of paraplegin rescues peripheral axonopathy in a model of hereditary spastic paraplegia. J. Clin. Invest. 116, 202–208 (2006).

    CAS  Article  Google Scholar 

  20. 20.

    Irie, K. et al. Comparative study of the gating motif and C-type inactivation in prokaryotic voltage-gated sodium channels. J. Biol. Chem. 285, 3685–3694 (2009).

    Article  Google Scholar 

  21. 21.

    Ren, D. et al. A prokaryotic voltage-gated sodium channel. Science 294, 2372–2375 (2001).

    CAS  Article  Google Scholar 

  22. 22.

    Lin, C. W. et al. Genetically increased cell-intrinsic excitability enhances neuronal integration into adult brain circuits. Neuron 65, 32–39 (2010).

    CAS  Article  Google Scholar 

  23. 23.

    Sakamoto, M. et al. Continuous neurogenesis in the adult forebrain is required for innate olfactory responses. Proc. Natl Acad. Sci. USA 108, 8479–8484 (2011).

    CAS  Article  Google Scholar 

  24. 24.

    Ubil, E. et al. Mesenchymal–endothelial transition contributes to cardiac neovascularization. Nature 514, 585–590 (2014).

    CAS  Article  Google Scholar 

  25. 25.

    Kinoshita, M. et al. Genetic dissection of the circuit for hand dexterity in primates. Nature 487, 235–238 (2012).

    CAS  Article  Google Scholar 

  26. 26.

    Lamkin, D. M. et al. Alpha2-adrenergic blockade mimics the enhancing effect of chronic stress on breast cancer progression. Psychoneuroendocrinology 51, 262–270 (2014).

    Article  Google Scholar 

  27. 27.

    Kamiya, A., Kawada, T., Shimizu, S. & Sugimachi, M. Closed-loop spontaneous baroreflex transfer function is inappropriate for system identification of neural arc but partly accurate for peripheral arc: predictability analysis. J. Physiol. 589, 1769–1790 (2011).

    CAS  PubMed  PubMed Central  Google Scholar 

  28. 28.

    Struyker-Boudier, H. A., van Essen, H., Nievelstein, H. M. & Smits, J. F. Role of baroreflex activation in the regional hemodynamic effects of the beta-blockers tertatolol and propranolol in conscious spontaneously hypertensive rats. Am. J. Nephrol. 6 (Suppl. 2), 25–29 (1986).

    Article  Google Scholar 

  29. 29.

    Kennedy, J. D., Pierce, C. W. & Lake, J. P. Extrathymic T cell maturation. Phenotypic analysis of T cell subsets in nude mice as a function of age. J. Immunol. 148, 1620–1629 (1992).

    CAS  PubMed  Google Scholar 

  30. 30.

    Nakajima, C. et al. A role of interferon-gamma (IFN-gamma) in tumor immunity: T cells with the capacity to reject tumor cells are generated but fail to migrate to tumor sites in IFN-gamma-deficient mice. Cancer Res. 61, 3399–3405 (2001).

    CAS  PubMed  Google Scholar 

  31. 31.

    Zanetti, M. Tapping CD4 T cells for cancer immunotherapy: the choice of personalized genomics. J. Immunol. 194, 2049–2056 (2015).

    CAS  Article  Google Scholar 

  32. 32.

    Bhat, P., Leggatt, G., Waterhouse, N. & Frazer, I. H. Interferon-gamma derived from cytotoxic lymphocytes directly enhances their motility and cytotoxicity. Cell Death Dis. 8, e2836 (2017).

    CAS  Article  Google Scholar 

  33. 33.

    Ligocki, A. J., Brown, J. R. & Niederkorn, J. Y. Role of interferon-gamma and cytotoxic T lymphocytes in intraocular tumor rejection. J. Leukoc. Biol. 99, 735–747 (2016).

    CAS  Article  Google Scholar 

  34. 34.

    Zhou, J., Ma, P., Li, J., Cui, X. & Song, W. Improvement of the cytotoxic T lymphocyte response against hepatocellular carcinoma by transduction of cancer cells with an adeno-associated virus carrying the interferon-gamma gene. Mol. Med. Rep. 13, 3197–3205 (2016).

    CAS  Article  Google Scholar 

  35. 35.

    Asamoto, M. et al. Transgenic rats carrying human c-Ha-ras proto-oncogenes are highly susceptible to N-methyl-N-nitrosourea mammary carcinogenesis. Carcinogenesis 21, 243–249 (2000).

    CAS  Article  Google Scholar 

  36. 36.

    Zhou, X., Vink, M., Klaver, B., Berkhout, B. & Das, A. T. Optimization of the Tet-On system for regulated gene expression through viral evolution. Gene Ther. 13, 1382–1390 (2006).

    CAS  Article  Google Scholar 

  37. 37.

    Gossen, M. & Bujard, H. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc. Natl Acad. Sci. USA 89, 5547–5551 (1992).

    CAS  Article  Google Scholar 

  38. 38.

    Guy, C. T., Cardiff, R. D. & Muller, W. J. Induction of mammary tumors by expression of polyomavirus middle T oncogene: a transgenic mouse model for metastatic disease. Mol. Cell Biol. 12, 954–961 (1992).

    CAS  Article  Google Scholar 

  39. 39.

    Bucsek, M. J. et al. Beta-adrenergic signaling in mice housed at standard temperatures suppresses an effector phenotype in CD8+ T cells and undermines checkpoint inhibitor therapy. Cancer Res. 77, 5639–5651 (2017).

    CAS  Article  Google Scholar 

  40. 40.

    Perron, L., Bairati, I., Harel, F. & Meyer, F. Antihypertensive drug use and the risk of prostate cancer (Canada). Cancer Causes Control 15, 535–541 (2004).

    Article  Google Scholar 

  41. 41.

    Powe, D. G. et al. Beta-blocker drug therapy reduces secondary cancer formation in breast cancer and improves cancer specific survival. Oncotarget 1, 628–638 (2010).

    Article  Google Scholar 

  42. 42.

    Hicks, B. M., Murray, L. J., Powe, D. G., Hughes, C. M. & Cardwell, C. R. Beta-blocker usage and colorectal cancer mortality: a nested case-control study in the UK Clinical Practice Research Datalink cohort. Ann. Oncol. 24, 3100–3106 (2013).

    CAS  Article  Google Scholar 

  43. 43.

    McCourt, C. et al. Beta-blocker usage after malignant melanoma diagnosis and survival: a population-based nested case-control study. Br. J. Dermatol. 170, 930–938 (2014).

    CAS  Article  Google Scholar 

  44. 44.

    Hayakawa, Y. et al. Nerve growth factor promotes gastric tumorigenesis through aberrant cholinergic signaling. Cancer Cell 31, 21–34 (2017).

    CAS  Article  Google Scholar 

  45. 45.

    Zhao, C. M. et al. Denervation suppresses gastric tumorigenesis. Sci. Transl Med. 6, 250ra115 (2014).

    Article  Google Scholar 

  46. 46.

    Zahalka, A. H. et al. Adrenergic nerves activate an angio-metabolic switch in prostate cancer. Science 358, 321–326 (2017).

    CAS  Article  Google Scholar 

  47. 47.

    Oh, M. S., Hong, S. J., Huh, Y. & Kim, K. S. Expression of transgenes in midbrain dopamine neurons using the tyrosine hydroxylase promoter. Gene Ther. 16, 437–440 (2009).

    CAS  Article  Google Scholar 

  48. 48.

    Eto, K. et al. Enhanced GABAergic activity in the mouse primary somatosensory cortex is insufficient to alleviate chronic pain behavior with reduced expression of neuronal potassium-chloride cotransporter. J. Neurosci. 32, 16552–16559 (2012).

    CAS  Article  Google Scholar 

  49. 49.

    Borg, M. L. et al. Hypothalamic neurogenesis is not required for the improved insulin sensitivity following exercise training. Diabetes 63, 3647–3658 (2014).

    CAS  Article  Google Scholar 

  50. 50.

    Sobin, L. H. & Fleming, I. D. Review of TNM classification of malignant tumors, fifth edition. Cancer 80, 1803–1804 (1997).

    CAS  Article  Google Scholar 

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This study was supported by a research project promoted by the Grants-in-Aid for Scientific Research promoted by the Ministry of Education, Culture, Sports, Science, and Technology in Japan (17H04365, 18K19950, and 18H04707, received by A.K.) and the Japan Agency for Medical Research and Development (AMED-PRIME, received by A.K.).

Author information




A.K. designed the study, conducted the experiments, and wrote the paper, with assistance from A.S. and T.O. Y.H. conducted the analyses. S.K. and K.K generated some of the viral vectors and rats. R.K. and Y.Y. generated some of the rats. T.S. and K.I. generated the NaChBac mutant and conducted the in vitro electrophysiological experiments.

Corresponding author

Correspondence to Atsunori Kamiya.

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Competing interests

The authors declare no competing interests except the patent (patent applicant: Asunori Kamiya; name of inventor: Asunori Kamiya; application number: PCT/JP2017/25468; status of application: international migration; specific aspect of manuscript covered in patent application: the genetic engineering of local nerves including tumoral autonomic nerves; the concept of genetic engineering of local nerves for treatment of variable diseases including cancers; the viral vectors and their constructions to control, stimulate and delete sympathetic nerves; and the viral vectors and their constructions to control, stimulate and delete parasympathetic nerves).

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Peer review information: Nature Neuroscience thanks Paul Frenette and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Kamiya, A., Hayama, Y., Kato, S. et al. Genetic manipulation of autonomic nerve fiber innervation and activity and its effect on breast cancer progression. Nat Neurosci 22, 1289–1305 (2019).

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