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

Abstract

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.

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Acknowledgements

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.).

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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). https://doi.org/10.1038/s41593-019-0430-3

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