Background & Aims
Oral cancer patients suffer pain at the site of the cancer. Nerves contribute to the pathogenesis of oral squamous cell carcinoma (OSCC) (2). They are recruited and co-opted by cancers (3, 4). Nerve density correlates with more aggressive disease, including for cancers of the head and neck (5-7). The tongue is innervated by sensory, sympathetic and parasympathetic neurons carried by four nerves, the lingual nerve, chorda tympani, glossopharyngeal nerve and postganglionic autonomic nerves. The aims of this research are to map the connectome between the OSCC tumor and the ganglia, identify the involved neuronal cell types and assess variations associated with the site on the tongue.
Methods
Oral cancer tongue allograft mouse cancer models were generated by injecting 4MOSC2 (more aggressive) and 4MOSC1 oral cancer cells in a diluent of a 1:1 mixture of saline and matrigel into the anterior tongue (right and left) of C57BL/6 mice, injection of a 1:1 saline-matrigel only was done for the control group. Neurons innervating the cancers were identified by injecting the neural tracer 4% fluorogold (no transcytosis) into the cancers 7 days prior to harvesting trigeminal ganglia from the mice. The ganglia were fixed, OCT embedded, sectioned and the sections stained with antibodies to neuronal markers. Imaging of the whole trigeminal ganglion was done using the Zeiss 800 confocal microscope, 20x objective (z-stack, tile). Cell were counted with Image J counter. Two sections per slide were counted for each animal
Results
Retrograde labeling was present predominantly in select neuronal cell bodies (confirmed by NEUN IHC) belonging to the mandibular (V3) branch of the trigeminal ganglion with some labeling of cell bodies in the ophthalmic (V1) and maxillary (V2) clusters. Evidence of oral cancer associated reprogramming of TG neurons innervating 4MOSC1 and 4MOSC2 tongue tumor allografts was obtained that is consistent with and/or extended previous reports, including increased neuronal expression of TRPA1 (P=0.018), TRPV1 (P=0.048) and the voltage gated channel Nav1.8, encoded by Scn10a (P=0.002) in the cancer group (8-11), as well as increased expression of OCT4 (encoded by Pou5f1) and KLF4 (12). Differential upregulation of developmental reprogramming genes and pain mediators is being assessed in trigeminal neurons and satellite glial cells with respect to aggressiveness of the cancer cell lines.
Conclusions
Recruitment and reprogramming of lingual nerves by OSSC tumors at both the right and left anterior fields of the tongue of the mice was confirmed. Crosstalk of the lingual nerves (V3) with certain neuronal cell types of the ophthalmic (V1) and maxillary (V2) clusters exists and may be relevant to OSSC tumor biology, progression and pain.
Relevance to Patient care: sympathectomy in a carcinogen-induced mouse model of OSCC
and surgical transection of nerves in lip tumor in the clinical setting respectively led to OSCC
regression and symptomatic control (reduced ulceration and pain), thus peripheral nerves
contribute to oral tumor’s biology and pain. The present study provides insight into the identity
of the peripheral nerve subtypes recruited by the oral tumor; the possible mechanisms involved
and how these neuronal cell types contribute to the tumor progression, nociceptive behavior
and metastasis. An understanding will provide a noninvasive therapeutic way of controlling oral
cancer pain and tumor progression
References
- Pires FR, Ramos AB, Oliveira JBCd, Tavares AS, Luz PSRd, Santos TCRBd. Oral squamous cell carcinoma: clinicopathological features from 346 cases from a single Oral Pathology service during an 8-year period. Journal of Applied Oral Science. 2013; 21(5): 460- 7.
- Zahalka AH, Frenette PS. Nerves in cancer. Nature Reviews Cancer. 2020; 20(3):143-57.
- Zahalka AH, Arnal-Estapé A, Maryanovich M, Nakahara F, Cruz CD, Finley LWS, et al.
Adrenergic nerves activate an angio-metabolic switch in prostate cancer. Science. 2017; 358(6361): 321-6.
- Renz BW, Takahashi R, Tanaka T, Macchini M, Hayakawa Y, Dantes Z, et al. β2 Adrenergic-Neurotrophin Feedforward Loop Promotes Pancreatic Cancer. Cancer Cell. 2018; 33(1):75-90.e7.
- Albo D, Akay CL, Marshall CL, Wilks JA, Verstovsek G, Liu H, et al. Neurogenesis in colorectal cancer is a marker of aggressive tumor behavior and poor outcomes. Cancer. 2011; 117(21): 4834-45.
6 Huang D, Su S, Cui X, Shen X, Zeng Y, Wu W, et al. Nerve Fibers in Breast Cancer Tissues Indicate Aggressive Tumor Progression. Medicine. 2014; 93(27): e172.
- Ayala GE, Dai H, Powell M, Li R, Ding Y, Wheeler TM, et al. Cancer-Related Axonogenesis and Neurogenesis in Prostate Cancer. Clinical Cancer Research. 2008; 14(23): 7593-603.
- Raju B, Haug SR, Ibrahim SO & Heyeraas KJ Sympathectomy decreases size and invasiveness of tongue cancer in rats. Neuroscience 149, 715–725, doi:10.1016/j.neuroscience.2007.07.048 (2007). [PubMed: 17916410]
- Scheff NN, Wall IM, Nicholson S, Williams H, Chen E, Tu NH et al. Oral cancer induced TRPV1 sensitization is mediated by PAR2signaling in primary afferent neurons innervating the cancer microenvironment. Sci Rep. 2022;12(1):4121. doi: 10.1038/s41598-022-08005-6. PMID: 35260737; PMCID: PMC8904826.
- Ruparel S, Bendele M, Wallace A, Green D. Released lipids regulate transient receptor potential channel (TRP)-dependent oral cancer pain. Mol Pain. 2015: 11:30. doi: 10.1186/s12990-015-0016-3. PMID: 26007300; PMCID: PMC4456056.
- Gonzales CB, De La Chapa JJ, Patwardhan AM, Hargreaves KM. Oral Cancer Pain Includes Thermal Allodynia That May Be Attenuated by Chronic Alcohol Consumption. Pharmaceuticals (Basel). 2023;16(4):518. doi: 10.3390/ph16040518. PMID: 37111275; PMCID: PMC10142169.
- Amit M, Takahashi H, Dragomir MP, Lindemann A, Gleber-Netto FO, Pickering CR et al. Loss of p53 drives neuron reprogramming in head and neck cancer. Nature. 2020 ;578(7795):449-454. doi: 10.1038/s41586-020-1996-3. Epub 2020 Feb 12. PMID: 32051587; PMCID: PMC9723538.
Presenting Author
Alex Agyemang
Poster Authors
Alex Agyemang
PhD
Translational Research Center, New York University College of Dentistry, New York,
Lead Author
Donna Albertson
Translational Research Center, New York University College of Dentistry, New York,
Lead Author
Topics
- Models: Oral/craniofacial