Abstract
Post-surgical pain remains a widespread problem reducing quality of life. The present study investigates the initial molecular changes underlying nociceptive sensitization through longitudinal, temporal sampling at the surgical wound edge. Using RNA-Seq and multiplex fluorescence in situ hybridization, we examined the most significant genes induced by tissue injury including those coding for the secreted factors interleukin 6, oncostatin M and leukemia inhibitory factor, and localized these induction events to several cutaneous structures including the epidermis, vascular endothelia, hair follicles, and sweat glands. Our data also demonstrate the receptors for these key secreted factors are expressed by dorsal root ganglion neurons, indicating long-range signaling from damaged skin to spinal cord, thereby leading to pain. This study provides a novel understanding of tissue structures and the molecular interactome activated following tissue injury by elucidating the inflammatory and tissue repair transcriptional milieu induced by surgery in human skin excision biopsies.
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Data availability
Sequencing data were deposited in public databases (Sequence Read Archive, BioProject# PRJNA1154260; dbGaP: phs003890) alongside sample-level patient-reported outcomes and data dictionaries. Large supplementary data tables (Supplementary Tables 1–5) are also available on figshare (https://doi.org/10.6084/m9.figshare.27384174). Source data underlying all graphs in the manuscript can be found in Supplementary Data 1.
References
Glare, P., Aubrey, K. R. & Myles, P. S. Transition from acute to chronic pain after surgery. Lancet 393, 1537–1546 (2019).
Neuman, M. D., Bateman, B. T. & Wunsch, H. Inappropriate opioid prescription after surgery. Lancet 393, 1547–1557 (2019).
Brennan, T. J. Pathophysiology of postoperative pain. Pain 152, S33–S40 (2011).
Meyer, R. A., Ringkamp, M., Campbell, J. N. & Raja, S. N. Peripheral mechanisms of cutaneous nociception. In Wall and Melzack’s Textbook of Pain (eds McMahon, S. B. & Koltzenburg, M.) 3–34 (Elsevier, London, 2006).
Goto, T. et al. Longitudinal peripheral tissue RNA-Seq transcriptomic profiling, hyperalgesia, and wound healing in the rat plantar surgical incision model. FASEB J. 35, e21852 (2021).
Gold, M. S. & Gebhart, G. F. Nociceptor sensitization in pain pathogenesis. Nat. Med. 16, 1248–1257 (2010).
Cook, S. P., Vulchanova, L., Hargreaves, K. M., Elde, R. & McCleskey, E. W. Distinct ATP receptors on pain-sensing and stretch-sensing neurons. Nature 387, 505–508 (1997).
Goto, T. et al. Longitudinal transcriptomic profiling in carrageenan-induced rat hind paw peripheral inflammation and hyperalgesia reveals progressive recruitment of innate immune system components. J. Pain. 22, 322–343 (2021).
Brennan, T. J., Vandermeulen, E. P. & Gebhart, G. F. Characterization of a rat model of incisional pain. Pain 64, 493–502 (1996).
Cleeland, C. S. & Ryan, K. M. Pain assessment: global use of the Brief Pain Inventory. Ann. Acad. Med. Singap. 23, 129–138 (1994).
Melzack, R. The short-form McGill pain questionnaire. Pain 30, 191–197 (1987).
LaPaglia, D. M. et al. RNA-Seq investigations of human post-mortem trigeminal ganglia. Cephalalgia 38, 912–932 (2018).
Wang, C. et al. The concordance between RNA-seq and microarray data depends on chemical treatment and transcript abundance. Nat. Biotechnol. 32, 926–932 (2014).
Sapio, M. R., Goswami, S. C., Gross, J. R., Mannes, A. J. & Iadarola, M. J. Transcriptomic analyses of genes and tissues in inherited sensory neuropathies. Exp. Neurol. 283, 375–395 (2016).
Lessard, J. C. et al. Keratin 16 regulates innate immunity in response to epidermal barrier breach. Proc. Natl. Acad. Sci. USA 110, 19537–19542 (2013).
Lin, P. H. et al. Zinc in wound healing modulation. Nutrients 10, https://doi.org/10.3390/nu10010016 (2017).
Wolf, C. L., Pruett, C., Lighter, D. & Jorcyk, C. L. The clinical relevance of OSM in inflammatory diseases: a comprehensive review. Front. Immunol. 14, 1239732 (2023).
Mwirigi, J. et al. Oncostatin M induces nociceptive signaling in human dorsal root ganglia. J. Pain. 24, 16–16 (2023).
Tseng, P. Y. & Hoon, M. A. Oncostatin M can sensitize sensory neurons in inflammatory pruritus. Sci. Transl. Med. 13, eabe3037 (2021).
Ghitani, N. et al. Specialized mechanosensory nociceptors mediating rapid responses to hair pull. Neuron 95, 944–954.e944 (2017).
von Buchholtz, L. J. et al. Decoding cellular mechanisms for mechanosensory discrimination. Neuron 109, 285–298.e285 (2021).
Yu, H. et al. Leveraging deep single-soma RNA sequencing to explore the neural basis of human somatosensation. Nature Neurosci. 27, 2326–2340 (2024).
Caterina, M. J. et al. The capsaicin receptor: a heat-activated ion channel in the pain pathway. Nature 389, 816–824 (1997).
Ma, W. et al. Anatomical analysis of transient potential vanilloid receptor 1 (Trpv1+) and Mu-opioid receptor (Oprm1+) co-expression in rat dorsal root ganglion neurons. Front. Mol. Neurosci. 15, 926596 (2022).
Sapio, M. R. et al. Expression pattern analysis and characterization of the hereditary sensory and autonomic neuropathy 2 A (HSAN2A) gene with no lysine kinase (WNK1) in human dorsal root ganglion. Exp. Neurol. 370, 114552 (2023).
Sapio, M. R. et al. Analgesic candidate adenosine A 3 receptors are expressed by perineuronal peripheral macrophages in human dorsal root ganglion and spinal cord microglia. Pain 165, 2323–2343 (2024).
Tavares-Ferreira, D. et al. Spatial transcriptomics of dorsal root ganglia identifies molecular signatures of human nociceptors. Sci. Transl. Med. 14, eabj8186 (2022).
Staedtler, E. S. et al. The mu-opioid receptor differentiates two distinct human nociceptive populations relevant to clinical pain. Cell Rep. Med. 5, 101788 (2024).
Rittie, L., Sachs, D. L., Orringer, J. S., Voorhees, J. J. & Fisher, G. J. Eccrine sweat glands are major contributors to reepithelialization of human wounds. Am. J. Pathol. 182, 163–171 (2013).
Futagami, A., Ishizaki, M., Fukuda, Y., Kawana, S. & Yamanaka, N. Wound healing involves induction of cyclooxygenase-2 expression in rat skin. Lab. Investig. 82, 1503–1513 (2002).
Loynes, C. A. et al. PGE(2) production at sites of tissue injury promotes an anti-inflammatory neutrophil phenotype and determines the outcome of inflammation resolution in vivo. Sci. Adv. 4, eaar8320 (2018).
De Jongh, R. F. et al. The role of interleukin-6 in nociception and pain. Anesth. Analg. 96, 1096–1103 (2003).
Wei, X. H. et al. The up-regulation of IL-6 in DRG and spinal dorsal horn contributes to neuropathic pain following L5 ventral root transection. Exp. Neurol. 241, 159–168 (2013).
Sole-Boldo, L. et al. Single-cell transcriptomes of the human skin reveal age-related loss of fibroblast priming. Commun. Biol. 3, 188 (2020).
Raithel, S. J., Sapio, M. R., LaPaglia, D. M., Iadarola, M. J. & Mannes, A. J. Transcriptional changes in dorsal spinal cord persist after surgical incision despite preemptive analgesia with peripheral resiniferatoxin. Anesthesiology 128, 620–635 (2018).
Spofford, C. M. & Brennan, T. J. Gene expression in skin, muscle, and dorsal root ganglion after plantar incision in the rat. Anesthesiology 117, 161–172 (2012).
Segelcke, D. et al. Phenotype- and species-specific skin proteomic signatures for incision-induced pain in humans and mice. Br. J. Anaesth. 130, 331–342 (2023).
Brummett, C. M. et al. New persistent opioid use after minor and major surgical procedures in US adults. JAMA Surg. 152, e170504 (2017).
Alam, A. et al. Long-term analgesic use after low-risk surgery: a retrospective cohort study. Arch. Intern. Med. 172, 425–430 (2012).
Sapio, M. R. et al. The persistent pain transcriptome: identification of cells and molecules activated by hyperalgesia. J. Pain. 22, 1146–1179 (2021).
Parisien, M. et al. Acute inflammatory response via neutrophil activation protects against the development of chronic pain. Sci. Transl. Med. 14, eabj9954 (2022).
Huerta, M. Á. et al. The role of neutrophils in pain: systematic review and meta-analysis of animal studies. Pain 166, 1230–1249 (9900).
Mitchell, M. E. et al. Interleukin-6 induces nascent protein synthesis in human dorsal root ganglion nociceptors primarily via MNK-eIF4E signaling. Neurobiol. Pain 16, 100159 (2024).
Jawa, R. S., Anillo, S., Huntoon, K., Baumann, H. & Kulaylat, M. Analytic review: interleukin-6 in surgery, trauma, and critical care: part I: basic science. J. Intensive Care Med. 26, 3–12 (2011).
Morellini, N. M. et al. Exogenous metallothionein-IIA promotes accelerated healing after a burn wound. Wound Repair Regen. 16, 682–690 (2008).
Das, A. et al. Oncostatin M improves cutaneous wound re-epithelialization and is deficient under diabetic conditions. J. Investig. Dermatol. 142, 679–691.e673 (2022).
Langeslag, M. et al. Oncostatin M induces heat hypersensitivity by gp130-dependent sensitization of TRPV1 in sensory neurons. Mol. Pain. 7, 102 (2011).
Spofford, C. M., Mohan, S., Kang, S., Jang, J. H. & Brennan, T. J. Evaluation of leukemia inhibitory factor (LIF) in a rat model of postoperative pain. J. Pain. 12, 819–832 (2011).
Banner, L. R., Patterson, P. H., Allchorne, A., Poole, S. & Woolf, C. J. Leukemia inhibitory factor is an anti-inflammatory and analgesic cytokine. J. Neurosci. 18, 5456–5462 (1998).
Smith, A. F., Plumb, A. N., Berardi, G. & Sluka, K. A. Sex differences in the transition to chronic pain. J. Clin. Investig. 135, https://doi.org/10.1172/JCI191931 (2025).
Schreiber, K. L. et al. Prediction of persistent pain severity and impact 12 months after breast surgery using comprehensive preoperative assessment of biopsychosocial pain modulators. Ann. Surg. Oncol. 28, 5015–5038 (2021).
Dworkin, R. H. et al. Core outcome measures for chronic pain clinical trials: IMMPACT recommendations. Pain 113, 9–19 (2005).
Chen, E. Y. et al. Enrichr: interactive and collaborative HTML5 gene list enrichment analysis tool. BMC Bioinform. 14, 128 (2013).
Sapio, M. R. et al. Efficient removal of naturally-occurring lipofuscin autofluorescence in human nervous tissue using high-intensity white light. J. Pain 30, 105359 (2025).
Maric, D. et al. Whole-brain tissue mapping toolkit using large-scale highly multiplexed immunofluorescence imaging and deep neural networks. Nat. Commun. 12, 1550 (2021).
Sapio, M. R. et al. Comparative analysis of dorsal root, nodose and sympathetic ganglia for the development of new analgesics. Front. Neurosci. 14, 615362 (2020).
Harding, S. D. et al. The IUPHAR/BPS guide to PHARMACOLOGY in 2024. Nucleic Acids Res. 52, D1438–D1449 (2024).
Binder, J. X. et al. COMPARTMENTS: unification and visualization of protein subcellular localization evidence. Database 2014, bau012 (2014).
Stelzer, G. et al. The GeneCards Suite: from gene data mining to disease genome sequence analyses. Curr. Protoc. Bioinform. 54, 1 30 31–31 30 33 (2016).
UniProt, C. UniProt: the universal protein knowledgebase in 2021. Nucleic Acids Res. 49, D480–D489 (2021).
Ray, P. R. et al. A pharmacological interactome between COVID-19 patient samples and human sensory neurons reveals potential drivers of neurogenic pulmonary dysfunction. Brain Behav. Immun. 89, 559–568 (2020).
Acknowledgements
This study was supported by the Intramural Research Program of the National Institutes of Health Clinical Center (ZIACL090034-09, ZIACL090035-08, ZIACL0033-09 to A.J.M.) and of the National Institute of Neurological Disorders and Stroke. Supplementary funding was provided by the Office of Behavioral and Social Science Research and from a Bench to Bedside Grant from the NIH. T.G. was the recipient of a JSPS Overseas Research Fellowship from April 2018 to March 2020 from the Japan Society for the Promotion of Science. This work was funded by the National institutes of Health. The contributions of the NIH author(s) are considered Works of the United States Government. The findings and conclusions presented in this paper are those of the author(s) and do not necessarily reflect the views of the NIH or the U.S. Department of Health and Human Services.
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This study was conceptualized and designed by M.R.S., with input from M.J.I. and A.J.M. M.R.S. led the project from inception through completion, including protocol development, data acquisition, analysis, and manuscript preparation. M.R.S. wrote the first draft of the manuscript, with input and editing from A.J.M., E.L., T.G., and D.M. All authors reviewed and approved the final version. Clinical protocol development was led by M.R.S. with supervision and guidance from T.S.W. and A.J.M., with additional consultation from M.J.I., J.L.D., and D.S.S. Surgeries and tissue excision were performed by A.M.B., J.L.D., J.M.H., and D.S.S. Sample collection was coordinated by M.R.S. and A.F.D., with subsequent processing by TG. Histological assessments were performed by S.T. In situ hybridization experiments were conducted by E.L. and A.P.M., with additional consultation regarding interpretation from S.T. and D.M. Microscopy was performed by E.L. and A.P.M., with supervision and experimental design consultation from D.M. Visualizations and data figures were designed and generated by M.R.S. and E.L. Formal data analysis was led by M.R.S. with contributions from T.G. Project supervision and coordination were carried out by M.R.S. with input from A.J.M. and T.S.W. Funding support was provided by A.J.M.
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Communications Biology thanks Daniel Segelcke, Nynke J. van Den Hoogen, and the other anonymous reviewer for their contribution to the peer review of this work. Primary handling editors: Joao Valente.
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Sapio, M.R., Li, E., Domenichiello, A.F. et al. Longitudinal human transcriptomic and spatial gene profiling at the incisional edge during long surgical procedures. Commun Biol (2025). https://doi.org/10.1038/s42003-025-09366-0
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DOI: https://doi.org/10.1038/s42003-025-09366-0
