CXCL10 Promotes Spinal Macrophage Recruitment via the JAK/STAT3 Pathway to Induce Pain in Experimental Autoimmune Prostatitis

Abstract

The aim is to explore the mechanisms underlying pain development in chronic prostatitis and identify therapeutic targets for pain management in patients with chronic prostatitis. RNA sequence of the spinal cord dorsal horns and proteomic analysis of spinal macrophages of experimental autoimmune prostatitis (EAP) mice were conducted to identify pain-related genes, proteins and signalling pathways. The clodronate liposome, CXCR3 and P-STAT3 inhibitors, NGF antibody and cromolyn sodium were used to investigate the roles of the CXCL10/CXCR3, JAK/STAT3 and NGF/TrKA pathways in spinal macrophage recruitment and pain response. Finally, prostate tissues from benign prostate hyperplasia (BPH) patients were collected to validate the aforementioned results. Neuron and astrocyte-derived CXCL10 was associated with spinal macrophage recruitment, and CXCL10/CXCR3 axis could regulate the chemotaxis of macrophage to the spinal cord in EAP mice. Results of proteomic analysis found that CXCL10 could regulate the JAK/STAT3 pathway to mediate neuroinflammation in EAP, which was validated in vivo and in vitro experiments. The number of mast cells and expressions of NGF, TrKA and PGP9.5 increased in the prostates of EAP mice and BPH patients, and targeting NGF could reduce spinal macrophage recruitment and pain response. NGF was the triggering factor to induce chemotaxis of spinal macrophages and neuroinflammation, and the CXCL10/CXCR3 axis and JAK/STAT3 pathway was involved in spinal macrophage recruitment and infiltration, which provided therapeutic targets for pain management.

Keywords

chronic prostatitis / CXCL10 / nerve growth factor / pain / spinal macrophages / STAT3

References

[1] A. S. Polackwich and D. A. Shoskes, “Chronic Prostatitis/Chronic Pelvic Pain Syndrome: A Review of Evaluation and Therapy,” Prostate Cancer and Prostatic Diseases 19 (2016): 132-138.

[2] A. C. Strauss and J. D. Dimitrakov, “New Treatments for Chronic Prostatitis/Chronic Pelvic Pain Syndrome,” Nature Reviews. Urology 7 (2010): 127-135.

[3] G. G. Page, M. R. Opp, and S. L. Kozachik, “Reduced Sleep, Stress Responsivity, and Female Sex Contribute to Persistent Inflammation-Induced Mechanical Hypersensitivity in Rats,” Brain, Behavior, and Immunity 40 (2014): 244-251.

[4] X. Wu, K. Cheng, C. Xu, et al., “Mechanism of Acupuncture and Moxibustion on Chronic Prostatitis/Chronic Pelvic Pain Syndrome: A Narrative Review of Animal Studies,” Pain Research & Management 2021 (2021): 2678242.

[5] J. Zhang, C. Liang, X. Shang, and H. Li, “Chronic Prostatitis/Chronic Pelvic Pain Syndrome: A Disease or Symptom? Current Perspectives on Diagnosis, Treatment, and Prognosis,” American Journal of Men's Health 14 (2020): 1557988320903200.

[6] R. R. Ji, Z. Z. Xu, and Y. J. Gao, “Emerging Targets in Neuroinflammation-Driven Chronic Pain,” Nature Reviews. Drug Discovery 13 (2014): 533-548.

[7] D. N. Xanthos and J. Sandkühler, “Neurogenic Neuroinflammation: Inflammatory CNS Reactions in Response to Neuronal Activity,” Nature Reviews. Neuroscience 15 (2014): 43-53.

[8] R. R. Ji, A. Chamessian, and Y. Q. Zhang, “Pain Regulation by Non-neuronal Cells and Inflammation,” Science (New York, N.Y.) 354 (2016): 572-577.

[9] Y. Lao, Z. Li, Y. Bai, et al., “Glial Cells of the Central Nervous System: A Potential Target in Chronic Prostatitis/Chronic Pelvic Pain Syndrome,” Pain Research & Management 2023 (2023): 2061632.

[10] H. Zhang, R. Gu, J. Luo, C. Zhong, and J. Pan, “Involvement of NOTCH1-Mediated Microglia Activation in Neuromodulation of Chronic Prostatitis-Related Pain,” In Vivo (Athens, Greece) 38 (2024): 691-698.

[11] M. Zelenka, M. Schäfers, and C. Sommer, “Intraneural Injection of Interleukin-1beta and Tumor Necrosis Factor-Alpha Into Rat Sciatic Nerve at Physiological Doses Induces Signs of Neuropathic Pain,” Pain 116 (2005): 257-263.

[12] M. W. Salter and S. Beggs, “Sublime Microglia: Expanding Roles for the Guardians of the CNS,” Cell 158 (2014): 15-24.

[13] M. Tsuda, K. Koga, T. Chen, and M. Zhuo, “Neuronal and Microglial Mechanisms for Neuropathic Pain in the Spinal Dorsal Horn and Anterior Cingulate Cortex,” Journal of Neurochemistry 141 (2017): 486-498.

[14] X. Yu, H. Liu, K. A. Hamel, et al., “Dorsal Root Ganglion Macrophages Contribute to Both the Initiation and Persistence of Neuropathic Pain,” Nature Communications 11 (2020): 264.

[15] M. Tsuda, Y. Shigemoto-Mogami, S. Koizumi, et al., “P2X4 Receptors Induced in Spinal Microglia Gate Tactile Allodynia After Nerve Injury,” Nature 424 (2003): 778-783.

[16] H. Kikutani and S. Makino, “The Murine Autoimmune Diabetes Model: NOD and Related Strains,” Advances in Immunology 51 (1992): 285-322.

[17] V. E. Rivero, C. Cailleau, M. Depiante-Depaoli, C. M. Riera, and C. Carnaud, “Non-Obese Diabetic (NOD) Mice Are Genetically Susceptible to Experimental Autoimmune Prostatitis (EAP),” Journal of Autoimmunity 11 (1998): 603-610.

[18] W. M. Ridgway, “Dissecting Genetic Control of Autoimmunity in NOD Congenic Mice,” Immunologic Research 36 (2006): 189-195.

[19] M. L. Breser, R. D. Motrich, L. R. Sanchez, J. P. Mackern-Oberti, and V. E. Rivero, “Expression of CXCR3 on Specific T Cells Is Essential for Homing to the Prostate Gland in an Experimental Model of Chronic Prostatitis/Chronic Pelvic Pain Syndrome,” Journal of Immunology 190 (2013): 3121-3133.

[20] Y. F. Kong, W. L. Sha, X. B. Wu, L. X. Zhao, L. J. Ma, and Y. J. Gao, “CXCL10/CXCR3 Signaling in the DRG Exacerbates Neuropathic Pain in Mice,” Neuroscience Bulletin 37 (2021): 339-352.

[21] Y. Y. Ju, M. Jiang, F. Xu, et al., “CXCL10 and CXCR3 in the Trigeminal Ganglion Contribute to Trigeminal Neuropathic Pain in Mice,” Journal of Pain Research 14 (2021): 41-51.

[22] Y. Chen, D. Yin, B. Fan, et al., “Chemokine CXCL10/CXCR3 Signaling Contributes to Neuropathic Pain in Spinal Cord and Dorsal Root Ganglia After Chronic Constriction Injury in Rats,” Neuroscience Letters 694 (2019): 20-28.

[23] Z. Liu, S. F. Murphy, L. Wong, A. J. Schaeffer, and P. Thumbikat, “Neuronal/Astrocytic Expression of Chemokine (C-C Motif) Ligand 2 Is Associated With Monocyte/Macrophage Recruitment in Male Chronic Pelvic Pain,” Pain 161 (2020): 2581-2591.

[24] X. Hua, S. Ge, M. Zhang, et al., “Pathogenic Roles of CXCL10 in Experimental Autoimmune Prostatitis by Modulating Macrophage Chemotaxis and Cytokine Secretion,” Frontiers in Immunology 12 (2021): 706027.

[25] L. G. Zhang, J. Chen, J. L. Meng, et al., “Effect of Alcohol on Chronic Pelvic Pain and Prostatic Inflammation in a Mouse Model of Experimental Autoimmune Prostatitis,” Prostate 79 (2019): 1439-1449.

[26] M. T. Brown, F. T. Murphy, D. M. Radin, I. Davignon, M. D. Smith, and C. R. West, “Tanezumab Reduces Osteoarthritic Hip Pain: Results of a Randomized, Double-Blind, Placebo-Controlled Phase III Trial,” Arthritis and Rheumatism 65 (2013): 1795-1803.

[27] J. D. Done, C. N. Rudick, M. L. Quick, A. J. Schaeffer, and P. Thumbikat, “Role of Mast Cells in Male Chronic Pelvic Pain,” Journal of Urology 187 (2012): 1473-1482.

[28] L. Chen, Y. Liu, S. Yue, et al., “P2X7R Modulates NEK7-NLRP3 Interaction to Exacerbate Experimental Autoimmune Prostatitis via GSDMD-Mediated Prostate Epithelial Cell Pyroptosis,” International Journal of Biological Sciences 20 (2024): 3393-3411.

[29] X. Hua, J. Zhang, S. Ge, et al., “CXCR3 Antagonist AMG487 Ameliorates Experimental Autoimmune Prostatitis by Diminishing Th1 Cell Differentiation and Inhibiting Macrophage M1 Phenotypic Activation,” Prostate 82 (2022): 1223-1236.

[30] J. Zhang, Y. Liu, H. Chen, et al., “MyD88 in Hepatic Stellate Cells Enhances Liver Fibrosis via Promoting Macrophage M1 Polarization,” Cell Death & Disease 13 (2022): 411.

[31] S. Chen, Y. Zhou, Y. Chen, and J. Gu, “Fastp: An Ultra-Fast All-in-One FASTQ Preprocessor,” Bioinformatics 34 (2018): i884-i890.

[32] D. Kim, B. Langmead, and S. L. Salzberg, “HISAT: A Fast Spliced Aligner With Low Memory Requirements,” Nature Methods 12 (2015): 357-360.

[33] A. Roberts, C. Trapnell, J. Donaghey, J. L. Rinn, and L. Pachter, “Improving RNA-Seq Expression Estimates by Correcting for Fragment Bias,” Genome Biology 12 (2011): R22.

[34] S. Anders, P. T. Pyl, and W. Huber, “HTSeq—A Python Framework to Work With High-Throughput Sequencing Data,” Bioinformatics 31 (2015): 166-169.

[35] The Gene Ontology Consortium, “The Gene Ontology Resource: 20 Years and Still Going Strong,” Nucleic Acids Research 47 (2019): D330-D338.

[36] M. Kanehisa, M. Araki, S. Goto, et al., “KEGG for Linking Genomes to Life and the Environment,” Nucleic Acids Research 36 (2008): D480-D484.

[37] A. Subramanian, P. Tamayo, V. K. Mootha, et al., “Gene Set Enrichment Analysis: A Knowledge-Based Approach for Interpreting Genome-Wide Expression Profiles,” Proceedings of the National Academy of Sciences of the United States of America 102 (2005): 15545-15550.

[38] P. Langfelder and S. Horvath, “WGCNA: An R Package for Weighted Correlation Network Analysis,” BMC Bioinformatics 9 (2008): 559.

[39] Y. Li, R. M. Ritzel, N. Khan, et al., “Delayed Microglial Depletion After Spinal Cord Injury Reduces Chronic Inflammation and Neurodegeneration in the Brain and Improves Neurological Recovery in Male Mice,” Theranostics 10 (2020): 11376-11403.

[40] J. C. Nickel, L. D. True, J. N. Krieger, R. E. Berger, A. H. Boag, and I. D. Young, “Consensus Development of a Histopathological Classification System for Chronic Prostatic Inflammation,” BJU International 87 (2001): 797-805.

[41] G. C. Deng, M. Lu, Y. Y. Zhao, Y. Yuan, and G. Chen, “Activated Spinal Astrocytes Contribute to the Later Phase of Carrageenan-Induced Prostatitis Pain,” Journal of Neuroinflammation 16 (2019): 189.

[42] L. Wong, J. D. Done, A. J. Schaeffer, and P. Thumbikat, “Experimental Autoimmune Prostatitis Induces Microglial Activation in the Spinal Cord,” Prostate 75 (2015): 50-59.

[43] M. Ishigooka, D. H. Zermann, R. Doggweiler, and R. A. Schmidt, “Similarity of Distributions of Spinal c-Fos and Plasma Extravasation After Acute Chemical Irritation of the Bladder and the Prostate,” Journal of Urology 164 (2000): 1751-1756.

[44] S. I. Grivennikov and M. Karin, “Dangerous Liaisons: STAT3 and NF-kappaB Collaboration and Crosstalk in Cancer,” Cytokine & Growth Factor Reviews 21 (2010): 11-19.

[45] G. He and M. Karin, “NF-κB and STAT3—Key Players in Liver Inflammation and Cancer,” Cell Research 21 (2011): 159-168.

[46] B. Nicholson and S. Verma, “Comorbidities in Chronic Neuropathic Pain,” Pain Medicine (Malden, Mass.) 5, no. Suppl 1 (2004): S9-S27.

[47] H. Zhang, L. Liu, Z. Yang, et al., “P2X7 Receptor Mediates Activation of Microglial Cells in Prostate of Chemically Irritated Rats,” International Brazilian Journal of Urology 39 (2013): 276-285.

[48] H. Zhang, L. M. Liu, G. S. Lu, et al., “Correlation Between Activation of L5-S2 Spinal Cord Astrocytes and Effect of Substance P in Chronic Prostatitis Pain,” Zhonghua Nan Ke Xue = National Journal of Andrology 15 (2009): 1021-1027.

[49] C. Zhang and D. Li, “Effects of Electroacupuncture on Alleviating Prostatodynia and Inflammation in Rats With Chronic Nonbacterial Prostatitis,” Journal of Pain Research 14 (2021): 2757-2765.

[50] S. Nencini, M. Ringuet, D. H. Kim, Y. J. Chen, C. Greenhill, and J. J. Ivanusic, “Mechanisms of Nerve Growth Factor Signaling in Bone Nociceptors and in an Animal Model of Inflammatory Bone Pain,” Molecular Pain 13 (2017): 1744806917697011.

[51] S. Pezet and S. B. McMahon, “Neurotrophins: Mediators and Modulators of Pain,” Annual Review of Neuroscience 29 (2006): 507-538.

[52] L. Aloe, M. A. Tuveri, U. Carcassi, and R. Levi-Montalcini, “Nerve Growth Factor in the Synovial Fluid of Patients With Chronic Arthritis,” Arthritis and Rheumatism 35 (1992): 351-355.

[53] M. Ueda, M. Hirose, N. Takei, et al., “Nerve Growth Factor Induces Systemic Hyperalgesia After Thoracic Burn Injury in the Rat,” Neuroscience Letters 328 (2002): 97-100.

[54] Q. P. Ma and C. J. Woolf, “The Progressive Tactile Hyperalgesia Induced by Peripheral Inflammation Is Nerve Growth Factor Dependent,” Neuroreport 8 (1997): 807-810.

[55] S. Govoni, P. Fantucci, N. Marchesi, et al., “N-Acetylcysteine Antagonizes NGF Activation of TrkA Through Disulfide Bridge Interaction, an Effect Which May Contribute to Its Analgesic Activity,” International Journal of Molecular Sciences 25 (2023): 206.

[56] F. Denk, D. L. Bennett, and S. B. McMahon, “Nerve Growth Factor and Pain Mechanisms,” Annual Review of Neuroscience 40 (2017): 307-325.

[57] N. Khan and M. T. Smith, “Neurotrophins and Neuropathic Pain: Role in Pathobiology,” Molecules (Basel, Switzerland) 20 (2015): 10657-10688.

[58] M. Enomoto, P. W. Mantyh, J. Murrell, J. F. Innes, and B. D. X. Lascelles, “Anti-Nerve Growth Factor Monoclonal Antibodies for the Control of Pain in Dogs and Cats,” Veterinary Record 184 (2019): 23.

[59] M. P. Webb, E. M. Helander, B. L. Menard, R. D. Urman, and A. D. Kaye, “Tanezumab: A Selective Humanized mAb for Chronic Lower Back Pain,” Therapeutics and Clinical Risk Management 14 (2018): 361-367.

[60] A. J. Poole, L. Frigotto, M. E. Smith, et al., “A C-Terminal Cysteine Residue Is Required for Peptide-Based Inhibition of the NGF/TrkA Interaction at nM Concentrations: Implications for Peptide-Based Analgesics,” Scientific Reports 9 (2019): 930.

[61] J. N. Katz, “Tanezumab for Painful Osteoarthritis,” JAMA 322 (2019): 30-32.

[62] M. T. Brown, F. T. Murphy, D. M. Radin, I. Davignon, M. D. Smith, and C. R. West, “Tanezumab Reduces Osteoarthritic Knee Pain: Results of a Randomized, Double-Blind, Placebo-Controlled Phase III Trial,” Journal of Pain 13 (2012): 790-798.

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2024 The Author(s). Cell Proliferation published by Beijing Institute for Stem Cell and Regenerative Medicine and John Wiley & Sons Ltd.