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Equilibrative Nucleoside Transporter 1 is a Target to Modulate Neuroinflammation and Improve Functional Recovery in Mice with Spinal Cord Injury

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Abstract

Neuroinflammation plays a critical role in the neurological recovery of spinal cord injury (SCI). Adenosine can modulate neuroinflammation, whose uptake is mediated by nucleoside transporters. This study aimed to investigate the roles of equilibrative nucleoside transporter 1 (Ent1) in the inflammatory responses and functional recovery of SCI. Spinal cord contusion at the T10 dorsal portion was induced in mice to cause partial paralysis of the hindlimbs. Genetic deletion and pharmacological inhibition of Ent1 were used to evaluate the role of Ent1 in SCI. The outcomes were evaluated in terms of the Basso Mouse Scale (BMS), gait analysis, astrogliosis, microgliosis, and cytokine levels on day 14 post-injury. As a result, Ent1 deletion reduced neuroinflammation and improved the BMS score (4.88 ± 0.35 in Ent1−/− vs. 3.78 ± 1.09 in Ent1+/+) and stride length (3.74 ± 0.48 cm in Ent1−/− vs. 2.82 ± 0.78 cm in Ent1+/+) of mice with SCI. Along with the reduced lesion size, more preserved neurons were identified in the perilesional area of mice with Ent1 deletion (102 ± 23 in Ent1−/− vs. 73 ± 10 in Ent1+/+). The results of pharmacological inhibition were consistent with the findings of genetic deletion. Moreover, Ent1 inhibition decreased the protein level of complement 3 (an A1 marker), but increased the levels of S100 calcium-binding protein a10 (an A2 marker) and transforming growth factor-β, without changing the levels of inducible nitric oxide synthase (a M1 marker) and arginase 1 (a M2 marker) at the injured site. These findings indicate the important role of Ent1 in the pathogenesis and treatment of SCI.

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Data Availability

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

References

  1. Brown CM, Mulcahey TA, Filipek NC, Wise PM (2010) Production of proinflammatory cytokines and chemokines during neuroinflammation: novel roles for estrogen receptors alpha and beta. Endocrinology 151:4916–4925

    Article  CAS  Google Scholar 

  2. David S, Kroner A (2015) Inflammation and secondary damage after spinal cord injury, in Neural Regeneration (So KF and Xu XM eds) pp245–264, Academic Press

  3. Hausmann ON (2003) Post-traumatic inflammation following spinal cord injury. Spinal Cord 41:369–378

    Article  CAS  Google Scholar 

  4. Anwar MA, Al Shehabi TS, Eid AH (2016) Inflammogenesis of secondary spinal cord injury. Front Cell Neurosci 10:98

    Article  Google Scholar 

  5. Bellver-Landete V, Bretheau F, Mailhot B, Vallières N, Lessard M, Janelle ME, Vernoux N, Tremblay ME et al (2019) Microglia are an essential component of the neuroprotective scar that forms after spinal cord injury. Nat Commun 10:518

    Article  CAS  Google Scholar 

  6. Tang Y, Le W (2016) Differential roles of M1 and M2 microglia in neurodegenerative diseases. Mol Neurobiol 53:1181–1194

    Article  CAS  Google Scholar 

  7. Okada S, Nakamura M, Katoh H, Miyao T, Shimazaki T, Ishii K, Yamane J, Yoshimura A et al (2006) Conditional ablation of Stat3 or Socs3 discloses a dual role for reactive astrocytes after spinal cord injury. Nat Med 12:829–834

    Article  CAS  Google Scholar 

  8. Rolls A, Shechter R, Schwartz M (2009) The bright side of the glial scar in CNS repair. Nat Rev Neurosci 10:235–241

    Article  CAS  Google Scholar 

  9. Liddelow SA, Barres BA (2017) Reactive astrocytes: production, function, and therapeutic potential. Immunity 46:957–967

    Article  CAS  Google Scholar 

  10. Diniz LP, Matias IC, Garcia MN, Gomes FC (2014) Astrocytic control of neural circuit formation: highlights on TGF-beta signaling. Neurochem Int 78:18–27

    Article  CAS  Google Scholar 

  11. Eltzschig HK, Sitkovsky MV, Robson SC (2012) Purinergic signaling during inflammation. N Engl J Med 367:2322–2333

    Article  CAS  Google Scholar 

  12. Seydyousefi M, Moghanlou AE, Metz GAS, Gursoy R, Faghfoori MH, Mirghani SJ, Faghfoori Z (2019) Exogenous adenosine facilitates neuroprotection and functional recovery following cerebral ischemia in rats. Brain Res Bull 153:250–256

    Article  CAS  Google Scholar 

  13. McAdoo DJ, Robak G, Xu GY, Hughes MG (2000) Adenosine release upon spinal cord injury. Brain Res 854:152–157

    Article  CAS  Google Scholar 

  14. Golder FJ, Ranganathan L, Satriotomo I, Hoffman M, Lovett-Barr MR, Watters JJ, Baker-Herman TL, Mitchell GS (2008) Spinal adenosine A2a receptor activation elicits long-lasting phrenic motor facilitation. J Neurosci 28:2033–2042

    Article  CAS  Google Scholar 

  15. Irrera N, Arcoraci V, Mannino F, Vermiglio G, Pallio G, Minutoli L, Bagnato G, Anastasi GP et al (2018) Activation of A2A receptor by PDRN reduces neuronal damage and stimulates WNT/β-CATENIN driven neurogenesis in spinal cord injury. Front Pharmacol 9:506

    Article  Google Scholar 

  16. Xu S, Wang J, Zhong J, Shao M, Jiang J, Song J, Zhu W, Zhang F et al (2021) CD73 alleviates GSDMD-mediated microglia pyroptosis in spinal cord injury through PI3K/AKT/Foxo1 signaling. Clin Transl Med 11:e269

    Article  CAS  Google Scholar 

  17. Ciruela F, Ferré S, Casadó V, Cortés A, Cunha RA, Lluis C, Franco R (2006) Heterodimeric adenosine receptors: a device to regulate neurotransmitter release. Cell Mol Life Sci 63:2427–2431

    Article  CAS  Google Scholar 

  18. Pastor-Anglada M, Pérez-Torras S (2018) Emerging roles of nucleoside transporters. Front Pharmacol 9:606

    Article  Google Scholar 

  19. Ackley MA, Governo RJ, Cass CE, Young JD, Baldwin SA, King AE (2003) Control of glutamatergic neurotransmission in the rat spinal dorsal horn by the nucleoside transporter ENT1. J Physiol 548:507–517

    Article  CAS  Google Scholar 

  20. Sunkin SM, Ng L, Lau C, Dolbeare T, Gilbert TL, Thompson CL, Hawrylycz M, Dang C (2013) Allen Brain Atlas: an integrated spatio-temporal portal for exploring the central nervous system. Nucleic Acids Res 41:D996–D1008

    Article  CAS  Google Scholar 

  21. Kao YH, Lin MS, Chen CM, Wu YR, Chen HM, Lai HL, Chern Y, Lin CJ (2017) Targeting ENT1 and adenosine tone for the treatment of Huntington’s disease. Hum Mol Genet 26:467–478

    CAS  Google Scholar 

  22. Chang CP, Chang YG, Chuang PY, Nguyen TNA, Wu KC, Chou FY, Cheng SJ, Chen HM et al (2021) Equilibrative nucleoside transporter 1 inhibition rescues energy dysfunction and pathology in a model of tauopathy. Acta Neuropathol Commun 9:112

    Article  CAS  Google Scholar 

  23. Basso DM, Fisher LC, Anderson AJ, Jakeman LB, McTigue DM, Popovich PG (2006) Basso Mouse Scale for locomotion detects differences in recovery after spinal cord injury in five common mouse strains. J Neurotrauma 23:635–659

    Article  Google Scholar 

  24. Lai PH, Wang TH, Zhang NY, Wu KC, Yao CJ, Lin CJ (2021) Changes of blood-brain-barrier function and transfer of amyloid beta in rats with collagen-induced arthritis. J Neuroinflammation 18:35

    Article  CAS  Google Scholar 

  25. Porter AG, Jänicke RU (1999) Emerging roles of caspase-3 in apoptosis. Cell Death Differ 6:99–104

    Article  CAS  Google Scholar 

  26. Boghdadi AG, Teo L, Bourne JA (2020) The neuroprotective role of reactive astrocytes after central nervous system injury. J Neurotrauma 37:681–691

    Article  Google Scholar 

  27. Yi JJ, Barnes AP, Hand R, Polleux F, Ehlers MD (2010) TGF-beta signaling specifies axons during brain development. Cell 142:144–157

    Article  CAS  Google Scholar 

  28. Satake K, Matsuyama Y, Kamiya M, Kawakami H, Iwata H, Adachi K, Kiuchi K (2000) Nitric oxide via macrophage iNOS induces apoptosis following traumatic spinal cord injury. Brain Res Mol Brain Res 85:114–122

    Article  CAS  Google Scholar 

  29. Orr MB, Gensel JC (2018) Spinal cord injury scarring and inflammation: therapies targeting glial and inflammatory responses. Neurotherapeutics 15:541–553

    Article  CAS  Google Scholar 

  30. Hurlbert RJ, Hadley MN, Walters BC, Aarabi B, Dhall SS, Gelb DE, Rozzelle CJ, Ryken TC et al (2013) Pharmacological therapy for acute spinal cord injury. Neurosurgery 72(Suppl 2):93–105

    Article  Google Scholar 

  31. Farr SA, Cuzzocrea S, Esposite E, Campolo M, Niehoff M, Doyle TM, Salvemini D (2020) Adenosine A3 receptor as a novel therapeutic target to reduce secondary events and improve neurocognitive functions following traumatic brain injury. J Neuroinflammation 17:339

    Article  CAS  Google Scholar 

  32. Little JW, Ford A, Symons-Liguori AM, Chen Z, Janes K, Doyle T, Xie J, Luongo L et al (2015) Endogenous adenosine A3 receptor activation selectively alleviates persistent pain states. Brain 138:28–35

    Article  Google Scholar 

  33. Guitart X, Bonaventura J, Rea W, Orrú M, Cellai L, Dettori I, Pedata F, Brugarolas M et al (2016) Equilibrative nucleoside transporter ENT1 as a biomarker of Huntington disease. Neurobiol Dis 96:47–53

    Article  CAS  Google Scholar 

  34. Sari Y, Sreemantula SN, Lee MR, Choi DS (2013) Ceftriaxone treatment affects the levels of GLT1 and ENT1 as well as ethanol intake in alcohol-preferring rats. J Mol Neurosci 51:779–787

    Article  CAS  Google Scholar 

  35. Lee CC, Chang CP, Lin CJ, Lai HL, Kao YH, Cheng SJ, Chen HM, Liao YP et al (2018) Adenosine augmentation evoked by an ENT1 inhibitor improves memory impairment and neuronal plasticity in the APP/PS1 mouse model of Alzheimer’s disease. Mol Neurobiol 55:8936–8952

    Article  CAS  Google Scholar 

  36. Keil GJ, DeLander GE (1995) Time-dependent antinociceptive interactions between opioids and nucleoside transport inhibitors. J Pharmacol Exp Ther 274:1387–1392

    CAS  Google Scholar 

  37. Han D, Yu Z, Liu W, Yin D, Pu Y, Feng J, Yuan Y, Huang A et al (2018) Plasma Hemopexin ameliorates murine spinal cord injury by switching microglia from the M1 state to the M2 state. Cell Death Dis 9:1–16

    Article  Google Scholar 

  38. Kwak EK, Kim JW, Kang KS, Lee YH, Hua QH, Park TI, Park JY, Sohn YK (2005) The role of inducible nitric oxide synthase following spinal cord injury in rat. J Korean Med Sci 20:663–669

    Article  CAS  Google Scholar 

  39. Hinton DJ, Lee MR, Jang JS, Choi DS (2014) Type 1 equilibrative nucleoside transporter regulates astrocyte-specific glial fibrillary acidic protein expression in the striatum. Brain Behav 4:903–914

    Article  Google Scholar 

  40. Vismara I, Papa S, Veneruso V, Mauri E, Mariani A, De Paola M, Affatato R, Rossetti A et al (2020) Selective modulation of A1 astrocytes by drug-loaded nano-structured gel in spinal cord injury. ACS Nano 14:360–371

    Article  CAS  Google Scholar 

  41. Wang L, Pei S, Han L, Guo B, Li Y, Duan R, Yao Y, Xue B et al (2018) Mesenchymal stem cell-derived exosomes reduce A1 astrocytes via downregulation of phosphorylated NFκB P65 subunit in spinal cord injury. Cell Physiol Biochem 50:1535–1559

    Article  CAS  Google Scholar 

  42. Neal M, Luo J, Harischandra DS, Gordon R, Sarkar S, Jin H, Anantharam V, Désaubry L et al (2018) Prokineticin-2 promotes chemotaxis and alternative A2 reactivity of astrocytes. Glia 66:137–2157

    Article  Google Scholar 

  43. Abdipranoto-Cowley A, Park JS, Croucher D, Daniel J, Henshall S, Galbraith S, Mervin K, Vissel B (2009) Activin A is essential for neurogenesis following neurodegeneration. Stem Cells 27:1330–1346

    Article  CAS  Google Scholar 

Download references

Funding

This work was supported in part by Grant no. NTPC110-002 from the New Taipei City Hospital of Taiwan and by Grant no. AS-KPQ-111-KNT from the Academia Sinica.

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Author notes

  1. Kuan-Yu Chen and Chiao-Shin Lu contributed equally to this work.

Authors and Affiliations

  1. New Taipei City Hospital, Taipei, Taiwan

    Kuan-Yu Chen

  2. Institute of Medical Sciences, Tzu Chi University, Hualien, Taiwan

    Kuan-Yu Chen & Cheng-Yoong Pang

  3. School of Pharmacy, College of Medicine, National Taiwan University, Taipei, Taiwan

    Chiao-Shin Lu, Chin-Jui Ho, Kuo-Chen Wu & Chun-Jung Lin

  4. Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan

    Kuo-Chen Wu, Hsin-Lin Lai & Yijuang Chern

  5. Department of Chemical Engineering, College of Engineering, National Taiwan University, Taipei, Taiwan

    Hsiu-Wei Yang

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Contributions

K-Y C contributed to the conception and design and funding acquisition; C-S L contributed to the conception and design, acquisition and analysis of data, and manuscript preparation; C-Y P contributed to the conception and design and advised; C-J H, K-C W, H-W Y, and H–L L contributed to the acquisition and analysis of data; Y C contributed to the conception and design and advised; C-J L contributed to the conception and design, advised, manuscript preparation, and funding acquisition. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Chun-Jung Lin.

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All the animal experiments were approved by the Institutional Animal Care and Use Committee of the National Taiwan University College of Medicine (IACUC number: 20200073) and followed the 3Rs principle.

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Chen, KY., Lu, CS., Pang, CY. et al. Equilibrative Nucleoside Transporter 1 is a Target to Modulate Neuroinflammation and Improve Functional Recovery in Mice with Spinal Cord Injury. Mol Neurobiol 60, 369–381 (2023). https://doi.org/10.1007/s12035-022-03080-2

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