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Effects of FTY720 on Neural Cell Behavior in Two and Three-Dimensional Culture and in Compression Spinal Cord Injury

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Abstract

Introduction

The present study aimed to evaluate the effects of FTY720 as a neuromodulatory drug on the behaviors of neural stem/progenitor cells (NS/PCs) in two-dimensional (2-D) and three-dimensional (3-D) cultures and in spinal cord injury (SCI).

Methods

The NS/PCs isolated from the ganglionic eminence of the 13.5-day old embryos were cultured as free-floating spheres. The single cells obtained from the second passage were cultured in 96-well plates without any scaffold (2-D) or containing PuraMatrix (PM, 3-D) or were used for transplantation in a mouse model of compression SCI. After exposure to 0, 10, 50, and 100 nanomolar of FTY720, the survival, proliferation, and migration of the NS/PCs were evaluated in vitro using MTT assay, neurosphere assay, and migration assay, respectively. Moreover, the functional recovery, survival and migration capacity of transplanted cells exposure to 100 nanomolar FTY720 were investigated in SCI.

Results

Cell survival and migration capacity increased after exposure to 50 and 100 nanomolar FTY720. In addition, higher doses of FTY720 led to the formation of more extensive and more neurospheres. Although this phenomenon was similar in both 2-D and 3-D cultures, PM induced better distribution of the cells in a 3-D environment. Furthermore, co-administration of FTY720 and NS/PCs 7 days after SCI enhanced functional recovery and both survival and migration of transplanted cells in the lesion site.

Conclusions

Due to the positive effects of FTY720 on the behavior of NS/PCs, using them in combination therapies can be an appealing approach for stem cell therapy in CNS injury.

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References

  1. Ahuja, C. S., J. R. Wilson, S. Nori, M. R. Kotter, C. Druschel, A. Curt, and M. G. Fehlings. Traumatic spinal cord injury. Nat. Rev. Dis. Primers. 3:1–21, 2017.

    Article  Google Scholar 

  2. Aligholi, H., S. M. Rezayat, H. Azari, S. E. Mehr, M. Akbari, S. M. M. Mousavi, et al. Preparing neural stem/progenitor cells in PuraMatrix hydrogel for transplantation after brain injury in rats. A comparative methodological study. Brain Res. 1642:197–208, 2016.

    Article  Google Scholar 

  3. Azari, H., S. Sharififar, M. Rahman, S. Ansari, and B. A. Reynolds. Establishing embryonic mouse neural stem cell culture using the neurosphere assay. J. Vis. Exp. 47:2457, 2011.

    Google Scholar 

  4. Basso, D. M., L. C. Fisher, A. J. Anderson, L. B. Jakeman, D. M. Mctigue, and P. G. Popovich. Basso Mouse Scale for locomotion detects differences in recovery after spinal cord injury in five common mouse strains. J. Neurotrauma. 23(5):635–659, 2006.

    Article  Google Scholar 

  5. Blaho, V. A., and T. Hla. An update on the biology of sphingosine 1-phosphate receptors. J. Lipid Res. 55(8):1596–1608, 2014.

    Article  Google Scholar 

  6. Brait, V. H., G. Tarrasón, A. Gavaldà, N. Godessart, and A. M. Planas. Selective sphingosine 1-phosphate receptor 1 agonist is protective against ischemia/reperfusion in mice. Stroke. 47(12):3053–3056, 2016.

    Article  Google Scholar 

  7. Choo, A. M., J. Liu, M. Dvorak, W. Tetzlaff, and T. R. Oxland. Secondary pathology following contusion, dislocation, and distraction spinal cord injuries. Exp. Neurol. 212(2):490–506, 2008.

    Article  Google Scholar 

  8. Danielyan, L., M. Schwab, G. Siegel, B. Brawek, O. Garaschuk, and N. Asavapanumas. Cell motility and migration as determinants of stem cell efficacy. EBioMedicine. 60:102989, 2020.

    Article  Google Scholar 

  9. Gage, F. H. Mammalian neural stem cells. Science. 287(5457):1433–1438, 2000.

    Article  Google Scholar 

  10. Harada, J., M. Foley, M. A. Moskowitz, and C. Waeber. Sphingosine-1-phosphate induces proliferation and morphological changes of neural progenitor cells. J. Neurochem. 88(4):1026–1039, 2004.

    Article  Google Scholar 

  11. Hasegawa, Y., H. Suzuki, T. Sozen, W. Rolland, and J. H. Zhang. Activation of sphingosine 1-phosphate receptor-1 by FTY720 is neuroprotective after ischemic stroke in rats. Stroke. 41(2):368–374, 2010.

    Article  Google Scholar 

  12. Hemmati, F., L. Dargahi, S. Nasoohi, R. Omidbakhsh, Z. Mohamed, Z. Chik, et al. Neurorestorative effect of FTY720 in a rat model of Alzheimer’s disease: comparison with memantine. Behav. Brain Res. 252:415–421, 2013.

    Article  Google Scholar 

  13. Kappos, L., J. Antel, G. Comi, X. Montalban, P. O’connor, and C. H. Polman. Oral fingolimod (FTY720) for relapsing multiple sclerosis. N. Engl. J. Med. 355(11):1124–1140, 2006.

    Article  Google Scholar 

  14. Kassmer, S. H., D. Rodriguez, A. D. Langenbacher, C. Bui, and A. W. De Tomaso. Migration of germline progenitor cells is directed by sphingosine-1-phosphate signalling in a basal chordate. Nat. Commun. 6(1):1–10, 2015.

    Article  Google Scholar 

  15. Kim, S. U., H. J. Lee, and Y. B. Kim. Neural stem cell-based treatment for neurodegenerative diseases. Neuropathology. 33(5):491–504, 2013.

    Google Scholar 

  16. Kimura, A., T. Ohmori, R. Ohkawa, S. Madoiwa, J. Mimuro, T. Murakami, et al. Essential roles of sphingosine 1-phosphate/S1P1 receptor axis in the migration of neural stem cells toward a site of spinal cord injury. Stem Cells. 25(1):115–124, 2007.

    Article  Google Scholar 

  17. Kong, W., Z. Qi, P. Xia, Y. Chang, H. Li, Y. Qu, et al. Local delivery of FTY720 and NSCs on electrospun PLGA scaffolds improves functional recovery after spinal cord injury. RSC Adv. 9(31):17801–17811, 2019.

    Article  Google Scholar 

  18. Lee, K. D., W. N. Chow, C. Sato-Bigbee, M. R. Graf, R. S. Graham, R. J. Colello, et al. FTY720 reduces inflammation and promotes functional recovery after spinal cord injury. J. Neurotrauma. 26(12):2335–2344, 2009.

    Article  Google Scholar 

  19. Lu, L., A. H. Barfejani, T. Qin, Q. Dong, C. Ayata, and C. Waeber. Fingolimod exerts neuroprotective effects in a mouse model of intracerebral hemorrhage. Brain Res. 1555:89–96, 2014.

    Article  Google Scholar 

  20. Marsh, S. E., and M. Blurton-Jones. Neural stem cell therapy for neurodegenerative disorders: the role of neurotrophic support. Neurochem. Int. 106:94–100, 2017.

    Article  Google Scholar 

  21. Negah, S. S., Z. Khaksar, H. Aligholi, S. M. Sadeghi, S. M. M. Mousavi, H. Kazemi, et al. Enhancement of neural stem cell survival, proliferation, migration, and differentiation in a novel self-assembly peptide nanofibber scaffold. Mol. Neurobiol. 54(10):8050–8062, 2017.

    Article  Google Scholar 

  22. Noda, H., H. Takeuchi, T. Mizuno, and A. Suzumura. Fingolimod phosphate promotes the neuroprotective effects of microglia. J. Neuroimmunol. 256(1–2):13–18, 2013.

    Article  Google Scholar 

  23. Norimatsu, Y., T. Ohmori, A. Kimura, S. Madoiwa, J. Mimuro, A. Seichi, et al. FTY720 improves functional recovery after spinal cord injury by primarily nonimmunomodulatory mechanisms. Am. J. Pathol. 180(4):1625–1635, 2012.

    Article  Google Scholar 

  24. Pébay, A., M. Toutant, J. Prémont, C. F. Calvo, L. Venance, J. Cordier, et al. Sphingosine-1-phosphate induces proliferation of astrocytes: regulation by intracellular signalling cascades. Eur. J. Neurosci. 13(11):2067–2076, 2001.

    Article  Google Scholar 

  25. Pham, T.-C.T., J. I. Fells Sr., D. A. Osborne, E. J. North, M. M. Naor, and A. L. Parrill. Molecular recognition in the sphingosine 1-phosphate receptor family. J. Mol. Graphics Model. 26(8):1189–1201, 2008.

    Article  Google Scholar 

  26. Pinschewer, D. D., A. F. Ochsenbein, B. Odermatt, V. Brinkmann, H. Hengartner, and R. M. Zinkernagel. FTY720 immunosuppression impairs effector T cell peripheral homing without affecting induction, expansion, and memory. J. Immunol. 164(11):5761–5770, 2000.

    Article  Google Scholar 

  27. Shamsi, F., Z. Zeraatpisheh, H. Alipour, A. Nazari, and H. Aligholi. The effects of minocycline on proliferation, differentiation and migration of neural stem/progenitor cells. Int. J. Neurosci. 130(6):601–609, 2020.

    Article  Google Scholar 

  28. Singh, I. N., and E. D. Hall. Multifaceted roles of sphingosine-1-phosphate: How does this bioactive sphingolipid fit with acute neurological injury? J. Neurosci. Res. 86(7):1419–1433, 2008.

    Article  Google Scholar 

  29. Tan, B., Z. Luo, Y. Yue, Y. Liu, L. Pan, L. Yu, and Y. Yin. Effects of FTY720 (fingolimod) on proliferation, differentiation, and migration of brain-derived neural stem cells. Stem Cells Int. 2016.

  30. Wang, J., J. Wang, P. Lu, Y. Cai, Y. Wang, L. Hong, et al. Local delivery of FTY720 in PCL membrane improves SCI functional recovery by reducing reactive astrogliosis. Biomaterials. 62:76–87, 2015.

    Article  Google Scholar 

  31. Wei, Y., M. Yemisci, H. H. Kim, L. M. Yung, H. K. Shin, S. K. Hwang, et al. Fingolimod provides long-term protection in rodent models of cerebral ischemia. Ann. Neurol. 69(1):119–129, 2011.

    Article  Google Scholar 

  32. Willis, M. A., and J. A. Cohen. Fingolimod therapy for multiple sclerosis. Paper presented at the Seminars in Neurology, 2013.

  33. Zeraatpisheh, Z., E. Mirzaei, M. Nami, H. Alipour, S. Ghasemian, and H. Azari et al. A new and simple method for spinal cord injury induction in mice. Basic Clin. Neurosci. 2020, forthcoming.

  34. Zeraatpisheh, Z., E. Mirzaei, M. Nami, H. Alipour, M. Mahdavipour, P. Sarkoohi, et al. Local delivery of Fingolimod through PLGA nanoparticles and PuraMatrix-embedded neural precursor cells promote motor function recovery and tissue repair in spinal cord injury. Eur. J. Neurosci. 54:5620, 2021.

    Article  Google Scholar 

  35. Zhang, S., X. Zhao, and L. Spirio. PuraMatrix: self-assembling peptide nanofiber scaffolds. Scaffolding Tissue Eng. 2005, 217–238.

  36. Zhang, Y., X. Li, B. Ciric, C.-G. Ma, B. Gran, A. Rostami, and G.-X. Zhang. Effect of fingolimod on neural stem cells: a novel mechanism and broadened application for neural repair. Mol. Ther. 25(2):401–415, 2017.

    Article  Google Scholar 

Download references

Acknowledgments

The authors appreciate the support from Shiraz University of Medical Sciences, Shiraz, Iran.

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical Approval

All animal care and experimental procedures were conducted according to the National Institute of Health guidelines on animal care and use and were approved by the Institutional Ethics Committee of Shiraz University of Medical Sciences (approval no. 11659).

Research Involving Human Rights

The article does not contain any studies with human participants by any of the authors.

Author information

Authors and Affiliations

  1. Department of Neuroscience, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran

    Zahra Zeraatpisheh, Fatemeh Shamsi & Hadi Aligholi

  2. Epilepsy Research Center, Shiraz University of Medical Sciences, Shiraz, Iran

    Zahra Zeraatpisheh

  3. Department of Pharmacology and Toxicology, School of Pharmacy, Hormozgan University of Medical Sciences, Bandar Abbas, Iran

    Parisa Sarkoohi

  4. Department of Anatomical Sciences, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

    Somayyeh Torabi

  5. Department of Tissue Engineering & Applied cell Sciences, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran

    Hamed Alipour

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  1. Zahra Zeraatpisheh
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Correspondence to Hadi Aligholi.

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Associate Editor Stephanie Michelle Willerth oversaw the review of this article.

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Zeraatpisheh, Z., Shamsi, F., Sarkoohi, P. et al. Effects of FTY720 on Neural Cell Behavior in Two and Three-Dimensional Culture and in Compression Spinal Cord Injury. Cel. Mol. Bioeng. 15, 331–340 (2022). https://doi.org/10.1007/s12195-022-00724-0

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  • DOI: https://doi.org/10.1007/s12195-022-00724-0

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