iTRAQ-based proteomics profiling of Schwann cells before and after peripheral nerve injury

Document Type : Original Article

Authors

1 Department of Orthopedics, Tianjin Medical University General Hospital, 154 Anshan Road, Heping District, Tianjin, China

2 Tianjin Neurological Institute, Key Laboratory of Post-Neuroinjury Neuro-repair and Regeneration in Central Nervous System, Ministry of Education and Tianjin City, Tianjin, China

Abstract

Objective(s): Schwann cells (SCs) have a wide range of applications as seed cells in the treatment of nerve injury during transplantation. However, there has been no report yet on kinds of proteomics changes that occur in Schwann cells before and after peripheral nerve injury.
Materials and Methods: Activated Schwann cells (ASCs) and normal Schwann cells (NSCs) were obtained from adult Wistar rat sciatic nerves. After immunofluorescence identification, we identified differentially expressed proteins in the ASCs and NSCs using isobaric tags for relative and absolute quantitation (iTRAQ) combined with high-resolution Orbitrap liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). In addition, all the differentially expressed proteins were analyzed by Gene ontology (GO) analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis. Finally, several differentially expressed proteins were selected for Western blot verification.
Results: A total of 122 differentially expressed proteins in ASCs and NSCs were screened. GO analysis suggested that these different proteins are likely to accumulate in the cytoplasm and are associated with single-multicellular organism processes. The KEGG pathway analysis suggested that proteins related to purine metabolism were significantly enriched. The expression of Transmembrane glycoprotein NMB (GPNMB), Ectonucleotide pyrophosphatase/phosphodiesterase family member 3 (ENPP3), and other proteins were consistent with the proteomics data obtained by Western blot analysis.
Conclusion: GPNMB, ENPP3, GFPT2, and other proteins may play an important role in the repair of peripheral nerve injury. This study may provide new insights into changes in SCs after peripheral nerve injury.

Keywords

Main Subjects

References

1. Birbeck GL, Meyer AC, Ogunniyi A. Nervous system disorders across the life course in resource-limited settings. Nature 2015; 527:167-171.
2. Martinez AR, Faber I, Nucci A, Appenzeller S, Franca MC. Autoimmune neuropathies associated to rheumatic diseases. Autoimmun Rev 2017; 16:335-342.
3. Huang H, Mao G, Chen L, Liu A. Progress and challenges with clinical cell therapy in neurorestoratology. J Neurorestoratol 2015; 3:91-95.
4. Gross G, Eshhar Z. Therapeutic potential of T-cell chimeric antigen receptors (CARs) in cancer treatment: counteracting off-tumor toxicities for safe CAR T-cell therapy. Annu Rev Pharmacol Toxicol 2016; 56:59-83.
5. Sarkar P, Rice CM, Scolding NJ. Cell therapy for multiple sclerosis. CNS Drugs 2017; 31:453-469.
6. Ansari S, Seagroves JT, Chen C, Shah K, Aghaloo T, Wu BM, et al. Dental and orofacial mesenchymal stem cells in craniofacial regeneration: the prosthodontist’s point of view. J Prosthet Dent 2017; 118:455-461.
7. Boilly B, Faulkner S, Jobling P, Hondermarck H. Nerve dependence: from regeneration to cancer. Cancer Cell 2007; 31:342-354.
8. Zawadzka M, Rivers LE, Fancy SP, Zhao C, Tripathi R, Jamen F, et al. CNS-resident glial progenitor/stem cells produce Schwann cells as well as oligodendrocytes during repair of CNS demyelination. Cell Stem Cell 2010; 6:578-590.
9. Koenig HL, Schumacher M, Ferzaz B, Thi AN, Ressouches A, Guennoun R, et al. Progesterone synthesis and myelin formation by Schwann cells. Science 1995; 268:1500-1503.
10. Pearse DD, Pereira FC, Marcillo AE, Bates ML, Berrocal YA, Filbin MT, et al. cAMP and Schwann cells promote axonal growth and functional recovery after spinal cord injury. Nat Med 2004; 10:610-616.
11. Park HW, Lim MJ, Jung H, Lee SP, Paik KS, Chang MS. Human mesenchymal stem cell‐derived Schwann cell‐like cells exhibit neurotrophic effects, via distinct growth factor production, in a model of spinal cord injury. Glia 2010; 58:1118-1132.
12. Carlson KB, Singh P, Feaster MM, Ramnarain A, Pavlides C, Chen ZL, et al. Mesenchymal stem cells facilitate axon sorting, myelination, and functional recovery in paralyzed mice deficient in Schwann cell‐derived laminin. Glia 2011; 59:267-277.
13. Lai BQ, Che MT, Du BL, Zeng X, Ma YH, Feng B, et al. Transplantation of tissue engineering neural network and formation of neuronal relay into the transected rat spinal cord. Biomaterials 2016; 109:40-54.
14. Lavdas AA, Papastefanaki F, Thomaidou D, Matsas R. Cell adhesion molecules in gene and cell therapy approaches for nervous system repair. Curr Gene Ther 2011; 11:90-100.
15. Yi S, Tang X, Yu J, Liu J, Ding F, Gu X. Microarray and qPCR analyses of wallerian degeneration in rat sciatic nerves. Front Cell Neurosci 2017; 11:22.
16. Yu J, Gu X, Yi S. Ingenuity pathway analysis of gene expression profiles in distal nerve stump following nerve injury: insights into wallerian degeneration. Front Cell Neurosci 2016; 10:274.
17. Zhang L, Jia X, Jin JO, Lu H, Tan Z. Recent 5-year findings and technological advances in the proteomic study of HIV-associated disorders. Genomics Proteomics Bioinformatics 2017; 15:110-120.
18. Zhang P, Zhu S, Li Y, Zhao M, Liu M, Gao J, et al. Quantitative proteomics analysis to identify diffuse axonal injury biomarkers in rats using iTRAQ coupled LC–MS/MS. J Proteom 2016; 133:93-99.
19. Chen J, Ge L, Liu A, Yuan Y, Ye J, Zhong J, et al. Identification of pathways related to FAF1/H. pylori-associated gastric carcinogenesis through an integrated approach based on iTRAQ quantification and literature review. J Proteom 2016; 131:163-176.
20. Mehrotra S, Goyal V. Evaluation of designer crops for biosafety—A scientist’s perspective. Gene 2013; 515:241-248.
21. Yang S, Pei Y, Zhao A. iTRAQ-based proteomic analysis of porcine kidney epithelial PK15 cells infected with pseudorabies virus. Sci Rep 2017; 7:45922.
22. Woodhoo A, Alonso MBD, Droggiti A, Turmaine M, D’antonio M, Parkinson DB, et al. Notch controls embryonic Schwann cell differentiation, postnatal myelination and adult plasticity. Nat Neurosci 2009; 12:839-847.
23. Keilhoff G, Fansa H, Schneider W, Wolf G. In vivo predegeneration of peripheral nerves: an effective technique to obtain activated Schwann cells for nerve conduits. J Neurosci Methods 1999; 89:17-24.
24. Zhou XH, Lin W, Ren YM, Liu S, Fan BY, Wei ZJ, et al. Comparison of DNA methylation in Schwann cells before and after peripheral nerve injury in rats. BioMed Res Int 2017; 2017.
25. Hu X, Li N, Wu L, Li C, Li C, Zhang L, et al. Quantitative iTRAQ-based proteomic analysis of phosphoproteins and ABA-regulated phosphoproteins in maize leaves under osmotic stress. Sci Rep 2015; 5:15626.
26. Ma J, Yao Y, Wang P, Liu Y, Zhao L, Li Z, et al. MiR-152 functions as a tumor suppressor in glioblastoma stem cells by targeting Kruppel-like factor 4. Cancer Lett 2014; 355:85-95.
27. Burnside MN, Pyatt RE, Hughes A, Baker PB, Pierson CR. Complex brain malformations associated with chromosome 6q27 gain that includes THBS2, which encodes thrombospondin 2, an astrocyte-derived protein of the extracellular matrix. Pediatr Dev Pathol 2015; 18:59-65.
28. Akane H, Saito F, Shiraki A, Imatanaka N, Akahori Y, Itahashi M, et al. Gene expression profile of brain regions reflecting aberrations in nervous system development targeting the process of neurite extension of rat offspring exposed developmentally to glycidol. J Appl Toxicol 2014; 34:1389-1399.
29. Huber RJ. Using the social amoeba Dictyostelium to study the functions of proteins linked to neuronal ceroid lipofuscinosis. Int J Biomed Sci 2016; 23:83.
30. Nonoda Y, Saito Y, Nagai S, Sasaki M, Iwasaki T, Matsumoto N, et al. Progressive diffuse brain atrophy in West syndrome with marked hypomyelination due to SPTAN1 gene mutation. Brain Dev 2013; 35:280-283.
31. Cáceres M, Suwyn C, Maddox M, Thomas JW, Preuss TM. Increased cortical expression of two synaptogenic thrombospondins in human brain evolution. Cereb Cortex 2006; 17:2312-2321.
32. Pasquini LA, Millet V, Hoyos HC, Giannoni JP, Croci DO, Marder M, et al. Galectin-3 drives oligodendrocyte differentiation to control myelin integrity and function. Cell Death Differ 2011; 18:1746-1756.
33. Mutka AL, Haapanen A, Käkelä R, Lindfors M, Wright AK, Inkinen T, et al. Murine cathepsin D deficiency is associated with dysmyelination/myelin disruption and accumulation of cholesteryl esters in the brain. J Neurochem 2010; 112:193-203.
34. Bollen M, Gijsbers R, Ceulemans H, Stalmans W, Stefan C. Nucleotide pyrophosphatases/phosphodiesterases on the move. Crit Rev Biochem Mol Biol 2000; 35:393-432.
35. Jankowski M, Piwkowska A, Rogacka D, Audzeyenka I, Janaszak-Jasiecka A, Angielski S. Expression of membrane-bound NPP-type ecto-phosphodiesterases in rat podocytes cultured at normal and high glucose concentrations. Biochem biophys res commun 2011; 416:64-69.
36. Vuaden FC, Savio LE, Ramos DB, Casali EA, Bogo MR, Bonan CD. Endotoxin-induced effects on nucleotide catabolism in mouse kidney. Eur J Pharmacol 2012; 674:422-429.
37. Gutknecht M, Geiger J, Joas S, Dorfel D, Salih HR, Muller MR, et al. The transcription factor MITF is a critical regulator of GPNMB expression in dendritic cells. Cell commun signal 2015; 13:19.
38. Weiss T, Taschner-Mandl S, Bileck A, Slany A, Kromp F, Rifatbegovic F, et al. Proteomics and transcriptomics of peripheral nerve tissue and cells unravel new aspects of the human Schwann cell repair phenotype. Glia 2016; 64:2133-2153.
39. Shen M, Ji Y, Zhang S, Shi H, Chen G, Gu X, et al. A proteome map of primary cultured rat Schwann cells. J Proteome Sci 2012; 10:20.
40. Aghamaleky Sarvestany A, Hunter G, Tavendale A, Lamont DJ, Llavero Hurtado M, Graham LC, et al. Label-free quantitative proteomic profiling identifies disruption of ubiquitin homeostasis as a key driver of Schwann cell defects in spinal muscular atrophy. J Proteome Res 2014; 13:4546-4557.
41. Kawahara K, Hirata H, Ohbuchi K, Nishi K, Maeda A, Kuniyasu A, et al. The novel monoclonal antibody 9F5 reveals expression of a fragment of GPNMB/osteoactivin processed by furin-like protease(s) in a subpopulation of microglia in neonatal rat brain. Glia 2016; 64:1938-1961.
42. Coughlin L, Morrison RS, Horner PJ, Inman DM. Mitochondrial morphology differences and mitophagy deficit in murine glaucomatous optic nerve. Invest Ophthalmol Vis Sci 2015; 56:1437-1446.
43. Du J, Liu J, Yao S, Mao H, Peng J, Sun X, et al. Prompt peripheral nerve regeneration induced by a hierarchically aligned fibrin nanofiber hydrogel. Acta biomater 2017; 55:296-309.
44. Liao CP, Pradhan S, Chen Z, Patel AJ, Booker RC, Le LQ. The role of nerve microenvironment for neurofibroma development. Oncotarget 2016; 7:61500-61508.
45. Dey I, Midha N, Singh G, Forsyth A, Walsh SK, Singh B, et al. Diabetic Schwann cells suffer from nerve growth factor and neurotrophin-3 underproduction and poor associability with axons. Glia 2013; 61:1990-1999.
46. Civi S, Emmez G, Dere UA, Borcek AO, Emmez H. Effects of quercetin on chronic constriction nerve injury in an experimental rat model. Acta neurochir 2016; 158:959-965.
47. Wu Z, Ding N, Yu M, Wang K, Luo S, Zou W, et al. Identification of potential biomarkers for rhegmatogenous retinal detachment associated with choroidal detachment by vitreous iTRAQ-based proteomic profiling. Int J Mol Sci 2016; 17:2052.
48. Subbannayya Y, Mir SA, Renuse S, Manda SS, Pinto SM, Puttamallesh VN, et al. Identification of differentially expressed serum proteins in gastric adenocarcinoma. J Proteom 2015; 127:80-88.
49. Wang Y, Liu H, Liang D, Huang Y, Zeng Y, Xing X, et al. Reveal the molecular signatures of hepatocellular carcinoma with different sizes by iTRAQ based quantitative proteomics. J Proteom 2017; 150:230-241.

Iranian Journal of Basic Medical Sciences
Volume 21, Issue 8
August 2018
Pages 832-841

Files

Share

How to cite

Statistics