Human wild-type superoxide dismutase 1 gene delivery to rat bone marrow stromal cells: its importance and potential future trends

Document Type: Original Article

Authors

1 Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

2 Department of Clinical Biochemistry, Faculty of Paramedicine, Ilam University of Medical Sciences, Ilam, Iran

3 Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

4 Shefa Neuroscience Research Center, Khatam-Alanbia Hospital, Tehran, Iran

Abstract

Objective(s): Human superoxide dismutase 1 (SOD1) is the cytosolic form of this enzyme it detoxifies superoxide anions and attenuates their toxicities and concomitant detrimental effects on the cells. It is believed that the amount of these enzymes present in the oxidative stress-induced diseases is crucial for preventing disease progression. Transfection of rat bone marrow stromal cells (BMSCs) by a constructed vector carrying the human wild-type SOD1 gene, a non-viral gene transfer method, was the main aim of this study.
Materials and Methods: For this purpose, the rat BMSCs were transfected with the vector using Turbofect reagent and then stabilized. Western-blot and real-time PCR were also used for evaluation of SOD1 expression.
Results: Data analysis from RT-PCR and Western-blot techniques revealed that the stable transfected cells could secrete human wild-type SOD1 in the supernatant. Also, the total activity of SOD1 was about 0.5±0.09 U/ml and 0.005±0.002 U/ml in the supernatants of the transfected and not-transfected of rat BMSCs, respectively.
Conclusion: This study showed that expansion of the stable transfected rat BMSCs by a constructed vector carrying the human wild-type SOD1 gene is capable of secreting the active SOD1 enzyme under ex-vivo conditions.  The recommendation of this study is that the same experiment would be applicable for expression of the other form of this enzyme, SOD3, as well. More valuable information could probably be provided about the variety of the diseases caused by superoxide anions toxicities by intervention and application of the non-viral method for expressions of SOD1 and SOD3 enzymes.

Keywords

Main Subjects


1. Fontana L, Partridge L, Longo VD. Extending healthy life span--from yeast to humans. Science 2010; 328:321-326
2. Beckman KB,  Ames BN. The free radical theory of aging matures. Physiol Rev 1998; 78:547-581
3. Boveris A, Oshino N, Chance B. The cellular production of hydrogen peroxide. Biochem J 1972; 128: 617-630
4. Alexeyev MF. Is there more to aging than mitochondrial DNA and reactive oxygen species?. Febs J 2009; 276:5768-5787
5. Dowling DK, Simmons LW. Reactive oxygen species as universal constraints in life-history evolution. Proc Biol Sci 2009; 276:1737-1745
6. Speakman JR, Selman C. The free-radical damage theory: Accumulating evidence against a simple link of oxidative stress to ageing and lifespan. Bioessays 2011; 33:255-259
7. Lefer DJ,  Granger DN. Oxidative stress and cardiac disease. Am J Med 2000; 109:315-323
8. Marttila R, Lorentz H, Rinne U. Oxygen toxicity protecting enzymes in Parkinson’s disease: increase of superoxide dismutase-like activity in the substantia nigra and basal nucleus. Journal of the neurological sciences 1988; 86:321-331
9. Andersen PM. Amyotrophic lateral sclerosis associated with mutations in the CuZn superoxide dismutase gene. Curr Neurol Neurosci Rep 2006; 6:37-46
10. Ceballos-Picot I, Nicole A, Briand P, Grimber G, Delacourte A, Defossez A, et al. Neuronal-specific expression of human copper-zinc superoxide dismutase gene in transgenic mice: animal model of gene dosage effects in Down’s syndrome. Brain Res 1991; 552:198-214
11. Gilgun-Sherki Y, Melamed E, Offen D. The role of oxidative stress in the pathogenesis of multiple sclerosis: the need for effective antioxidant therapy. J Neurol 2004; 251:261-268
12. Tabner BJ, Turnbull S, El-Agnaf O, Allsop D. Production of reactive oxygen species from aggregating proteins implicated in Alzheimer’s disease, Parkinson’s disease and other neurodegenerative diseases. Curr Top Med Chem 2001; 1:507-517
13. Halliwell B. Free radicals and antioxidants - quo vadis? Trends Pharmacol Sci 2011; 32:125-130
14. Johnson F, Giulivi C. Superoxide dismutases and their impact upon human health. Mol Aspects Med 2005; 26:340-352
15. Miller AF. Superoxide dismutases: active sites that save, but a protein that kills. Curr Opin Chem Biol 2004; 8:162-168
16. Noori-Zadeh A, Mesbah-Namin SA, Tiraihi T, Rajabibazl M, Taheri T. Non-viral human proGDNF gene delivery to rat bone marrow stromal cells under ex vivo conditions. J Neurol Sci 2014; 339:81-86
17. Garcı́a Ro, Aguiar J, Alberti E, de la Cuétara K, Pavón N. Bone marrow stromal cells produce nerve growth factor and glial cell line-derived neurotrophic factors. Biochem Biophys Res Commun 2004; 316: 753-754
18. Wu Y, Zheng Q, Xie Z, Wang Y, Hao J, Liu X. Expression of brain-derived neurotrophic factor and nerve growth factor in bone marrow mesenchymal stem cells and therapeutic effect in spinal cord injury. Chin J Exp Surg 2004; 22:139-141
19. Taghi GM, Maryam GK, Taghi L, Leili H, Leyla M. Characterization of in vitro cultured bone marrow and adipose tissue‐derived mesenchymal stem cells and their ability to express neurotrophic factors. Cell Biol Int 2012; 36:1239-1249
20. Marcus DL, Thomas C, Rodriguez C, Simberkoff K, Tsai JS, Strafaci JA, et al. Increased peroxidation and reduced antioxidant enzyme activity in Alzheimer’s disease. Exp Neurol 1998; 150:40-44
21. Nakao N, Frodl EM, Widner H, Carlson E, Eggerding FA, Epstein CJ, et al. Overexpressing Cu/Zn superoxide dismutase enhances survival of transplanted neurons in a rat model of Parkinson’s disease. Nat Med 1995; 1:226-231
22. Browne SE, Bowling AC, Macgarvey U, Baik MJ, Berger SC, Muquit MM, et al. Oxidative damage and metabolic dysfunction in Huntington’s disease: selective vulnerability of the basal ganglia. Ann Neurol 1997; 41:646-653
23. Ohtsuki T, Matsumoto M, Suzuki K, Taniguchi N  Kamada T. Effect of transient forebrain ischemia on superoxide dismutases in gerbil hippocampus. Brain Res 1993; 620:305-309
24. Kinouchi H, Epstein CJ, Mizui T, Carlson E, Chen SF  Chan PH. Attenuation of focal cerebral ischemic injury in transgenic mice overexpressing CuZn superoxide dismutase. Proc Natl Acad Sci U S A 1991; 88:11158-11162
25. Rothstein JD, Bristol LA, Hosler B, Brown RH, Kuncl RW. Chronic inhibition of superoxide dismutase produces apoptotic death of spinal neurons. Proc Natl Acad Sci U S A 1994; 91:4155-4159
26. Greenlund LJ, Deckwerth TL, Johnson Jr EM. Superoxide dismutase delays neuronal apoptosis: a role for reactive oxygen species in programmed neuronal death. Neuron 1995; 14:303-315
27. Troy CM, Shelanski ML. Down-regulation of copper/zinc superoxide dismutase causes apoptotic death in PC12 neuronal cells. P Proc Natl Acad Sci U S A 1994; 91:6384-6387
28. Aminbakhsh A, Mancini J. Chronic antioxidant use and changes in endothelial dysfunction: a review of clinical investigations. Can J Cardiol 1999; 15:895-903
29. Ripps ME, Huntley GW, Hof PR, Morrison JH, Gordon JW. Transgenic mice expressing an altered murine superoxide dismutase gene provide an animal model of amyotrophic lateral sclerosis. Proc Natl Acad Sci U S A 1995; 92:689-693
30. Deitch JS, Alexander GM, Bensinger A, Yang S, Jiang JT, Heiman-Patterson TD. Phenotype of transgenic mice carrying a very low copy number of the mutant human G93A superoxide dismutase-1 gene associated with amyotrophic lateral sclerosis. PLoS One 2014; 9: e99879
31. Ferraiuolo L, Higginbottom A, Heath PR, Barber S, Greenald D, Kirby J,  et al. Dysregulation of astrocyte-motoneuron cross-talk in mutant superoxide dismutase 1-related amyotrophic lateral sclerosis. Brain 2011; 134:2627-2641
32. Lin D, Barnett M, Grauer L, Robben J, Jewell A, Takemoto L, et al. Expression of superoxide dismutase in whole lens prevents cataract formation. Mol Vis 2005; 11:853-858
33. Foresman EL, Miller FJ Jr. Extracellular but not cytosolic superoxide dismutase protects against oxidant-mediated endothelial dysfunction. Redox Biol 2013; 1:292-296.