All Title Author
Keywords Abstract

PLOS ONE  2009 

Human Neural Stem Cells Genetically Modified to Overexpress Akt1 Provide Neuroprotection and Functional Improvement in Mouse Stroke Model

DOI: 10.1371/journal.pone.0005586

Full-Text   Cite this paper   Add to My Lib

Abstract:

In a previous study, we have shown that human neural stem cells (hNSCs) transplanted in brain of mouse intracerebral hemorrhage (ICH) stroke model selectively migrate to the ICH lesion and induce behavioral recovery. However, low survival rate of grafted hNSCs in the brain precludes long-term therapeutic effect. We hypothesized that hNSCs overexpressing Akt1 transplanted into the lesion site could provide long-term improved survival of hNSCs, and behavioral recovery in mouse ICH model. F3 hNSC was genetically modified with a mouse Akt1 gene using a retroviral vector. F3 hNSCs expressing Akt1 were found to be highly resistant to H2O2-induced cytotoxicity in vitro. Following transplantation in ICH mouse brain, F3.Akt1 hNSCs induced behavioral improvement and significantly increased cell survival (50–100% increase) at 2 and 8 weeks post-transplantation as compared to parental F3 hNSCs. Brain transplantation of hNSCs overexpressing Akt1 in ICH animals provided functional recovery, and survival and differentiation of grafted hNSCs. These results indicate that the F3.Akt1 human NSCs should be a great value as a cellular source for the cellular therapy in animal models of human neurological disorders including ICH.

References

[1]  Gebel JM, Broderick JP (2000) Intracerebral hemorrhage. Neurol Clin 18: 419–438.
[2]  NINDS ICH Workshop Participants (2005) Priorities for clinical research in intracerebral hemorrhage: report from a National Institute of Neurological Disorders and Stroke workshop. Stroke 36: 23–41.
[3]  McKay R (1997) Stem cells in the central nervous system. Science 276: 66–71.
[4]  Gage FH (2000) Mammalian neural stem cells. Science 287: 1433–1438.
[5]  Kim SU (2004) Human neural stem cells genetically modified for brain repair in neurological disorders. Neuropathology 24: 159–171.
[6]  Lindvall O, Kokaia Z (2006) Stem cells for the treatment of neurological disorders. Nature 441: 1094–1096.
[7]  Muller FJ, Snyder EY, Loring JF (2006) Gene therapy: can neural stem cells deliver? Nat Rev Neurosci 7: 75–84.
[8]  Flax JD, Aurora S, Yang C, Simonin C, Wills AM, et al. (1998) Engraftable human neural stem cells respond to developmental cues, replace neurons, and express foreign genes. Nat Biotech 16: 1033–1039.
[9]  Chu K, Kim M, Jeong SW, Kim SU, Yoon BW (2003) Human neural stem cells can migrate, differentiate, and integrate after intravenous transplantation in adult rats with transient forebrain ischemia. Neurosci Lett 343: 129–133.
[10]  Chu K, Kim M, Park KI, Jeong SW, Park HK, et al. (2004) Human neural stem cells improve sensorimotor deficits in the rat brain with experimental focal ischemia. Brain Res 1016: 145–153.
[11]  Chu K, Kim M, Chae SH, Jeong SW, Kang KS, et al. (2004) Distribution and in situ proliferation patterns of intravenously injected immortalized human neural stem cells in rats with focal cerebral ischemia. Neurosci Res 50: 459–465.
[12]  Jeong SW, Chu K, Jung KH, Kim SU, Kim M, et al. (2003) Human neural stem cell transplantation promotes functional recovery in rats with experimental intracerebral hemorrhage. Stroke 34: 2258–2263.
[13]  Kelly S, Bliss TM, Shah AK, Sun GH, Ma M, et al. (2004) Transplanted human fetal neural stem cells survive, migrate and differentiate in ischemic rat cerebral cortex. Proc Nat Acad Sci USA 101: 11839–11844.
[14]  Ishibashi S, Sakaguchi M, Kuroiwa T, Yamasaki M, Kanemura Y, et al. (2004) Human neural stem/progenitor cells, expanded in long-term neurosphere culture, promote functional recovery after focal ischemia in Mongolian gerbils. J Neurosci Res 78: 215–223.
[15]  Lee HJ, Kim KS, Kim EJ, Choi HB, Lee KH, et al. (2007) Brain transplantation of immortalized human neural stem cells promotes functional recovery in mouse intracerebral hemorrhage stroke model. Stem Cells 25: 1204–1212.
[16]  Lee HJ, Kim KS, Park IH, Kim SU (2007) Human neural stem cells over-expressing VEGF provide neuroprotection, angiogenesis and functional recovery in mouse stroke model. PLoS ONE 2/e156: 1–14.
[17]  Lee ST, Chu K, Jung KH, Kim SJ, et al. (2008) Anti-inflammatory mechanism of intravascular neural stem cell transplantation in hemorrhagic stroke. Brain 131: 616–629.
[18]  Kim SU, Park IH, Kim TH, Kim KS, Choi HB, et al. (2006) Brain transplantation of human neural stem cells transduced with tyrosine hydroxylase and GTP cyclohydrolase 1 provides functional improvement in animal models of Parkinson disease. Neuropathology 26: 129–140.
[19]  Ryu JK, Kim J, Hong SH, Choi HB, Kim SU (2004) Proactive transplantation of human neural stem cells blocks neuronal cell death in rat model of Huntington disease. Neurobiol Disease 16: 68–77.
[20]  Lee ST, Chu K, Park JE, Lee K, Kang L, et al. (2005) Intravenous administration of human neural stem cells induces functional recovery in Huntington's disease rat model. Neurosci Res 52: 243–249.
[21]  Hwang DH, Lee HJ, Kim BG, Joo IS, Kim SU (2008) Intrathecal transplantation of human neural stem cells over-expressing VEGF provide behavioral improvement, disease onset delay and survival extension in transgenic ALS mice. Gene Ther. in press.
[22]  Meng XL, Shen JS, Ohashi T, Maeda H, Kim SU, et al. (2003) Brain transplantation of genetically engineered human neural stem cells globally corrects brain lesions in mucopolysacchridosis VII mouse. J Neurosci Res 74: 266–277.
[23]  Dudek H, Datta SR, Franke TF, Birnbaum MJ, Yao R, et al. (1997) Regulation of neuronal survival by the serine-threonine protein kinase Akt. Science 275: 661–665.
[24]  Kennedy SG, Kandel ES, Cross TK, Hay N (1999) Akt/protein kinase B inhibits cell death by preventing the release of cytochrome c from mitochondria. Mol Cell Biol 19: 5800–5810.
[25]  Brunet A, Datta SR, Greenberg ME (2001) Transcription-dependent and -independent control of neuronal survival by the PI3K-Akt signaling pathway. Curr Opin Neurobiol 11: 297–305.
[26]  Franke TF, Hornik CP, Segev L, Shostak GA, Sugimoto C (2003) PI3K/Akt and apoptosis: size matters. Oncogene 22: 8983–8998.
[27]  Matsuzaki H, Tamatani M, Mitsuda N, Namikawa K, Kiyama H, et al. (1999) Activation of Akt kinase inhibits apoptosis and changes in Bcl-2 and Bax expression induced by nitric oxide in primary hippocampal neurons. J Neurochem 73: 2037–2046.
[28]  Yamaguchi A, Tamatani M, Matsuzaki H, Namikawa K, Kiyama H, et al. (2001) Akt activation protects hippocampal neurons from apoptosis by inhibiting transcriptional activity of p53. J Biol Chem 276: 5256–5264.
[29]  Chong ZZ, Kang JQ, Maiese K (2003) Erythropoietin fosters both intrinsic and extrinsic neuronal protection through modulation of microglia, Akt1, Bad, and caspase-mediated pathways. Br J Pharmacol 138: 1107–1118.
[30]  Kim SU (1985) Antigen expression by glial cells grown in culture. J Neuroimmunol 8: 255–282.
[31]  Kim SU, Moretto G, Lee V, Yu RK (1986) Neuroimmunology of gangliosides in human neurons and glial cells in culture. J Neurosci Res 15: 303–321.
[32]  Cho TS, Bae JH, Choi HB, Kim SS, Suh-Kim H, et al. (2002) Human neural stem cells: Electrophysiological properties of voltage gated ion channels. Neuroreport 13: 1447–1452.
[33]  Ryu JK, Choi HB, Hatori K, Heisel RL, Pelech SL, et al. (2003) Adenosine triphosphate induces proliferation of human neural stem cells: Role of calcium and p70 ribosomal protein S6 kinase. J Neurosci Res 72: 352–362.
[34]  Kim SU, Nagai A, Nakagawa E, Choi HB, Bang JH, et al. (2008) Production and characterization of immortal human neural stem cell line with multipotent differentiation property. Methods Mol Biol 438: 103–121.
[35]  West MJ, Slomianka L, Gundersen HJ (1991) Unbiased stereological estimation of the total number of neurons in the subdivisions of the rat hippocampus using the opost-transplantationical fractionator. Anat Rec 231: 482–497.
[36]  Fukunaga K, Kawano T (2003) Akt is a molecular target for signal transduction therapy in brain ischemic insult. J Pharmacol Sci 92: 317–327.
[37]  Cardone MH, Roy N, Stennicke SM (1998) Regulation of cell death protease caspase-9 by phosphorylation. Science 282: 1318–1321.
[38]  Taylor JM, Ali U, Iannello RC, Hertzog P, Crack PC (2005) Diminished Akt phosphorylation in neurons lacking glutathione peroxidase-1 (Gpx1) leads to increased susceptibility to oxidative stress-induced cell death. J Neurochem 92: 283–293.
[39]  Rojo AI, Salinas M, Martin D, Perona R, Cuadrado A (2004) Regulation of Cu/Zn-superoxide dismutase expression via the phosphatidylinositol 3 kinase/Akt pathway and nuclear factor-kB. J Neurosci 24: 7324–7334.
[40]  Salinas M, Wang J, de Sagarra M, Martin D, Rojo A, et al. (2004) Protein kinase Akt/PKB phosphorylates heme oxygenase-1 in vitro and in vivo. FEBS Lett 578: 90–94.
[41]  Graff JR, Konicek BW, Manulty AM, Wang Z, Houck K, et al. (2000) Increased Akt activity contributes to prostate cancer progression by dramatically accelerating prostate tumor growth and diminishing p27Kip1 expression. J Biol Chem 275: 24500–24505.
[42]  Brognard J, Clark AS, Ni Y, Dennis PA (2001) Akt/protein kinase B is constitutively active in non-small cell lung cancer cells and promotes cellular survival and resistance to chemotherapy and radiation. Cancer Res 61: 3986–3997.
[43]  Roy HK, Olusola BF, Clemens DL, Karolski WJ, Rotashak A, et al. (2002) Akt proto-oncogene overexpression is an early event during sporadic colon carcinogenesis. Carcinogenesis 23: 201–205.

Full-Text

comments powered by Disqus