In Silico Screening for Angiogenesis-Related Genes in Rat Astrocytes

  • Kim, Soo-Young (Division of Pharmaceutical Bioscience, College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, Graduate Program in Bioinformatics, Seoul National University) ;
  • Lee, Sae-Won (Division of Pharmaceutical Bioscience, College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University) ;
  • You, Sung Yong (Division of Pharmaceutical Bioscience, College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University) ;
  • Rha, Sun Young (Cancer Metastasis Research Center, Yonsei University College of Medicine) ;
  • Kim, Kyu-Won (Division of Pharmaceutical Bioscience, College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University)
  • Published : 2004.03.01

Abstract

Astrocytes play supportive roles for neurons in the brain. Recently, they have been accepted to have various functions in the vascular system as well as in the nervous system. We investigated the differential gene expression in rat astrocytes according to the oxygen tension, which is a crucial factor for angiogenesis. A cDNA microarray was performed to find the genes whose expression was sensitive to oxygen tension. We found 26 genes in the astrocyte were found and classified into 4 groups. In order to show the genes' relevancy to angiogenesis, seven of the 26 genes were investigated to see whether they have capabilities of interaction with angiogenesis­related factors in AngioDB. Through this investigation, we found interactions of three proteins with angiogenesis-related factors. These genes were further investigated with a new focus on the vascular endothelial growth factor (VEGF) expression in an astrocyte based on our hypothesis that astrocytes can have effects on endothelial angiogenesis via the release of VEGF. Collectively, we identified several genes whose expressions were dependent on the oxygen concentration of the astrocyte. Furthermore, the relevancy of astrocytes to angiogenesis was investigated using preexisting information of AngioDB, and suggested a possible signaling pathway for VEGF expression in the aspects of brain endothelial angiogenesis by astrocytes.

Keywords

References

  1. Benezra R, Rafii S, and Lyden D (2001). The Id proteins and angiogenesis. Oncogene 20, 8334-8341 https://doi.org/10.1038/sj.onc.1205160
  2. Dentelli P, Del Sorbo L, Rosso A, Molinar A, Garbarino G, Camussi G, Pegoraro L, and Brizzi MF (1999). Human IL-3 stimulates endothelial cell motility and promotes in vivo new vessel formation. J. Immunol. 163, 2151-2159
  3. Derynck R, Zhang Y, and FengXH (1998). Smads:transcriptional activators of TGF-beta responses. Cell 95, 737-40 https://doi.org/10.1016/S0092-8674(00)81696-7
  4. Di Cristofano A, Niki M, Zhao M, Kamell FG, Clarkson B, Pear WS, Van Aelst L, and Pandolfi PP (2001). p62(dok), a negative regulator of Ras and mitogen-activated protein kinase(MAPK) activity, opposes leukemogenesis by p21O(bcr-abl). J. Exp. Med. 194, 275-284 https://doi.org/10.1084/jem.194.3.275
  5. Howe JR, Roth S, Ringold JC, Summers RW, Jarvinen HJ, Sistonen P, Tomlinson IP, Houlston RS, Bevan S, Mitros FA, Stone EM, and Aaltonen LA (1998). Mutations in the SMAD4/DPC4 gene in juvenile polyposis. Science 280, 1086-1088 https://doi.org/10.1126/science.280.5366.1086
  6. Janzer RC and Raft MC (1987), Astrocytes induce blood-brain barrier properties in endothelial cells. Nature 325, 253-257 https://doi.org/10.1038/325253a0
  7. Lee SW, Kim WJ, Choi YK, Song HS, Son MJ, Gelman IH, Kim YJ, and Kim KW (2003). SSeCKS regulates angiogenesis and tight junction formation in blood-brain barrier. Nat. Med. 9, 900-906 https://doi.org/10.1038/nm889
  8. Lu D, Yang H, and Raizada MK (1998). Involvement of p62 nucleoporin in angiotensin II-induced nuclear translocation of STAT3 in brain neurons. J. Neurosci. 18, 1329-1336
  9. Maltepe E and Simon MC(1998). Oxygen, genes, and devlopment: an analysis of the role of hypoxic gene regulation during murine vascular development. J. Mol. Med. 76, 391-401 https://doi.org/10.1007/s001090050231
  10. Pal S, Datta K, Khosravi-Far R, and Mukhopadhyay D (2001). Role of protein kinase Czeta in Ras-mediated transcriptional activation of vascular permeability factor/vascular endothelial growth factor expression. J. Biol. Chem. 276, 2395-2403 https://doi.org/10.1074/jbc.M007818200
  11. Pardali K, Kurisaki A, Moren A, ten Dijke P, Kardassis D, and Moustakas A (2000). Role of Smad proteins and transcription factor Sp1 in p21(Waf1/Cip1) regulation by transforming growth factor-beta. J. Biol. Chem. 275, 29244-29256 https://doi.org/10.1074/jbc.M909467199
  12. Park I, Chung J, Walsh CT, Yun Y, Strominger JL, and Shin J (1995). Phosphotyrosine-independent binding of a 62-kDa protein to the src homology 2(SH2) domain of p56lck and its regulation by phosphorylation of Ser-59 in the Ick unique N-terminal region. Proc. Natl. Acad. Sci. US4 92, 12338-12342 https://doi.org/10.1073/pnas.92.26.12338
  13. Park JA, Choi KS, Kim SY, and Kim KW (2003). Coordinated interaction of the vascular and nervous systems: from molecule-to cell-based approaches. Biochem. Biophys. Res. Commun. 311, 247-253 https://doi.org/10.1016/j.bbrc.2003.09.129
  14. Parri R and Crunelli V (2003). An astrocyte bridge from synapse to blood flow. Nat. Neurosci. 6. 5-6 https://doi.org/10.1038/nn0103-5
  15. Puis A, Schmidt S, Grawe F, and Stabel S (1997). Interaction of protein kinase C zeta with ZIP, a novel protein kinase C-binding protein. Proc. Natl. Acad. Sci. USA 94, 6191-6196 https://doi.org/10.1073/pnas.94.12.6191
  16. Reisinger K, Kaufmann R, and Gille J (2003). Increased Sp1 phosphorylation as a mechanism of hepatocyte growth factor (HGF/SF)-induced vascular endothelial growth factor(VEGF/ VPF) transcription. J. Cell. Sci. 116, 225-238 https://doi.org/10.1242/jcs.00237
  17. Rizkalla B, Forbes JM, Cooper ME, and Cao Z (2003). Increased renal vascular endothelial growth factor and angiopoietins by angiotensin II infusion is mediated by both AT1 and AT2 receptors. J. Am. Soc. Nephrol. 14, 3061-3071 https://doi.org/10.1097/01.ASN.0000099374.58607.C9
  18. Roberts AB, Sporn MB, Assoian RK, Smith JM, Roche NS, Wakefield LM, Heine UI.,Liotla LA, Falanga V, and Kehrl JH (1986). Transforming growth factor type : rapid induction of fibrosis and angiogenesis in vivcand stimulation of collagen formation in vitro. Proc. Natl. Acad. Sci. USA 83, 4167-4171 https://doi.org/10.1073/pnas.83.12.4167
  19. Schafer G, Cramer T, Suske G, Kemmner W, Wiedenmann B, and Hocker M (2003). Oxidative stress regulates vascular endothelial growth factor-A gene transcription through Sp1- and Sp3-dependent activation of two proximal GC-rich promoter elements. J. Biol. Chem. 278, 8190-8198 https://doi.org/10.1074/jbc.M211999200
  20. Shih AY, Johnson DA, Wong G, Kraft AD, Jiang L, Erb H, Johnson JA, and Murphy TH (2003). Coordinate regulation of glutathione biosynthesis and release by Nrf2-expressing glia potently protects neurons from oxidative stress. J. Neurosci. 23, 3394-406
  21. Sohn TK, Moon EJ, Lee SK, Coo HG, and Kim KW (2002). AngioDB: database of angiogenesis and angiogenesis-related molecules. Nucleic. Acids. Res. 30, 369-71 https://doi.org/10.1093/nar/30.1.369
  22. Travis J (1994), Glia: the brain's other cells. Science 206, 970-922
  23. Yang E, Zha J, Jockel J, Boise LH, Thomson CB, and Korsemeyer SJ (1995), Bad, a heterodimeric partner for Bel-xL and BeI-2, displaces Bax and promotes cell death. Cell 80, 285-291 https://doi.org/10.1016/0092-8674(95)90411-5
  24. Zhu Y, Yang GY, Ahlemeyer B, Pang L, Che XM, Culmsee C, Klumpp S, and Krieglstein J (2002). Transforming growth factor-beta 1 increases bad phosphorylation and protects neurons against damage. J. Neurosci. 22, 3898-3909
  25. Zonta M, Angulo MC, Gobbo S, Rosengarten B, Hossmann KA, Pozzan T, and Carmignoto G. (2003). Neuron-to-astrocyte signaling is central to the dynamic control of brain microcirculation. Nat. Neurosci. 6, 43-50 https://doi.org/10.1038/nn980