Skip to main content
SpringerLink
Log in

Treatment of Focal Brain Ischemia with Viral Vector-Mediated Gene Transfer

  • Protocol
  • First Online:
The Blood-Brain and Other Neural Barriers

Part of the book series: Methods in Molecular Biology ((MIMB,volume 686))

Abstract

Promoting functional recovery after ischemic brain injury has emerged as a potential approach for the treatment of ischemic stroke. An ideal restorative approach to enhance long-term functional recovery is to promote postischemic angiogenesis and neurogenesis. This chapter describes a system using adeno-associated viral (AAV) vector-mediated vascular endothelial growth factor (VEGF) gene transfer into the ischemic brain. The methods described here for construction, production, and purification of AAV vector expressing VEGF gene can also be applied to producing AAV vectors expressing other genes. This chapter also illustrates the methods to produce mouse middle cerebral artery occlusion (MCAO), injection of viral vector into the mouse brain, and standard assays for determining the success of brain ischemia and gene transfer.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 139.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 179.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Weintraub M I (2006) Thrombolysis (tissue plasminogen activator) in stroke: a medicolegal quagmire. Stroke 37, 1917–1922

    Article  PubMed  Google Scholar 

  2. Ohab JJ, Fleming S, Blesch A, Carmichael ST (2006) A neurovascular niche for neurogenesis after stroke. J Neurosci 26, 13007–13016

    Article  PubMed  CAS  Google Scholar 

  3. del Zoppo GJ (2006) Stroke and neurovascular protection. N Engl J Med 354, 553–555

    Article  PubMed  Google Scholar 

  4. McCown TJ (2005) Adeno-associated virus (AAV) vectors in the CNS. Curr Gene Ther 5, 333–338

    Article  PubMed  CAS  Google Scholar 

  5. Leker RR, Soldner F, Velasco I, Gavin DK, Androutsellis-Theotokis A, McKay RD (2007) Long-lasting regeneration after ischemia in the cerebral cortex. Stroke 38, 153–161

    Article  PubMed  Google Scholar 

  6. Shen F, Su H, Liu W, Kan YW, Young WL, Yang GY (2006) Recombinant adeno-associated viral vector encoding human VEGF165 induces neomicrovessel formation in the adult mouse brain. Front Biosci 11, 3190–3198

    Article  PubMed  CAS  Google Scholar 

  7. Sun Y, Jin K, Xie L, Childs J, Mao XO, Logvinova A, Greenberg DA (2003) VEGF-induced neuroprotection, neurogenesis, and angiogenesis after focal cerebral ischemia. J Clin Invest 111, 1843–1851

    PubMed  CAS  Google Scholar 

  8. Bellomo M, Adamo EB, Deodato B, Catania MA, Mannucci C, Marini H, Marciano MC, Marini R, Sapienza S, Giacca M, Caputi AP, Squadrito F, Calapai G (2003) Enhancement of expression of vascular endothelial growth factor after adeno-associated virus gene transfer is associated with improvement of brain ischemia injury in the gerbil. Pharmacol Res 48, 309–317

    Article  PubMed  CAS  Google Scholar 

  9. Andsberg G, Kokaia Z, Klein RL, Muzyczka N, Lindvall O, Mandel R J (2002) Neuropathological and behavioral consequences of adeno-associated viral vector-mediated continuous intrastriatal neurotrophin delivery in a focal ischemia model in rats. Neurobiol Dis 9, 187–204

    Article  PubMed  CAS  Google Scholar 

  10. Tsai TH, Chen SL, Chiang YH, Lin SZ, Ma H I, Kuo SW, Tsao YP (2000) Recombinant adeno-associated virus vector expressing glial cell line-derived neurotrophic factor reduces ischemia-induced damage. Exp Neurol 166, 266–275

    Article  PubMed  CAS  Google Scholar 

  11. Xiao W, Chirmule N, Berta SC, McCullough B, Gao G, Wilson JM (1999) Gene therapy vectors based on adeno-associated virus type 1. J Virol 73, 3994–4003

    PubMed  CAS  Google Scholar 

  12. Weber M, Rabinowitz J, Provost N, Conrath H, Folliot S, Briot D, Cherel Y, Chenuaud P, Samulski J, Moullier P, Rolling F (2003) Recombinant adeno-associated virus serotype 4 mediates unique and exclusive long-term transduction of retinal pigmented epithelium in rat, dog, and nonhuman primate after subretinal delivery. Mol Ther 7, 774–781

    Article  PubMed  CAS  Google Scholar 

  13. Zabner J, Seiler M, Walters R, Kotin RM, Fulgeras W, Davidson BL, Chiorini JA (2000) Adeno-associated virus type 5 (AAV5) but not AAV2 binds to the apical surfaces of airway epithelia and facilitates gene transfer. J Virol 74, 3852–3858

    Article  PubMed  CAS  Google Scholar 

  14. Davidson BL, Stein CS, Heth JA, Martins I, Kotin RM, Derksen TA, Zabner J, Ghodsi A, Chiorini JA (2000) Recombinant adeno-associated virus type 2, 4, and 5 vectors: transduction of variant cell types and regions in the mammalian central nervous system. Proc Natl Acad Sci U S A 97, 3428–3432

    Article  PubMed  CAS  Google Scholar 

  15. Chao H, Liu Y, Rabinowitz J, Li C, Samulski RJ, Walsh CE (2000) Several log increase in therapeutic transgene delivery by distinct adeno-associated viral serotype vectors. Mol Ther 2, 619–623

    Article  PubMed  CAS  Google Scholar 

  16. Chao H, Monahan PE, Liu Y, Samulski RJ, Walsh CE (2001) Sustained and complete phenotype correction of hemophilia B mice following intramuscular injection of AAV1 serotype vectors. Mol Ther 4, 217–222

    Article  PubMed  CAS  Google Scholar 

  17. Duan D, Yan Z, Yue Y, Ding W, Engelhardt JF (2001) Enhancement of muscle gene delivery with pseudotyped adeno-associated virus type 5 correlates with myoblast differentiation. J Virol 75, 7662–7671

    Article  PubMed  CAS  Google Scholar 

  18. Braddock M (2005) Safely slowing down the decline in Alzheimer’s disease: gene therapy shows potential. Expert Opin Investig Drugs 14, 913–915

    Article  PubMed  CAS  Google Scholar 

  19. Tuszynski MH, Thal L, Pay M, Salmon DP, U HS, Bakay R, Patel P, Blesch A, Vahlsing HL, Ho G, Tong G, Potkin SG, Fallon J, Hansen L, Mufson EJ, Kordower JH, Gall C, Conner J (2005) A phase 1 clinical trial of nerve growth factor gene therapy for Alzheimer disease. Nat Med 11, 551–555

    Article  PubMed  CAS  Google Scholar 

  20. McPhee SW, Janson CG, Li C, Samulski RJ, Camp AS, Francis J, Shera D, Lioutermann L, Feely M, Freese A, Leone P (2006) Immune responses to AAV in a phase I study for Canavan disease. J Gene Med 8, 577–588

    Article  PubMed  CAS  Google Scholar 

  21. Fulci G, Chiocca EA (2007) The status of gene therapy for brain tumors. Expert Opin Biol Ther 7, 197–208

    Article  PubMed  CAS  Google Scholar 

  22. Colombo F, Barzon L, Franchin E, Pacenti M, Pinna V, Danieli D, Zanusso M, Palu G (2005) Combined HSV-TK/IL-2 gene therapy in patients with recurrent glioblastoma multiforme: biological and clinical results. Cancer Gene Ther 12, 835–848

    Article  PubMed  CAS  Google Scholar 

  23. Ren H, Boulikas T, Lundstrom K, Soling A, Warnke PC, Rainov NG (2003) Immunogene therapy of recurrent glioblastoma multiforme with a liposomally encapsulated replication-incompetent Semliki forest virus vector carrying the human interleukin-12 gene–a phase I/II clinical protocol. J Neurooncol 64, 147–154

    PubMed  CAS  Google Scholar 

  24. Rainov NG (2000) A phase III clinical evaluation of herpes simplex virus type 1 thymidine kinase and ganciclovir gene therapy as an adjuvant to surgical resection and radiation in adults with previously untreated glioblastoma multiforme. Hum Gene Ther 11, 2389–2401

    Article  PubMed  CAS  Google Scholar 

  25. Chu Y, Miller JD, Heistad DD (2007) Gene therapy for stroke: 2006 overview. Curr Hypertens Rep 9, 19–24

    Article  PubMed  CAS  Google Scholar 

  26. Heistad DD (2006) Gene therapy for vascular disease. Vascul Pharmacol 45, 331–333

    Article  PubMed  CAS  Google Scholar 

  27. Hsich G, Sena-Esteves M, Breakefield XO (2002) Critical issues in gene therapy for neurologic disease. Hum Gene Ther 13, 579–604

    Article  PubMed  CAS  Google Scholar 

  28. Yenari MA, Dumas TC, Sapolsky RM, Steinberg GK (2001) Gene therapy for treatment of cerebral ischemia using defective herpes simplex viral vectors. Ann N Y Acad Sci 939, 340–357

    Article  PubMed  CAS  Google Scholar 

  29. Watanabe T, Okuda Y, Nonoguchi N, Zhao M Z, Kajimoto Y, Furutama D, Yukawa H, Shibata MA, Otsuki Y, Kuroiwa T, Miyatake S (2004) Postischemic intraventricular administration of FGF-2 expressing adenoviral vectors improves neurologic outcome and reduces infarct volume after transient focal cerebral ischemia in rats. J Cereb Blood Flow Metab 24, 1205–1213

    Article  PubMed  CAS  Google Scholar 

  30. Shimamura M, Sato N, Oshima K, Aoki M, Kurinami H, Waguri S, Uchiyama,Y, Ogihara T, Kaneda Y, Morishita R (2004) Novel therapeutic strategy to treat brain ischemia: overexpression of hepatocyte growth factor gene reduced ischemic injury without cerebral edema in rat model. Circulation 109, 424–431

    Article  PubMed  CAS  Google Scholar 

  31. Li H, Qian ZM (2002) Transferrin/transferrin receptor-mediated drug delivery. Med Res Rev 22, 225–250

    Article  PubMed  CAS  Google Scholar 

  32. Pang L, Ye W, Che XM, Roessler BJ, Betz A L, Yang GY (2001) Reduction of inflammatory response in the mouse brain with adenoviral- mediated transforming growth factor-ss1 expression. Stroke 32, 544–552

    PubMed  CAS  Google Scholar 

  33. Hayashi K, Morishita R, Nakagami H, Yoshimura S, Hara A, Matsumoto K, Nakamura T, Ogihara T, Kaneda Y, Sakai N (2001) Gene therapy for preventing neuronal death using hepatocyte growth factor: in vivo gene transfer of HGF to subarachnoid space prevents delayed neuronal death in gerbil hippocampal CA1 neurons. Gene Ther 8, 1167–1173

    Article  PubMed  CAS  Google Scholar 

  34. Yang GY, Mao Y, Zhou LF, Ye W, Liu XH, Gong C, Betz AL (1999) Attenuation of temporary focal cerebral ischemic injury in the mouse following transfection with interleukin-1 receptor antagonist. Brain Res Mol Brain Res 72, 129–137

    Article  PubMed  CAS  Google Scholar 

  35. Betz AL, Yang GY, Davidson BL (1995) Attenuation of stroke size in rats using an adenoviral vector to induce overexpression of interleukin-1 receptor antagonist in brain. J Cereb Blood Flow Metab 15, 547–551

    PubMed  CAS  Google Scholar 

  36. Davis AS, Zhao H, Sun GH, Sapolsky RM, Steinberg GK (2007) Gene therapy using SOD1 protects striatal neurons from experimental stroke. Neurosci Lett 411, 32–36

    Article  PubMed  CAS  Google Scholar 

  37. Badin RA, Lythgoe MF, van der Weerd L, Thomas DL, Gadian DG, Latchman DS (2006) Neuroprotective effects of virally delivered HSPs in experimental stroke. J Cereb Blood Flow Metab 26, 371–381

    Article  PubMed  CAS  Google Scholar 

  38. Baker AH, Sica V, Work LM, Williams-Ignarro S, de Nigris F, Lerman LO, Casamassimi A, Lanza A, Schiano C, Rienzo M, Ignarro LJ, Napoli C (2007) Brain protection using autologous bone marrow cell, metalloproteinase inhibitors, and metabolic treatment in cerebral ischemia. Proc Natl Acad Sci U S A 104, 3597–3602

    Article  PubMed  CAS  Google Scholar 

  39. Frampton AR Jr, Goins WF, Nakano K, Burton EA, Glorioso JC (2005) HSV trafficking and development of gene therapy vectors with applications in the nervous system. Gene Ther 12, 891–901

    Article  PubMed  CAS  Google Scholar 

  40. Wang C, Wang CM, Clark KR, Sferra TJ (2003) Recombinant AAV serotype 1 transduction efficiency and tropism in the murine brain. Gene Ther 10, 1528–1534

    Article  PubMed  CAS  Google Scholar 

  41. Xu DG, Crocker SJ, Doucet JP, St-Jean M, Tamai K, Hakim AM, Ikeda JE, Liston P, Thompson CS, Korneluk RG, MacKenzie A, Robertson GS (1997) Elevation of neuronal expression of NAIP reduces ischemic damage in the rat hippocampus. Nat Med 3, 997–1004

    Article  PubMed  CAS  Google Scholar 

  42. Pechan PA, Yoshida T, Panahian N, Moskowitz MA, Breakefield XO (1995) Genetically modified fibroblasts producing NGF protect hippocampal neurons after ischemia in the rat. Neuroreport 6, 669–672

    Article  PubMed  CAS  Google Scholar 

  43. Miao HS, Yu LY, Hui GZ, Guo LH (2005) Antiapoptotic effect both in vivo and in vitro of A20 gene when transfected into rat hippocampal neurons. Acta Pharmacol Sin 26, 33–38

    Article  PubMed  CAS  Google Scholar 

  44. Shirakura M, Fukumura M, Inoue M, Fujikawa S, Maeda M, Watabe K, Kyuwa S, Yoshikawa Y, Hasegawa M (2003) Sendai virus vector-mediated gene transfer of glial cell line-derived neurotrophic factor prevents delayed neuronal death after transient global ischemia in gerbils. Exp Anim 52, 119–127

    Article  PubMed  CAS  Google Scholar 

  45. Shen F, Su H, Fan Y-F, Zhu Y, Chen Y, Kan YW, Young WL, Yang GY (2006) Induction of focal angiogenesis through adeno-associated viral vector mediated VEGF165 gene transfer in the mature mouse brain (Abstract). Stroke 37, 685

    Google Scholar 

  46. Kurozumi K, Nakamura K, Tamiya T, Kawano Y, Ishii K, Kobune M, Hirai S, Uchida H, Sasaki K, Ito Y, Kato K, Honmou O, Houkin K, Date I, Hamada H (2005) Mesenchymal stem cells that produce neurotrophic factors reduce ischemic damage in the rat middle cerebral artery occlusion model. Mol Ther 11, 96–104

    Article  PubMed  CAS  Google Scholar 

  47. Gu W, Zhao H, Yenari MA, Sapolsky R M, Steinberg GK (2004) Catalase over-expression protects striatal neurons from transient focal cerebral ischemia. Neuroreport 15, 413–416

    Article  PubMed  CAS  Google Scholar 

  48. Hoehn B, Yenari MA, Sapolsky RM, Steinberg GK (2003) Glutathione peroxidase overexpression inhibits cytochrome C release and proapoptotic mediators to protect neurons from experimental stroke. Stroke 34, 2489–2494

    Article  PubMed  CAS  Google Scholar 

  49. Tsai TH, Chen SL, Xiao X, Chiang YH, Lin SZ, Kuo SW, Liu DW, Tsao YP (2003) Gene treatment of cerebral stroke by rAAV vector delivering IL-1ra in a rat model. Neuroreport 14, 803–807

    Article  PubMed  CAS  Google Scholar 

  50. Lawrence M S, McLaughlin JR, Sun GH, Ho DY, McIntosh L, Kunis DM, Sapolsky R M, Steinberg GK (1997) Herpes simplex viral vectors expressing Bcl-2 are neuroprotective when delivered after a stroke. J Cereb Blood Flow Metab 17, 740–744

    Article  PubMed  CAS  Google Scholar 

  51. Zhao MZ, Nonoguchi N, Ikeda N, Watanabe T, Furutama D, Miyazawa D, Funakoshi H, Kajimoto Y, Nakamura T, Dezawa M, Shibata MA, Otsuki Y, Coffin RS, Liu WD, Kuroiwa T, Miyatake S (2006) Novel therapeutic strategy for stroke in rats by bone marrow stromal cells and ex vivo HGF gene transfer with HSV-1 vector. J Cereb Blood Flow Metab 26, 1176–1188

    Article  PubMed  CAS  Google Scholar 

  52. Sugiura S, Kitagawa K, Tanaka S, Todo K, Omura-Matsuoka E, Sasaki T, Mabuchi T, Matsushita K, Yagita Y, Hori M (2005) Adenovirus-mediated gene transfer of heparin-binding epidermal growth factor-like growth factor enhances neurogenesis and angiogenesis after focal cerebral ischemia in rats. Stroke 36, 859–864

    Article  PubMed  CAS  Google Scholar 

  53. Sun Y, Jin K, Clark KR, Peel A, Mao XO, Chang Q, Simon RP, Greenberg DA (2003) Adeno-associated virus-mediated delivery of BCL-w gene improves outcome after transient focal cerebral ischemia. Gene Ther 10, 115–122

    Article  PubMed  CAS  Google Scholar 

  54. Hadaczek P, Kohutnicka M, Krauze MT, Bringas J, Pivirotto P, Cunningham J, Bankiewicz, K (2006) Convection-enhanced delivery of adeno-associated virus type 2 (AAV2) into the striatum and transport of AAV2 within monkey brain. Hum Gene Ther 17, 291–302

    Article  PubMed  CAS  Google Scholar 

  55. Vilaboa N, Voellmy R (2006) Regulatable gene expression systems for gene therapy. Curr Gene Ther 6, 421–438

    Article  PubMed  CAS  Google Scholar 

  56. Yang GY, Zhao Y, Davidson BL, Betz AL (1997) Overexpression of interleukin-1 receptor antagonist in the mouse brain reduces ischemic brain injury. Brain Res 751, 181–188

    Article  PubMed  CAS  Google Scholar 

  57. Yang GY, Xu B, Hashimoto T, Huey M, Chaly T, Jr, Wen R, Young WL (2003) Induction of focal angiogenesis through adenoviral vector mediated vascular endothelial cell growth factor gene transfer in the mature mouse brain. Angiogenesis 6, 151–158

    Article  PubMed  CAS  Google Scholar 

  58. Lee CZ, Xu B, Hashimoto T, McCulloch CE, Yang GY, Young WL (2004) Doxycycline suppresses cerebral matrix metalloproteinase-9 and angiogenesis induced by focal hyperstimulation of vascular endothelial growth factor in a mouse model. Stroke 35, 1715–1719

    Article  PubMed  CAS  Google Scholar 

  59. Gao G P, Alvira MR, Wang L, Calcedo R, Johnston J, Wilson JM (2002) Novel adeno-associated viruses from rhesus monkeys as vectors for human gene therapy. Proc Natl Acad Sci U S A 99, 11854–11859

    Article  PubMed  CAS  Google Scholar 

  60. Rutledge EA, Halbert CL, Russell DW (1998) Infectious clones and vectors derived from adeno-associated virus (AAV) serotypes other than AAV type 2. J Virol 72, 309–319

    PubMed  CAS  Google Scholar 

  61. Muramatsu S, Mizukami H, Young NS, Brown K E (1996) Nucleotide sequencing and generation of an infectious clone of adeno-associated virus 3. Virology 221, 208–217

    Article  PubMed  CAS  Google Scholar 

  62. Chiorini JA, Yang L, Liu Y, Safer B, Kotin RM (1997) Cloning of adeno-associated virus type 4 (AAV4) and generation of recombinant AAV4 particles. J Virol 71, 6823–6833

    PubMed  CAS  Google Scholar 

  63. Chiorini JA, Kim F, Yang L, Kotin RM (1999) Cloning and characterization of adeno-associated virus type 5. J Virol 73, 1309–1319

    PubMed  CAS  Google Scholar 

  64. Du L, Sullivan CC, Chu D, Cho AJ, Kido M, Wolf PL, Yuan JX, Deutsch R, Jamieson S W, Thistlethwaite PA (2003) Signaling molecules in nonfamilial pulmonary hypertension. N Engl J Med 348, 500–509

    Article  PubMed  CAS  Google Scholar 

  65. Clark KR, Liu X, McGrath JP, Johnson PR (1999) Highly purified recombinant adeno-associated virus vectors are biologically active and free of detectable helper and wild-type viruses. Hum Gene Ther 10, 1031–1039

    Article  PubMed  CAS  Google Scholar 

  66. Zolotukhin S, Potter M, Zolotukhin I, Sakai Y, Loiler S, Fraites,TJ Jr, Chiodo VA, Phillipsberg T, Muzyczka N, Hauswirth WW, Flotte TR, Byrne BJ, Snyder RO (2002) Production and purification of serotype 1, 2, and 5 recombinant adeno-associated viral vectors. Methods 28, 158–167

    Article  PubMed  CAS  Google Scholar 

  67. Zolotukhin S, Byrne BJ, Mason E, Zolotukhin I, Potter M, Chesnut K, Summerford C, Samulski RJ, Muzyczka N (1999) Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield. Gene Ther 6, 973–985

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Center for Cerebrovascular Research, Department of Anesthesia and Perioperative Care, University of California San Francisco, San Francisco, CA, USA

    Hua Su & Guo-Yuan Yang

  2. Med-X Research Institute, Sanghai JiaoTong University, Shanghai, China

    Guo-Yuan Yang

Authors
  1. Hua Su
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Guo-Yuan Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Banting Institute, Dept. Laboratory Med. & Pathobiology, University of Toronto, College St. 100, Toronto, M5G 1L5, Ontario, Canada

    Sukriti Nag

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Su, H., Yang, GY. (2011). Treatment of Focal Brain Ischemia with Viral Vector-Mediated Gene Transfer. In: Nag, S. (eds) The Blood-Brain and Other Neural Barriers. Methods in Molecular Biology, vol 686. Humana Press. https://doi.org/10.1007/978-1-60761-938-3_22

Download citation

  • DOI: https://doi.org/10.1007/978-1-60761-938-3_22

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-937-6

  • Online ISBN: 978-1-60761-938-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation