VEGF delivery with retrogradely transported lentivector prolongs survival in a mouse ALS model


Amyotrophic lateral sclerosis (ALS) causes adult-onset, progressive motor neuron degeneration in the brain and spinal cord, resulting in paralysis and death three to five years after onset in most patients1. ALS is still incurable, in part because its complex aetiology remains insufficiently understood. Recent reports have indicated that reduced levels of vascular endothelial growth factor (VEGF), which is essential in angiogenesis and has also been implicated in neuroprotection2,3,4, predispose mice and humans to ALS5,6. However, the therapeutic potential of VEGF for the treatment of ALS has not previously been assessed. Here we report that a single injection of a VEGF-expressing lentiviral vector into various muscles delayed onset and slowed progression of ALS in mice engineered to overexpress the gene coding for the mutated G93A form of the superoxide dismutase-1 (SOD1G93A) (refs 7–10), even when treatment was only initiated at the onset of paralysis. VEGF treatment increased the life expectancy of ALS mice by 30 per cent without causing toxic side effects, thereby achieving one of the most effective therapies reported in the field so far.

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Figure 1: EIAV vector-mediated gene transfer in vitro and in vivo.
Figure 2: Effects of EIAV vector delivery of VEGF.
Figure 3: VEGF gene therapy protects spinal and brainstem motor neurons in SOD1G93A transgenic mice.
Figure 4: The effects of VEGF gene therapy five weeks after intramuscular injections of viral vectors.


  1. 1

    Mulder, D. W. Clinical limits of amyotrophic lateral sclerosis. Adv. Neurol. 36, 15–22 (1982)

    CAS  PubMed  Google Scholar 

  2. 2

    Carmeliet, P. Blood vessels and nerves: common signals, pathways and diseases. Nature Rev. Genet. 4, 710–720 (2003)

    CAS  Article  Google Scholar 

  3. 3

    Schratzberger, P. et al. Favorable effect of VEGF gene transfer on ischemic peripheral neuropathy. Nature Med. 6, 405–413 (2000)

    CAS  Article  Google Scholar 

  4. 4

    Jin, K. L., Mao, X. O. & Greenberg, D. A. Vascular endothelial growth factor: direct neuroprotective effect in in vitro ischemia. Proc. Natl Acad. Sci. USA 97, 10242–10247 (2000)

    ADS  CAS  Article  Google Scholar 

  5. 5

    Oosthuyse, B. et al. Deletion of the hypoxia-response element in the vascular endothelial growth factor promoter causes motor neuron degeneration. Nature Genet. 28, 131–138 (2001)

    CAS  Article  Google Scholar 

  6. 6

    Lambrechts, D. et al. VEGF is a modifier of amyotrophic lateral sclerosis in mice and humans and protects motoneurons against ischemic death. Nature Genet. 34, 383–394 (2003)

    CAS  Article  Google Scholar 

  7. 7

    Gurney, M. E. et al. Motor neuron degeneration in mice that express a human Cu,Zn superoxide dismutase mutation. Science 264, 1772–1775 (1994)

    ADS  CAS  Article  Google Scholar 

  8. 8

    Bruijn, L. I. et al. ALS-linked SOD1 mutant G85R mediates damage to astrocytes and promotes rapidly progressive disease with SOD1-containing inclusions. Neuron 18, 327–338 (1997)

    CAS  Article  Google Scholar 

  9. 9

    Azzouz, M. et al. Increased motoneuron survival and improved neuromuscular function in transgenic ALS mice after intraspinal injection of an adeno-associated virus encoding Bcl-2. Hum. Mol. Genet. 9, 803–811 (2000)

    CAS  Article  Google Scholar 

  10. 10

    Cleveland, D. W. & Rothstein, J. D. From Charcot to Lou Gehrig: deciphering selective motor neuron death in ALS. Nature Rev. Neurosci. 2, 806–819 (2001)

    CAS  Article  Google Scholar 

  11. 11

    Mazarakis, N. D. et al. Rabies virus glycoprotein pseudotyping of lentiviral vectors enables retrograde axonal transport and access to the nervous system after peripheral delivery. Hum. Mol. Genet. 10, 2109–2121 (2001)

    CAS  Article  Google Scholar 

  12. 12

    Wong, L. F. et al. Transduction patterns of pseudotyped lentiviral vectors in the nervous system. Mol. Ther. 9, 101–111 (2004)

    CAS  Article  Google Scholar 

  13. 13

    Rosen, D. R. Mutations in Cu/Zn superoxide dismutase gene are associated with familial amyotrophic lateral sclerosis. Nature 364, 362 (1993)

    ADS  CAS  Article  Google Scholar 

  14. 14

    Ferreirinha, F. et al. Axonal degeneration in paraplegin-deficient mice is associated with abnormal mitochondria and impairment of axonal transport. J. Clin. Invest. 113, 231–242 (2004)

    CAS  Article  Google Scholar 

  15. 15

    Sagot, Y., Rosse, T., Vejsada, R., Perrelet, D. & Kato, A. C. Differential effects of neurotrophic factors on motoneuron retrograde labeling in a murine model of motoneuron disease. J. Neurosci. 18, 1132–1141 (1998)

    CAS  Article  Google Scholar 

  16. 16

    Dal Canto, M. C. & Gurney, M. E. Neuropathological changes in two lines of mice carrying a transgene for mutant human Cu,Zn SOD, and in mice overexpressing wild type human SOD: a model of familial amyotrophic lateral sclerosis (FALS). Brain Res. 676, 25–40 (1995)

    CAS  Article  Google Scholar 

  17. 17

    Wang, L. J. et al. Neuroprotective effects of glial cell line-derived neurotrophic factor mediated by an adeno-associated virus vector in a transgenic animal model of amyotrophic lateral sclerosis. J. Neurosci. 22, 6920–6928 (2002)

    CAS  Article  Google Scholar 

  18. 18

    Kaspar, B., Llado, J., Sherkat, N., Rothstein, J. & Gage, F. Retrograde viral delivery of IGF-1 prolongs survival in a mouse ALS model. Science 301, 839–842 (2003)

    ADS  CAS  Article  Google Scholar 

  19. 19

    Fukuda, R. et al. Insulin-like growth factor 1 induces hypoxia-inducible factor 1-mediated vascular endothelial growth factor expression, which is dependent on MAP kinase and phosphatidylinositol 3-kinase signaling in colon cancer cells. J. Biol. Chem. 277, 38205–38211 (2002)

    CAS  Article  Google Scholar 

  20. 20

    Hellstrom, A. et al. Low IGF-I suppresses VEGF-survival signaling in retinal endothelial cells: direct correlation with clinical retinopathy of prematurity. Proc. Natl Acad. Sci. USA 98, 5804–5808 (2001)

    ADS  CAS  Article  Google Scholar 

  21. 21

    Mitchell, J. D., Wokke, J. H. & Borasio, G. D. Recombinant human insulin-like growth factor I (rhIGF-I) for amyotrophic lateral sclerosis/motor neuron disease. Cochrane Database Syst. Rev. 3, CD002064 (2002)

    Google Scholar 

  22. 22

    Azzouz, M. et al. Multicistronic lentiviral vector-mediated striatal gene transfer of aromatic L-amino acid decarboxylase, tyrosine hydroxylase, and GTP cyclohydrolase I induces sustained transgene expression, dopamine production, and functional improvement in a rat model of Parkinson's disease. J. Neurosci. 22, 10302–10312 (2002)

    CAS  Article  Google Scholar 

  23. 23

    Bienemann, A. S. et al. Long-term replacement of a mutated nonfunctional CNS gene: reversal of hypothalamic diabetes insipidus using an EIAV-based lentiviral vector expressing arginine vasopressin. Mol. Ther. 7, 588–596 (2003)

    CAS  Article  Google Scholar 

  24. 24

    Mitrophanous, K. et al. Stable gene transfer to the nervous system using a non-primate lentiviral vector. Gene Ther. 6, 1808–1818 (1999)

    CAS  Article  Google Scholar 

  25. 25

    Rohll, J. B. et al. Design, production, safety, evaluation, and clinical applications of nonprimate lentiviral vectors. Methods Enzymol. 346, 466–500 (2002)

    CAS  Article  Google Scholar 

  26. 26

    Martin-Rendon, E., White, L. J., Olsen, A., Mitrophanous, K. A. & Mazarakis, N. D. New methods to titrate EIAV-based lentiviral vectors. Mol. Ther. 5, 566–570 (2002)

    CAS  Article  Google Scholar 

  27. 27

    Mohammed, S. I. et al. Effects of the cyclooxygenase inhibitor, piroxicam, on tumor response, apoptosis, and angiogenesis in a canine model of human invasive urinary bladder cancer. Cancer Res. 62, 356–358 (2002)

    CAS  PubMed  Google Scholar 

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We thank H. Mehmet and A. Kingsman for critical review of the manuscript. SOD1G93A transgenic mice were a gift from P. Shaw. Oxford BioMedica supported this work.

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Correspondence to Mimoun Azzouz or Nicholas D. Mazarakis.

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Some authors in this paper hold shares at Oxford Biomedica (Ltd).

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EIAV-VEGF gene therapy in SOD1G93A mice induce minimal immune response (PPT 651 kb)

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Azzouz, M., Ralph, G., Storkebaum, E. et al. VEGF delivery with retrogradely transported lentivector prolongs survival in a mouse ALS model. Nature 429, 413–417 (2004).

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