Gene therapy of murine motor neuron disease using adenoviral vectors for neurotrophic factors

Abstract

Motor neuron diseases such as amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy cause progressive paralysis, often leading to premature death. Neurotrophic factors have been suggested as therapeutic agents for motor neuron diseases, but their clinical use as injected recombinant protein was limited by toxicity and/or poor bioavailability. We demonstrate here that adenovirus-mediated gene transfer of neurotrophin-3 (NT-3) can produce substantial therapeutic effects in the mouse mutant pmn (progressive motor neuronopathy). After intramuscular injection of the NT-3 adenoviral vector, pmn mice showed a 50% increase in life span, reduced loss of motor axons and improved neuromuscular function as assessed by electromyography. These results were further improved by coinjecting an adenoviral vector coding for ciliary neurotrophic factor. Therefore, adenovirus-mediated gene transfer of neurotrophic factors offers new prospects for the treatment of motor neuron diseases.

Bcl-2 expression by retrograde transport of adenoviral vectors with Cre-loxP recombination system in motor neurons of mutant SOD1 transgenic mice

“…In their investigation of gene-therapeutic modalities for motor neuron diseases, Aebischer et al 8 used encapsulated, genetically modified xenogeneic cells to deliver ciliary neurotrophic factor (CNTF) intrathecally into patients with ALS. Haase et al 9 demonstrated that adenovirus-mediated gene transfer of neurotrophin-3 (NT-3) or NT-3 and CNTF could produce substantial therapeutic effects in pmn (progressive motor neuropathy) mutant mice after intramuscular injection of the NT-3 or NT-3 and CNTF adenoviral vectors. Azzouz et al 10 demonstrated that local anti-apoptotic protein Bcl-2 expression in the spinal motor neurons could increase motor neuron survival and improve neuromuscular function in FALS mice after intraspinal injection of recombinant adeno-associated virus encoding Bcl-2.…”

A neuron-specific gene transfer by a recombinant defective Sindbis virus

“…neurotrophic factor GDNF , BDNF, or neurotrophin-3 Ž . NT-3 was shown to ameliorate neurodegenerative process induced in brain and motor neurons in vivo w x Ž 6, 15,19,20 . However, accommodations for small size -.…”

Pure populations of transduced primary human cells can be produced using GFP expressing herpes virus vectors and flow cytometry

“…The ability to express multiple genes might be important in many therapeutic situations such as, for example, in motor neuron disease where it has recently been shown that the use of two neurotrophic factor expressing adenoviruses was more effective at preventing motor neuron degeneration in an animal model than was either virus alone. 5,6 Similarly multiple gene expression may be useful where a marker for transfected cells is required together with delivery of a 'test' gene in studies of gene function. Alternatively, as we show here, the marker gene can be used for the purification of cells transduced ex vivo to homogeneity when combined with flow cytometry.…”