Targeted retrograde gene delivery for neuronal protection.

The cellular heterogeneity and complex circuitry of the central nervous system make it difficult to achieve precise delivery of experimental and therapeutic agents. We report here an in vivo retrograde gene delivery strategy to target mature projection neurons using adeno-associated virus, a vector with low toxicity and the capacity for long-term gene expression. Viral delivery to axon terminal fields in the hippocampus and striatum resulted in viral internalization, retrograde transport, and transgene expression in specific projection neurons in entorhinal cortex and substantia nigra. Retrograde delivery of the anti-apoptotic gene Bcl2l (also known as Bcl-xL) protected entorhinal projection neurons from subsequent damage-induced cell death. Given the broad distribution of neurons affected by neurodegenerative diseases, gene delivery to both the terminal fields and the projection neurons through retrograde infection provides for strategic therapeutic intervention at both levels of the neural circuit. This approach may also facilitate experimental studies of defined neural circuits.

[1]  J. Qian,et al.  Behavioral and Cellular Protection of Rat Dopaminergic Neurons by an Adenoviral Vector Encoding Glial Cell Line-Derived Neurotrophic Factor , 1998, Experimental Neurology.

[2]  D. Cleveland.  From Charcot to SOD1 Mechanisms of Selective Motor Neuron Death in ALS , 1999, Neuron.

[3]  N. L. Chamberlin,et al.  Recombinant adeno-associated virus vector: use for transgene expression and anterograde tract tracing in the CNS , 1998, Brain Research.

[4]  F. Gage,et al.  Perforant path damage results in progressive neuronal death and somal atrophy in layer II of entorhinal cortex and functional impairment with increasing postdamage age , 1994, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[5]  D. Bohl,et al.  Efficient and stable adeno-associated virus-mediated transduction in the skeletal muscle of adult immunocompetent mice. , 1997, Human gene therapy.

[6]  K. Kissa,et al.  Preferential transduction of neurons by canine adenovirus vectors and their efficient retrograde transport in vivo , 2001, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[7]  I. Martins,et al.  Recombinant adeno-associated virus type 2, 4, and 5 vectors: transduction of variant cell types and regions in the mammalian central nervous system. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[8]  M. J. Wade,et al.  Neuron number in the entorhinal cortex and CA1 in preclinical Alzheimer disease. , 2001, Archives of neurology.

[9]  R. Duvoisin.  Overview of Parkinson's Disease , 1992, Annals of the New York Academy of Sciences.

[10]  M. King,et al.  Neuron-Specific Transduction in the Rat Septohippocampal or Nigrostriatal Pathway by Recombinant Adeno-associated Virus Vectors , 1998, Experimental Neurology.

[11]  D A Peterson,et al.  Quantitative histology using confocal microscopy: implementation of unbiased stereology procedures. , 1999, Methods.

[12]  R. Samulski,et al.  AAV vectors: is clinical success on the horizon? , 2000, Gene Therapy.

[13]  D. Bennett,et al.  Loss and atrophy of layer II entorhinal cortex neurons in elderly people with mild cognitive impairment , 2001, Annals of neurology.

[14]  R. Samulski,et al.  Infectious Entry Pathway of Adeno-Associated Virus and Adeno-Associated Virus Vectors , 2000, Journal of Virology.

[15]  W. Cowan,et al.  A quantitative EM autoradiographic study of the commissural and associational connections of the dentate gyrus in the rat , 2004, Anatomy and Embryology.

[16]  M. Bobinski,et al.  Prediction of cognitive decline in normal elderly subjects with 2-[18F]fluoro-2-deoxy-d-glucose/positron-emission tomography (FDG/PET) , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[17]  J. Morris,et al.  Profound Loss of Layer II Entorhinal Cortex Neurons Occurs in Very Mild Alzheimer’s Disease , 1996, The Journal of Neuroscience.

[18]  B. Davidson,et al.  Transduction of murine cerebellar neurons with recombinant FIV and AAV5 vectors , 2000, Neuroreport.

[19]  N. Kawai,et al.  Adeno-associated virus vector-mediated bcl-2 gene transfer into post-ischemic gerbil brain in vivo: prospects for gene therapy of ischemia-induced neuronal death , 2000, Gene Therapy.

[20]  Keyun Qing,et al.  Human fibroblast growth factor receptor 1 is a co-receptor for infection by adeno-associated virus 2 , 1999, Nature Medicine.

[21]  R. Samulski,et al.  Fluorescent viral vectors: A new technique for the pharmacological analysis of gene therapy , 1998, Nature Medicine.

[22]  Absence of Helper Adenovirus Adeno-Associated Virus Vectors in the Production of High-Titer Recombinant , 1998 .

[23]  J. Glorioso,et al.  Herpes simplex virus vector-mediated expression of Bcl-2 prevents 6-hydroxydopamine-induced degeneration of neurons in the substantia nigra in vivo. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[24]  F. Gage,et al.  Future Prospects of Gene Therapy for Treating CNS Diseases , 2000 .

[25]  Donald W. Pfaff,et al.  Long-term gene expression and phenotypic correction using adeno-associated virus vectors in the mammalian brain , 1994, Nature Genetics.

[26]  J. Friedman,et al.  Virus-Assisted Mapping of Neural Inputs to a Feeding Center in the Hypothalamus , 2001, Science.

[27]  A. Holtmaat,et al.  Transient gene transfer to neurons and glia: Analysis of adenoviral vector performance in the CNS and PNS , 1997, Journal of Neuroscience Methods.

[28]  C. Thompson,et al.  bcl-x is expressed in embryonic and postnatal neural tissues and functions to prevent neuronal cell death. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[29]  F. Gage,et al.  Central neuronal loss and behavioral impairment in mice lacking neurotrophin receptor p75. , 1999, The Journal of comparative neurology.

[30]  X. Breakefield,et al.  Herpes simplex virus for gene delivery to neurons. , 1991, The New biologist.

[31]  R. Samulski,et al.  Selective and rapid uptake of adeno-associated virus type 2 in brain. , 1998, Human gene therapy.

[32]  D. Amaral,et al.  Entorhinal cortex of the rat: Topographic organization of the cells of origin of the perforant path projection to the dentate gyrus , 1998, The Journal of comparative neurology.

[33]  F. Gage,et al.  Bcl-xL protects adult septal cholinergic neurons from axotomized cell death. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[34]  Juan Li,et al.  Gene Transfer by Adeno-Associated Virus Vectors into the Central Nervous System , 1997, Experimental Neurology.

[35]  R. Samulski,et al.  Viral receptors and vector purification: New approaches for generating clinical-grade reagents , 1999, Nature Medicine.

[36]  James N. Davis,et al.  BCL-2 Transduction, Using a Herpes Simplex Virus Amplicon, Protects Hippocampal Neurons from Transient Global Ischemia , 1999, Experimental Neurology.

[37]  D. Peterson,et al.  Fibroblast growth factor-2 protects entorhinal layer II glutamatergic neurons from axotomy-induced death , 1996, The Journal of neuroscience : the official journal of the Society for Neuroscience.