Lentiviral gene transfer of GDNF to the nigrostriatal system in the rat brain

Lentiviral gene transfer of GDNF to the nigrostriatal system in the rat brain

Field: Disorders of the nervous system

Authors: Georgievska, Biljana
Rosenblad, Carl
Lundberg, Cecilia
Kirik, Deniz
Björklund, Anders

Address of presenting
author: Wallenberg Neuroscience Center, Division of Neurobiology, Sölvegatan 17, 223 62 Lund

E-mail: biljana.georgievska@mphy.lu.se

Phone: 046-222 0527

Fax: 046-222 0561

Text of abstract: Recombinant lentiviral (rLV) vectors are attractive tools for gene therapy to the nervous system, since they infect non-dividing cells and allow efficient integration and stable expression of transgenes into terminally differentiated cells. rLV vectors show high transduction efficiency of striatal as well as midbrain cells and are therefore suitable for gene delivery to the nigrostriatal system, the major system affected in Parkinson’s disease. In an initial study, rLV vectors expressing the marker gene GFP and the trophic factor GDNF were injected in the striatum and substantia nigra (Rosenblad et al., MCN in press). The rLV vectors were produced according to Zufferey et al.,1997, and had a titer of 10(7) TU/ml. GFP-expressing cells were found in a 0.7-1.5 mm wide and 2-3 mm deep column at both the striatal and midbrain injection sites. The majority of transduced cells in the striatum had a neuronal morphology. In contrast, many more astroglial cells were transduced in the midbrain while the transduction of cells in SN pars compacta was very low. Immunostaining for GDNF revealed a widespread and diffuse distribution in the striatum and the SN indicating that GDNF diffuses well. In a second experiment the injection parameters for striatal rLV-GDNF transduction were optimised to achieve neuroprotection of nigral dopamine neurons against a terminal 6-OHDA lesion. The vector was diluted 1:2 and 1:10 of which 2 ul was injected in the striatum and 1:50 of which 10 ul was injected. TH+ nigral neurons were spared in a dose-dependent fashion with a maximum protection of 78% of the intact side in the group receiving the 1:2 dilution of rLV-GDNF. These results indicate that the rLV vector system is highly effective for delivery of therapeutic factors to the nigrostriatal system.

Keywords: lentiviral vectors, gene transfer, GDNF, Parkinsons's disease, neuroprotection

	Lentiviral gene transfer of GDNF to the nigrostriatal system in the rat brain
Field:	Disorders of the nervous system
Authors:	Georgievska, Biljana Rosenblad, Carl Lundberg, Cecilia Kirik, Deniz Björklund, Anders
Address of presenting author:	Wallenberg Neuroscience Center, Division of Neurobiology, Sölvegatan 17, 223 62 Lund
E-mail:	biljana.georgievska@mphy.lu.se
Phone:	046-222 0527
Fax:	046-222 0561
Text of abstract:	Recombinant lentiviral (rLV) vectors are attractive tools for gene therapy to the nervous system, since they infect non-dividing cells and allow efficient integration and stable expression of transgenes into terminally differentiated cells. rLV vectors show high transduction efficiency of striatal as well as midbrain cells and are therefore suitable for gene delivery to the nigrostriatal system, the major system affected in Parkinson’s disease. In an initial study, rLV vectors expressing the marker gene GFP and the trophic factor GDNF were injected in the striatum and substantia nigra (Rosenblad et al., MCN in press). The rLV vectors were produced according to Zufferey et al.,1997, and had a titer of 10(7) TU/ml. GFP-expressing cells were found in a 0.7-1.5 mm wide and 2-3 mm deep column at both the striatal and midbrain injection sites. The majority of transduced cells in the striatum had a neuronal morphology. In contrast, many more astroglial cells were transduced in the midbrain while the transduction of cells in SN pars compacta was very low. Immunostaining for GDNF revealed a widespread and diffuse distribution in the striatum and the SN indicating that GDNF diffuses well. In a second experiment the injection parameters for striatal rLV-GDNF transduction were optimised to achieve neuroprotection of nigral dopamine neurons against a terminal 6-OHDA lesion. The vector was diluted 1:2 and 1:10 of which 2 ul was injected in the striatum and 1:50 of which 10 ul was injected. TH+ nigral neurons were spared in a dose-dependent fashion with a maximum protection of 78% of the intact side in the group receiving the 1:2 dilution of rLV-GDNF. These results indicate that the rLV vector system is highly effective for delivery of therapeutic factors to the nigrostriatal system.

Keywords:	lentiviral vectors, gene transfer, GDNF, Parkinsons's disease, neuroprotection

Index

Created 2000-03-13

Department of Physiological Sciences, Lund University

Web programming FormOnLine AB