Dose-dependent neuroprotective effect of ciliary neurotrophic factor delivered via tetracycline-regulated lentiviral vectors in the quinolinic acid rat model of Huntington's disease
The ability to regulate gene expression constitutes a prerequisite for the development of gene therapy strategies aimed at the treatment of neurologic disorders. In the present work, we used tetracycline (Tet)-regulated lentiviral vectors to investigate the dose-dependent neuroprotective effect of human ciliary neurotrophic factor (CNTF) in the quinolinic acid (QA) model of Huntington's disease (HD). The Tet system was split in two lentiviruses, the first one containing the CNTF or green fluorescent protein (GFP) cDNAs under the control of the Tet-response element (TRE) and a second vector encoding the transactivator (tTA). Preliminary coinfection study demonstrated that 63.8% +/- 2.0% of infected cells contain at least two viral copies. Adult rats were then injected with CNTF- and GFP-expressing viral vectors followed 3 weeks later by an intrastriatal administration of QA. A significant reduction of apomorphine-induced rotations was observed in the CNTF-on group. In contrast, GFP-treated animals or CNTF-off rats displayed an ipsilateral turning behavior in response to apomorphine. A selective sparing of DARPP-32-, choline acetyltransferase (ChAT)-, and NADPH-d-positive neurons was observed in the striatum of CNTF-on rats compared to GFP animals and CNTF-off group. Enzyme-linked immunosorbent assay (ELISA) performed on striatal samples of rats sacrificed at the same time point indicated that this neuroprotective effect was associated with the production of 15.5 +/- 4.7 ng CNTF per milligram of protein whereas the residual CNTF expression in the off state (0.54 +/- 0.02 ng/mg of protein) was not sufficient to protect against QA toxicity. These results establish the proof of principle of neurotrophic factor dosing for neurodegenerative diseases and demonstrate the feasibility of lentiviral-mediated tetracycline-regulated gene transfer in the brain.
Keywords: Animals ; Brain/pathology ; Choline O-Acetyltransferase/metabolism ; Ciliary Neurotrophic Factor/ genetics/ pharmacology ; DNA ; Complementary/metabolism ; Disease Models ; Animal ; Dose-Response Relationship ; Drug ; Enzyme-Linked Immunosorbent Assay ; Female ; Genetic Vectors ; Green Fluorescent Proteins ; Humans ; Huntington Disease/ prevention & control/ therapy ; Lentivirus/ genetics ; Luminescent Proteins/metabolism ; Neurons/cytology/metabolism ; Quinolinic Acid/ pharmacology ; Rats ; Rats ; Wistar ; Recombinant Fusion Proteins/metabolism ; Tetracycline/ pharmacology ; Animal ; Rats
Institute of Neurosciences, Swiss Federal Institute of Technology Lausanne, 1015 Lausanne, Switzerland.
Record created on 2007-03-09, modified on 2016-08-08