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Optimization of continuous in vivo DOPA production and studies on ectopic DA synthesis using rAAV5 vectors in Parkinsonian rats

Björklund, Tomas LU ; Hall, Helene LU ; Breysse, Nathalie LU ; Soneson, Charlotte LU ; Carlsson, Thomas LU ; Mandel, Ronald J.; Carta, Manolo LU and Kirik, Deniz LU (2009) In Journal of Neurochemistry 111(2). p.355-367
Abstract
Viral vector-mediated gene transfer is emerging as a novel therapeutic approach with clinical utility in treatment of Parkinson's disease. Recombinant adeno-associated viral (rAAV) vector in particular has been utilized for continuous l-3,4 dihydroxyphenylalanine (DOPA) delivery by expressing the tyrosine hydroxylase (TH) and GTP cyclohydrolase 1 (GCH1) genes which are necessary and sufficient for efficient synthesis of DOPA from dietary tyrosine. The present study was designed to determine the optimal stoichiometric relationship between TH and GCH1 genes for ectopic DOPA production and the cellular machinery involved in its synthesis, storage, and metabolism. For this purpose, we injected a fixed amount of rAAV5-TH vector and increasing... (More)
Viral vector-mediated gene transfer is emerging as a novel therapeutic approach with clinical utility in treatment of Parkinson's disease. Recombinant adeno-associated viral (rAAV) vector in particular has been utilized for continuous l-3,4 dihydroxyphenylalanine (DOPA) delivery by expressing the tyrosine hydroxylase (TH) and GTP cyclohydrolase 1 (GCH1) genes which are necessary and sufficient for efficient synthesis of DOPA from dietary tyrosine. The present study was designed to determine the optimal stoichiometric relationship between TH and GCH1 genes for ectopic DOPA production and the cellular machinery involved in its synthesis, storage, and metabolism. For this purpose, we injected a fixed amount of rAAV5-TH vector and increasing amounts of rAAV5-GCH1 into the striatum of rats with complete unilateral dopamine lesion. After 7 weeks the animals were killed for either biochemical or histological analysis. We show that increasing the availability of 5,6,7,8-tetrahydro-l-biopterin (BH4) in the same cellular compartment as the TH enzyme resulted in better efficiency in DOPA synthesis, most likely by hindering inactivation of the enzyme and increasing its stability. Importantly, the BH4 synthesis from ectopic GCH1 expression was saturable, yielding optimal TH enzyme functionality between GCH1 : TH ratios of 1 : 3 and 1 : 7. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
tyrosine hydroxylase, recombinant adeno-associated viral vector, disease, Parkinson's, GTP cyclohydrolase-1, dopamine, enzyme replacement
in
Journal of Neurochemistry
volume
111
issue
2
pages
355 - 367
publisher
Wiley-Blackwell
external identifiers
  • wos:000270152600007
  • pmid:19686242
  • scopus:70349319755
ISSN
1471-4159
DOI
10.1111/j.1471-4159.2009.06340.x
language
English
LU publication?
yes
id
4f2de3e9-4dc1-4902-a33e-87790a137eb1 (old id 1489983)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/19686242?dopt=Abstract
date added to LUP
2009-10-21 10:26:50
date last changed
2017-03-03 16:31:52
@article{4f2de3e9-4dc1-4902-a33e-87790a137eb1,
  abstract     = {Viral vector-mediated gene transfer is emerging as a novel therapeutic approach with clinical utility in treatment of Parkinson's disease. Recombinant adeno-associated viral (rAAV) vector in particular has been utilized for continuous l-3,4 dihydroxyphenylalanine (DOPA) delivery by expressing the tyrosine hydroxylase (TH) and GTP cyclohydrolase 1 (GCH1) genes which are necessary and sufficient for efficient synthesis of DOPA from dietary tyrosine. The present study was designed to determine the optimal stoichiometric relationship between TH and GCH1 genes for ectopic DOPA production and the cellular machinery involved in its synthesis, storage, and metabolism. For this purpose, we injected a fixed amount of rAAV5-TH vector and increasing amounts of rAAV5-GCH1 into the striatum of rats with complete unilateral dopamine lesion. After 7 weeks the animals were killed for either biochemical or histological analysis. We show that increasing the availability of 5,6,7,8-tetrahydro-l-biopterin (BH4) in the same cellular compartment as the TH enzyme resulted in better efficiency in DOPA synthesis, most likely by hindering inactivation of the enzyme and increasing its stability. Importantly, the BH4 synthesis from ectopic GCH1 expression was saturable, yielding optimal TH enzyme functionality between GCH1 : TH ratios of 1 : 3 and 1 : 7.},
  author       = {Björklund, Tomas and Hall, Helene and Breysse, Nathalie and Soneson, Charlotte and Carlsson, Thomas and Mandel, Ronald J. and Carta, Manolo and Kirik, Deniz},
  issn         = {1471-4159},
  keyword      = {tyrosine hydroxylase,recombinant adeno-associated viral vector,disease,Parkinson's,GTP cyclohydrolase-1,dopamine,enzyme replacement},
  language     = {eng},
  number       = {2},
  pages        = {355--367},
  publisher    = {Wiley-Blackwell},
  series       = {Journal of Neurochemistry},
  title        = {Optimization of continuous in vivo DOPA production and studies on ectopic DA synthesis using rAAV5 vectors in Parkinsonian rats},
  url          = {http://dx.doi.org/10.1111/j.1471-4159.2009.06340.x},
  volume       = {111},
  year         = {2009},
}