Delivery of human acetylcholinesterase by adeno-associated virus to the acetylcholinesterase knockout mouse

Anna Hrabovska, Ellen G. Duysen, Jeffrey D. Sanders, L. Charles Murrin, Oksana Lockridge

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The purpose of this work was to develop a gene delivery system that expressed acetylcholinesterase (AChE) for prolonged periods. An adeno-associated virus (AAV) expressing human AChE was constructed by cotransfecting three plasmids into HEK 293T cells. The purified vector expressed 0.17 μg AChE per 1 million viral particles in culture medium in 23 h, or 0.8 U/ml. The AAV/hAChE was injected into muscle of adult AChE knockout mice and into the brains of 3-6 week old AChE knockout mice. Intramuscular injection yielded plasma AChE levels approaching 50% of the AChE activity of wild-type mouse plasma. The highest AChE activity was found on day 3 post-injection. AChE activity declined thereafter to a constant 7% of normal. The decreased level was accompanied by the appearance of anti-human AChE antibodies, suggesting partial clearance of AChE from plasma by antibodies. Intrastriatal injection resulted in AChE expression in the striatum. No antibodies were detected in animals treated intrastriatally. Motor coordination was improved and the lifespan of intrastriatally-treated AChE knockout mice was prolonged. Human AChE was expressed in mouse brain for up to 7 months after intrastriatal injection of an AAV/hAChE construct. Gene-therapy to supply AChE to the striatum improved motor coordination and prolonged the life of mice genetically deficient in AChE, probably by reducing their susceptibility to spontaneous seizures. This supports the hypothesis that their seizures are induced by excess acetylcholine.

Original languageEnglish (US)
Pages (from-to)71-78
Number of pages8
JournalChemico-Biological Interactions
Volume157-158
DOIs
StatePublished - Dec 15 2005

Fingerprint

Dependovirus
Acetylcholinesterase
Viruses
Knockout Mice
Plasmas
Injections
Antibodies
Brain
Seizures
Gene therapy
Gene Transfer Techniques
HEK293 Cells
Intramuscular Injections

Keywords

  • Acetylcholinesterase
  • Adeno-associated virus
  • Knockout mouse
  • Seizures

ASJC Scopus subject areas

  • Toxicology

Cite this

Delivery of human acetylcholinesterase by adeno-associated virus to the acetylcholinesterase knockout mouse. / Hrabovska, Anna; Duysen, Ellen G.; Sanders, Jeffrey D.; Murrin, L. Charles; Lockridge, Oksana.

In: Chemico-Biological Interactions, Vol. 157-158, 15.12.2005, p. 71-78.

Research output: Contribution to journalArticle

Hrabovska, Anna ; Duysen, Ellen G. ; Sanders, Jeffrey D. ; Murrin, L. Charles ; Lockridge, Oksana. / Delivery of human acetylcholinesterase by adeno-associated virus to the acetylcholinesterase knockout mouse. In: Chemico-Biological Interactions. 2005 ; Vol. 157-158. pp. 71-78.
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