Cargando…

Gene Transfer of Mutant Mouse Cholinesterase Provides High Lifetime Expression and Reduced Cocaine Responses with No Evident Toxicity

Gene transfer of a human cocaine hydrolase (hCocH) derived from butyrylcholinesterase (BChE) by 5 mutations (A199S/F227A/S287G/A328W/Y332G) has shown promise in animal studies for treatment of cocaine addiction. To predict the physiological fate and immunogenicity of this enzyme in humans, a compara...

Descripción completa

Detalles Bibliográficos
Autores principales: Geng, Liyi, Gao, Yang, Chen, Xiabin, Hou, Shurong, Zhan, Chang-Guo, Radic, Zoran, Parks, Robin J., Russell, Stephen J., Pham, Linh, Brimijoin, Stephen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3696080/
https://www.ncbi.nlm.nih.gov/pubmed/23840704
http://dx.doi.org/10.1371/journal.pone.0067446
_version_ 1782476283918155776
author Geng, Liyi
Gao, Yang
Chen, Xiabin
Hou, Shurong
Zhan, Chang-Guo
Radic, Zoran
Parks, Robin J.
Russell, Stephen J.
Pham, Linh
Brimijoin, Stephen
author_facet Geng, Liyi
Gao, Yang
Chen, Xiabin
Hou, Shurong
Zhan, Chang-Guo
Radic, Zoran
Parks, Robin J.
Russell, Stephen J.
Pham, Linh
Brimijoin, Stephen
author_sort Geng, Liyi
collection PubMed
description Gene transfer of a human cocaine hydrolase (hCocH) derived from butyrylcholinesterase (BChE) by 5 mutations (A199S/F227A/S287G/A328W/Y332G) has shown promise in animal studies for treatment of cocaine addiction. To predict the physiological fate and immunogenicity of this enzyme in humans, a comparable enzyme was created and tested in a conspecific host. Thus, similar mutations (A199S/S227A/S287G/A328W/Y332G) were introduced into mouse BChE to obtain a mouse CocH (mCocH). The cDNA was incorporated into viral vectors based on: a) serotype-5 helper-dependent adenovirus (hdAD) with ApoE promoter, and b) serotype-8 adeno-associated virus with CMV promoter (AAV-CMV) or multiple promoter and enhancer elements (AAV-VIP). Experiments on substrate kinetics of purified mCocH expressed in HEK293T cells showed 30-fold higher activity (U/mg) with (3)H-cocaine and 25% lower activity with butyrylthiocholine, compared with wild type BChE. In mice given modest doses of AAV-CMV-mCocH vector (0.7 or 3×10(11) particles) plasma hydrolase activity rose 10-fold above control for over one year with no observed immune response. Under the same conditions, transduction of the human counterpart continued less than 2 months and antibodies to hCocH were readily detected. The advanced AAV-VIP-mCocH vector generated a dose-dependent rise in plasma cocaine hydrolase activity from 20-fold (10(10) particles) to 20,000 fold (10(13) particles), while the hdAD vector (1.7×10(12) particles) yielded a 300,000-fold increase. Neither vector caused adverse reactions such as motor weakness, elevated liver enzymes, or disturbance in spontaneous activity. Furthermore, treatment with high dose hdAD-ApoE-mCocH vector (1.7×10(12) particles) prevented locomotor abnormalities, other behavioral signs, and release of hepatic alanine amino transferase after a cocaine dose fatal to most control mice (120 mg/kg). This outcome suggests that viral gene transfer can yield clinically effective cocaine hydrolase expression for lengthy periods without immune reactions or cholinergic dysfunction, while blocking toxicity from drug overdose.
format Online
Article
Text
id pubmed-3696080
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-36960802013-07-09 Gene Transfer of Mutant Mouse Cholinesterase Provides High Lifetime Expression and Reduced Cocaine Responses with No Evident Toxicity Geng, Liyi Gao, Yang Chen, Xiabin Hou, Shurong Zhan, Chang-Guo Radic, Zoran Parks, Robin J. Russell, Stephen J. Pham, Linh Brimijoin, Stephen PLoS One Research Article Gene transfer of a human cocaine hydrolase (hCocH) derived from butyrylcholinesterase (BChE) by 5 mutations (A199S/F227A/S287G/A328W/Y332G) has shown promise in animal studies for treatment of cocaine addiction. To predict the physiological fate and immunogenicity of this enzyme in humans, a comparable enzyme was created and tested in a conspecific host. Thus, similar mutations (A199S/S227A/S287G/A328W/Y332G) were introduced into mouse BChE to obtain a mouse CocH (mCocH). The cDNA was incorporated into viral vectors based on: a) serotype-5 helper-dependent adenovirus (hdAD) with ApoE promoter, and b) serotype-8 adeno-associated virus with CMV promoter (AAV-CMV) or multiple promoter and enhancer elements (AAV-VIP). Experiments on substrate kinetics of purified mCocH expressed in HEK293T cells showed 30-fold higher activity (U/mg) with (3)H-cocaine and 25% lower activity with butyrylthiocholine, compared with wild type BChE. In mice given modest doses of AAV-CMV-mCocH vector (0.7 or 3×10(11) particles) plasma hydrolase activity rose 10-fold above control for over one year with no observed immune response. Under the same conditions, transduction of the human counterpart continued less than 2 months and antibodies to hCocH were readily detected. The advanced AAV-VIP-mCocH vector generated a dose-dependent rise in plasma cocaine hydrolase activity from 20-fold (10(10) particles) to 20,000 fold (10(13) particles), while the hdAD vector (1.7×10(12) particles) yielded a 300,000-fold increase. Neither vector caused adverse reactions such as motor weakness, elevated liver enzymes, or disturbance in spontaneous activity. Furthermore, treatment with high dose hdAD-ApoE-mCocH vector (1.7×10(12) particles) prevented locomotor abnormalities, other behavioral signs, and release of hepatic alanine amino transferase after a cocaine dose fatal to most control mice (120 mg/kg). This outcome suggests that viral gene transfer can yield clinically effective cocaine hydrolase expression for lengthy periods without immune reactions or cholinergic dysfunction, while blocking toxicity from drug overdose. Public Library of Science 2013-06-28 /pmc/articles/PMC3696080/ /pubmed/23840704 http://dx.doi.org/10.1371/journal.pone.0067446 Text en © 2013 Geng et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Geng, Liyi
Gao, Yang
Chen, Xiabin
Hou, Shurong
Zhan, Chang-Guo
Radic, Zoran
Parks, Robin J.
Russell, Stephen J.
Pham, Linh
Brimijoin, Stephen
Gene Transfer of Mutant Mouse Cholinesterase Provides High Lifetime Expression and Reduced Cocaine Responses with No Evident Toxicity
title Gene Transfer of Mutant Mouse Cholinesterase Provides High Lifetime Expression and Reduced Cocaine Responses with No Evident Toxicity
title_full Gene Transfer of Mutant Mouse Cholinesterase Provides High Lifetime Expression and Reduced Cocaine Responses with No Evident Toxicity
title_fullStr Gene Transfer of Mutant Mouse Cholinesterase Provides High Lifetime Expression and Reduced Cocaine Responses with No Evident Toxicity
title_full_unstemmed Gene Transfer of Mutant Mouse Cholinesterase Provides High Lifetime Expression and Reduced Cocaine Responses with No Evident Toxicity
title_short Gene Transfer of Mutant Mouse Cholinesterase Provides High Lifetime Expression and Reduced Cocaine Responses with No Evident Toxicity
title_sort gene transfer of mutant mouse cholinesterase provides high lifetime expression and reduced cocaine responses with no evident toxicity
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3696080/
https://www.ncbi.nlm.nih.gov/pubmed/23840704
http://dx.doi.org/10.1371/journal.pone.0067446
work_keys_str_mv AT gengliyi genetransferofmutantmousecholinesteraseprovideshighlifetimeexpressionandreducedcocaineresponseswithnoevidenttoxicity
AT gaoyang genetransferofmutantmousecholinesteraseprovideshighlifetimeexpressionandreducedcocaineresponseswithnoevidenttoxicity
AT chenxiabin genetransferofmutantmousecholinesteraseprovideshighlifetimeexpressionandreducedcocaineresponseswithnoevidenttoxicity
AT houshurong genetransferofmutantmousecholinesteraseprovideshighlifetimeexpressionandreducedcocaineresponseswithnoevidenttoxicity
AT zhanchangguo genetransferofmutantmousecholinesteraseprovideshighlifetimeexpressionandreducedcocaineresponseswithnoevidenttoxicity
AT radiczoran genetransferofmutantmousecholinesteraseprovideshighlifetimeexpressionandreducedcocaineresponseswithnoevidenttoxicity
AT parksrobinj genetransferofmutantmousecholinesteraseprovideshighlifetimeexpressionandreducedcocaineresponseswithnoevidenttoxicity
AT russellstephenj genetransferofmutantmousecholinesteraseprovideshighlifetimeexpressionandreducedcocaineresponseswithnoevidenttoxicity
AT phamlinh genetransferofmutantmousecholinesteraseprovideshighlifetimeexpressionandreducedcocaineresponseswithnoevidenttoxicity
AT brimijoinstephen genetransferofmutantmousecholinesteraseprovideshighlifetimeexpressionandreducedcocaineresponseswithnoevidenttoxicity