Protection from the toxicity of diisopropylfluorophosphate by adeno-associated virus expressing acetylcholinesterase.
Organophosphorus esters (OP) are highly toxic chemicals used as pesticides and nerve agents. Their acute toxicity is attributed to inhibition of acetylcholinesterase (AChE, EC 3.1.1.7) in nerve synapses. Our goal was to find a new therapeutic for protection against OP toxicity. We used a gene therapy vector, adeno-associated virus serotype 2 (AAV-2), to deliver murine AChE to AChE-/- mice that have no endogenous AChE activity. The vector encoded the most abundant form of AChE: exons 2, 3, 4, and 6. Two-day old animals, with an immature immune system, were injected. AChE delivered intravenously was expressed up to 5 months in plasma, liver, heart, and lung, at 5-15% of the level in untreated wild-type mice. A few mice formed antibodies, but antibodies did not block AChE activity. The plasma AChE was a mixture of dimers and tetramers. AChE delivered intramuscularly had 40-fold higher activity levels than in wild-type muscle. None of the AChE was collagen-tailed. No retrograde transport through the motor neurons to the central nervous system was detected. AChE delivered intrastriatally assembled into tetramers. In brain, the AAV-2 vector transduced neurons, but not astrocytes and microglia. Vector-treated AChE-/- mice lived longer than saline-treated controls. AChE-/- mice were protected from diisopropylfluorophosphate-induced respiratory failure when the vector was delivered intravenously, but not intrastriatally. Since vector-treated animals had no AChE activity in diaphragm muscle, protection from respiratory failure came from AChE in other tissues. We conclude that AChE scavenged OP and in this way protected the activity of butyrylcholinesterase (BChE, EC 3.1.1.8) in motor endplates.
Li
Bin
B
Eppley Institute, University of Nebraska Medical Center, 986805 Nebraska Medical Center, Omaha, NE 68198-6805, USA.
Duysen
Ellen G
EG
Poluektova
Larisa Y
LY
Murrin
L Charles
LC
Lockridge
Oksana
O
D000110:Acetylcholinesterase / Q000097:blood / Q000235:genetics / Q000378:metabolism*; D000256:Adenoviridae / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000201:enzymology; D002800:Cholinesterase Inhibitors / Q000633:toxicity; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics / Q000493:pharmacokinetics; D007269:Injections, Intra-Arterial; D007273:Injections, Intramuscular; D007275:Injections, Intravenous; D007278:Injections, Spinal; D007531:Isoflurophate / Q000633:toxicity*; D051379:Mice; D018345:Mice, Knockout; D008970:Molecular Weight; D018485:Muscle Fibers, Skeletal / Q000201:enzymology; D018482:Muscle, Skeletal / Q000187:drug effects / Q000503:physiopathology; D009474:Neurons / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D016019:Survival Analysis; D013997:Time Factors
D002800:Cholinesterase Inhibitors; D007531:Isoflurophate; D000110:Acetylcholinesterase