PMID- 22607388
OWN - NLM
STAT- MEDLINE
DCOM- 20121008
LR  - 20151119
IS  - 1742-4658 (Electronic)
IS  - 1742-464X (Linking)
VI  - 279
IP  - 14
DP  - 2012 Jul
TI  - Targeted delivery into motor nerve terminals of inhibitors for SNARE-cleaving 
      proteases via liposomes coupled to an atoxic botulinum neurotoxin.
PG  - 2555-67
LID - 10.1111/j.1742-4658.2012.08638.x [doi]
AB  - A targeted drug carrier (TDC) is described for transferring functional proteins 
      or peptides into motor nerve terminals, a pivotal locus for therapeutics to treat 
      neuromuscular disorders. It exploits the pronounced selectivity of botulinum 
      neurotoxin type B (BoNT/B) for interacting with acceptors on these cholinergic 
      nerve endings and becoming internalized. The gene encoding an innocuous BoNT/B 
      protease-inactive mutant (BoTIM) was fused to that for core streptavidin, 
      expressed in Escherichia coli and the purified protein was conjugated to 
      surface-biotinylated liposomes. Such decorated liposomes, loaded with fluorescein 
      as traceable cargo, acquired pronounced specificity for motor nerve terminals in 
      isolated mouse hemidiaphragms and facilitated the intraneuronal transfer of the 
      fluor, as revealed by confocal microscopy. Delivery of the protease light chain 
      of botulinum neurotoxin type A (BoNT/A) via this TDC accelerated the onset of 
      neuromuscular paralysis, indicative of improved translocation of this enzyme into 
      the presynaptic cytosol with subsequent proteolytic inactivation of 
      synaptosomal-associated protein of molecular mass 25 kDa (SNAP-25), an exocytotic 
      soluble N-ethyl-maleimide-sensitive factor attachment protein receptor (SNARE) 
      essential for neurotransmitter release. BoTIM-coupled liposomes, loaded with 
      peptide inhibitors of proteases, yielded considerable attenuation of the 
      neuroparalytic effects of BoNT/A or BoNT/F as a result of their cytosolic 
      transfer, the first in situ demonstration of the ability of designer 
      antiproteases to suppress the symptoms of botulism ex vivo. Delivery of the 
      BoNT/A inhibitor by liposomes targeted with the full-length BoTIM proved more 
      effective than that mediated by its C-terminal neuroacceptor-binding domain. This 
      demonstrated versatility of TDC for nonviral cargo transfer into cholinergic 
      nerve endings has unveiled its potential for direct delivery of functional 
      targets into motor nerve endings.
CI  - (c) 2012 The Authors Journal compilation (c) 2012 FEBS.
FAU - Edupuganti, Om P
AU  - Edupuganti OP
AD  - International Centre for Neurotherapeutics, Dublin City University, Dublin, 
      Ireland.
FAU - Ovsepian, Saak V
AU  - Ovsepian SV
FAU - Wang, Jiafu
AU  - Wang J
FAU - Zurawski, Tomas H
AU  - Zurawski TH
FAU - Schmidt, James J
AU  - Schmidt JJ
FAU - Smith, Leonard
AU  - Smith L
FAU - Lawrence, Gary W
AU  - Lawrence GW
FAU - Dolly, J Oliver
AU  - Dolly JO
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20120614
PL  - England
TA  - FEBS J
JT  - The FEBS journal
JID - 101229646
RN  - 0 (Liposomes)
RN  - 0 (Protease Inhibitors)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (SNARE Proteins)
RN  - 0 (Synaptosomal-Associated Protein 25)
RN  - 0Y70779M1F (rimabotulinumtoxinB)
RN  - 9013-20-1 (Streptavidin)
RN  - EC 3.4.24.69 (Botulinum Toxins)
RN  - EC 3.4.24.69 (Botulinum Toxins, Type A)
RN  - TPY09G7XIR (Fluorescein)
SB  - IM
MH  - Animals
MH  - Botulinum Toxins/chemistry/genetics/*metabolism
MH  - Botulinum Toxins, Type A
MH  - Diaphragm/metabolism
MH  - Drug Delivery Systems/*methods
MH  - Fluorescein/chemistry
MH  - Humans
MH  - Liposomes
MH  - Mice
MH  - Microscopy, Confocal
MH  - Motor Neurons/metabolism
MH  - Mutation
MH  - Nerve Endings/metabolism
MH  - Paralysis/metabolism/therapy
MH  - Presynaptic Terminals/*metabolism
MH  - Protease Inhibitors/*administration & dosage/chemistry
MH  - Recombinant Fusion Proteins/chemistry/genetics/metabolism
MH  - SNARE Proteins/*metabolism
MH  - Streptavidin/genetics/metabolism
MH  - Synaptosomal-Associated Protein 25/metabolism
EDAT- 2012/05/23 06:00
MHDA- 2012/10/09 06:00
CRDT- 2012/05/22 06:00
PHST- 2012/05/22 06:00 [entrez]
PHST- 2012/05/23 06:00 [pubmed]
PHST- 2012/10/09 06:00 [medline]
AID - 10.1111/j.1742-4658.2012.08638.x [doi]
PST - ppublish
SO  - FEBS J. 2012 Jul;279(14):2555-67. doi: 10.1111/j.1742-4658.2012.08638.x. Epub 
      2012 Jun 14.