PMID- 27384068
OWN - NLM
STAT- MEDLINE
DCOM- 20170210
LR  - 20190212
IS  - 1421-9778 (Electronic)
IS  - 1015-8987 (Linking)
VI  - 39
IP  - 2
DP  - 2016
TI  - Tyrosine Mutation in AAV9 Capsid Improves Gene Transfer to the Mouse Lung.
PG  - 544-53
LID - 10.1159/000445646 [doi]
AB  - BACKGROUND/AIMS: Adeno-associated virus (AAV) vectors are being increasingly used 
      as the vector of choice for in vivo gene delivery and gene therapy for many 
      pulmonary diseases. Recently, it was shown that phosphorylation of 
      surface-exposed tyrosine residues from AAV capsid targets the viral particles for 
      ubiquitination and proteasome-mediated degradation, and mutations of these 
      tyrosine residues lead to highly efficient vector transduction in vitro and in 
      vivo in different organs. In this study, we evaluated the pulmonary transgene 
      expression efficacy of AAV9 vectors containing point mutations in surface-exposed 
      capsid tyrosine residues. METHODS: Eighteen C57BL/6 mice were randomly assigned 
      into three groups: (1) a control group (CTRL) animals underwent intratracheal 
      (i.t.) instillation of saline, (2) the wild-type AAV9 group (WT-AAV9, 1010 vg), 
      and (3) the tyrosine-mutant Y731F AAV9 group (M-AAV9, 1010 vg), which received 
      (i.t.) self-complementary AAV9 vectors containing the DNA sequence of enhanced 
      green fluorescence protein (eGFP). Four weeks after instillation, lung mechanics, 
      morphometry, tissue cellularity, gene expression, inflammatory cytokines, and 
      growth factor expression were analyzed. RESULTS: No significant differences were 
      observed in lung mechanics and morphometry among the experimental groups. 
      However, the number of polymorphonuclear cells was higher in the WT-AAV9 group 
      than in the CTRL and M-AAV9 groups, suggesting that the administration of 
      tyrosine-mutant AAV9 vectors was better tolerated. Tyrosine-mutant AAV9 vectors 
      significantly improved transgene delivery to the lung (30%) compared with their 
      wild-type counterparts, without eliciting an inflammatory response. CONCLUSION: 
      Our results provide the impetus for further studies to exploit the use of AAV9 
      vectors as a tool for pulmonary gene therapy.
CI  - (c) 2016 The Author(s) Published by S. Karger AG, Basel.
FAU - Martini, Sabrina V
AU  - Martini SV
AD  - Laboratory of Cellular and Molecular Physiology, Carlos Chagas Filho Institute of 
      Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, Ilha do 
      Fundx00E3;o, Brazil.
FAU - Silva, Adriana L
AU  - Silva AL
FAU - Ferreira, Debora
AU  - Ferreira D
FAU - Rabelo, Rafael
AU  - Rabelo R
FAU - Ornellas, Felipe M
AU  - Ornellas FM
FAU - Gomes, Karina
AU  - Gomes K
FAU - Rocco, Patricia R M
AU  - Rocco PR
FAU - Petrs-Silva, Hilda
AU  - Petrs-Silva H
FAU - Morales, Marcelo M
AU  - Morales MM
LA  - eng
PT  - Journal Article
DEP - 20160707
PL  - Germany
TA  - Cell Physiol Biochem
JT  - Cellular physiology and biochemistry : international journal of experimental 
      cellular physiology, biochemistry, and pharmacology
JID - 9113221
RN  - 0 (Capsid Proteins)
RN  - 0 (Cytokines)
RN  - 0 (Intercellular Signaling Peptides and Proteins)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - 42HK56048U (Tyrosine)
SB  - IM
MH  - Animals
MH  - Capsid Proteins/*genetics
MH  - Cytokines/genetics/metabolism
MH  - Dependovirus/*genetics
MH  - Gene Expression
MH  - Genetic Therapy/methods
MH  - Genetic Vectors/genetics
MH  - Green Fluorescent Proteins/genetics/metabolism
MH  - Intercellular Signaling Peptides and Proteins/genetics/metabolism
MH  - Lung/*metabolism
MH  - Male
MH  - Mice, Inbred C57BL
MH  - *Point Mutation
MH  - Random Allocation
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Transfection/*methods
MH  - Transgenes/genetics
MH  - Tyrosine/*genetics
EDAT- 2016/07/08 06:00
MHDA- 2017/02/12 06:00
CRDT- 2016/07/08 06:00
PHST- 2016/05/31 00:00 [accepted]
PHST- 2016/07/08 06:00 [entrez]
PHST- 2016/07/08 06:00 [pubmed]
PHST- 2017/02/12 06:00 [medline]
AID - 000445646 [pii]
AID - 10.1159/000445646 [doi]
PST - ppublish
SO  - Cell Physiol Biochem. 2016;39(2):544-53. doi: 10.1159/000445646. Epub 2016 Jul 7.