Single Tyrosine Mutation in AAV8 Vector Capsid Enhances Gene Lung Delivery and Does Not Alter Lung Morphofunction in Mice
Background/Aims: Vectors derived from adeno-associated viruses (AAVs) are important gene delivery tools for treating pulmonary diseases. Phosphorylation of surface-exposed tyrosine residues from AAV2 capsid targets the viral particles for ubiquitination and proteasome-mediated degradation, and mutat...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Cell Physiol Biochem Press GmbH & Co KG
2014-08-01
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| Series: | Cellular Physiology and Biochemistry |
| Subjects: | |
| Online Access: | http://www.karger.com/Article/FullText/363033 |
| _version_ | 1818507911988510720 |
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| author | Sabrina V. Martini Adriana L. da Silva Debora Ferreira Karina Gomes Felipe M. Ornellas Miquéias Lopes-Pacheco Emilia Zin Hilda Petrs-Silva Patricia R. M. Rocco Marcelo M. Morales |
| author_facet | Sabrina V. Martini Adriana L. da Silva Debora Ferreira Karina Gomes Felipe M. Ornellas Miquéias Lopes-Pacheco Emilia Zin Hilda Petrs-Silva Patricia R. M. Rocco Marcelo M. Morales |
| author_sort | Sabrina V. Martini |
| collection | DOAJ |
| description | Background/Aims: Vectors derived from adeno-associated viruses (AAVs) are important gene delivery tools for treating pulmonary diseases. Phosphorylation of surface-exposed tyrosine residues from AAV2 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. We evaluated the pulmonary transduction efficiency of AAV8 vectors containing point mutations in surface-exposed capsid tyrosine residues. Methods: Male C57BL/6 mice (20-25 g, n=24) were randomly assigned into three groups: control group animals received intratracheal (i.t.) instillation of saline (50 μl), wild-type AAV8 group, and capsid mutant Y733F AAV8 group, which received (i.t.) AAV8 vectors containing the DNA sequence of enhanced green fluorescence protein (eGFP). Four weeks after instillation, lung mechanics and morphometry, vector transduction (immunohistochemistry and mRNA expression of eGFP), and inflammatory cytokines and growth factor expression were analyzed. Results: Tyrosine-mutant AAV8 vectors displayed significantly increased transduction efficiency in the lung compared with their wild-type counterparts. No significant differences were observed in lung mechanics and morphometry between experimental groups. There was no evidence of inflammatory response in any group. Conclusion: AAV8 vectors may be useful for new therapeutic strategies for the treatment of pulmonary diseases. |
| first_indexed | 2024-12-10T22:24:39Z |
| format | Article |
| id | doaj.art-ccdddfa82e1e4861bb714835ced6be07 |
| institution | Directory Open Access Journal |
| issn | 1015-8987 1421-9778 |
| language | English |
| last_indexed | 2024-12-10T22:24:39Z |
| publishDate | 2014-08-01 |
| publisher | Cell Physiol Biochem Press GmbH & Co KG |
| record_format | Article |
| series | Cellular Physiology and Biochemistry |
| spelling | doaj.art-ccdddfa82e1e4861bb714835ced6be072022-12-22T01:31:13ZengCell Physiol Biochem Press GmbH & Co KGCellular Physiology and Biochemistry1015-89871421-97782014-08-0134368169010.1159/000363033363033Single Tyrosine Mutation in AAV8 Vector Capsid Enhances Gene Lung Delivery and Does Not Alter Lung Morphofunction in MiceSabrina V. MartiniAdriana L. da SilvaDebora FerreiraKarina GomesFelipe M. OrnellasMiquéias Lopes-PachecoEmilia ZinHilda Petrs-SilvaPatricia R. M. RoccoMarcelo M. MoralesBackground/Aims: Vectors derived from adeno-associated viruses (AAVs) are important gene delivery tools for treating pulmonary diseases. Phosphorylation of surface-exposed tyrosine residues from AAV2 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. We evaluated the pulmonary transduction efficiency of AAV8 vectors containing point mutations in surface-exposed capsid tyrosine residues. Methods: Male C57BL/6 mice (20-25 g, n=24) were randomly assigned into three groups: control group animals received intratracheal (i.t.) instillation of saline (50 μl), wild-type AAV8 group, and capsid mutant Y733F AAV8 group, which received (i.t.) AAV8 vectors containing the DNA sequence of enhanced green fluorescence protein (eGFP). Four weeks after instillation, lung mechanics and morphometry, vector transduction (immunohistochemistry and mRNA expression of eGFP), and inflammatory cytokines and growth factor expression were analyzed. Results: Tyrosine-mutant AAV8 vectors displayed significantly increased transduction efficiency in the lung compared with their wild-type counterparts. No significant differences were observed in lung mechanics and morphometry between experimental groups. There was no evidence of inflammatory response in any group. Conclusion: AAV8 vectors may be useful for new therapeutic strategies for the treatment of pulmonary diseases.http://www.karger.com/Article/FullText/363033Tyrosine mutantLung functionAdeno-associated virusGene therapy |
| spellingShingle | Sabrina V. Martini Adriana L. da Silva Debora Ferreira Karina Gomes Felipe M. Ornellas Miquéias Lopes-Pacheco Emilia Zin Hilda Petrs-Silva Patricia R. M. Rocco Marcelo M. Morales Single Tyrosine Mutation in AAV8 Vector Capsid Enhances Gene Lung Delivery and Does Not Alter Lung Morphofunction in Mice Cellular Physiology and Biochemistry Tyrosine mutant Lung function Adeno-associated virus Gene therapy |
| title | Single Tyrosine Mutation in AAV8 Vector Capsid Enhances Gene Lung Delivery and Does Not Alter Lung Morphofunction in Mice |
| title_full | Single Tyrosine Mutation in AAV8 Vector Capsid Enhances Gene Lung Delivery and Does Not Alter Lung Morphofunction in Mice |
| title_fullStr | Single Tyrosine Mutation in AAV8 Vector Capsid Enhances Gene Lung Delivery and Does Not Alter Lung Morphofunction in Mice |
| title_full_unstemmed | Single Tyrosine Mutation in AAV8 Vector Capsid Enhances Gene Lung Delivery and Does Not Alter Lung Morphofunction in Mice |
| title_short | Single Tyrosine Mutation in AAV8 Vector Capsid Enhances Gene Lung Delivery and Does Not Alter Lung Morphofunction in Mice |
| title_sort | single tyrosine mutation in aav8 vector capsid enhances gene lung delivery and does not alter lung morphofunction in mice |
| topic | Tyrosine mutant Lung function Adeno-associated virus Gene therapy |
| url | http://www.karger.com/Article/FullText/363033 |
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