Tyrosine-Mutant AAV8 Vector Mediated Efficient and Safe Gene Transfer of Pigment Epithelium-Derived Factor to Mouse Lungs

Débora P Ferreira; Sabrina V Martini; Helena A Oliveira; Adriana L Silva; Siddharth Shenoy; Daiqin Chen; Valentina Simon; Eric Han; Natalie E West; Jung Soo Suk; Patricia R M Rocco; Hilda Petrs-Silva; Marcelo M Morales; Fernanda F Cruz

doi:10.33594/000000660

Tyrosine-Mutant AAV8 Vector Mediated Efficient and Safe Gene Transfer of Pigment Epithelium-Derived Factor to Mouse Lungs

Cell Physiol Biochem. 2023 Sep 18;57(5):331-344. doi: 10.33594/000000660.

Authors

Débora P Ferreira^{1

2

3}, Sabrina V Martini¹, Helena A Oliveira⁴, Adriana L Silva^{1

2

3}, Siddharth Shenoy^{5

6}, Daiqin Chen^{5

6}, Valentina Simon⁵, Eric Han^{5

6}, Natalie E West⁷, Jung Soo Suk^{5

6

8}, Patricia R M Rocco^{2

3}, Hilda Petrs-Silva⁹, Marcelo M Morales^{1

3}, Fernanda F Cruz^{10

3}

Affiliations

¹ Laboratory of Cellular and Molecular Physiology, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
² Laboratory of Pulmonary Investigation, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
³ National Institute of Science and Technology for Regenerative Medicine, Rio de Janeiro, RJ, Brazil.
⁴ Laboratory of Immunopharmacology, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro, Brazil.
⁵ Center for Nanomedicine, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
⁶ Department of Ophthalmology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
⁷ Department of Medicine, Johns Hopkins University, Baltimore, MD, USA.
⁸ Department of Chemical and Biomolecular Engineering, Whiting School of Engineering, Johns Hopkins University, Baltimore, MD, USA.
⁹ Laboratory of Gene Therapy and Viral Vectors, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
¹⁰ Laboratory of Pulmonary Investigation, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil, ffcruz@biof.ufrj.br.

PMID: 37724045
DOI: 10.33594/000000660

Abstract

Background/aims: Recombinant adeno-associated viruses (rAAV) are an important tool for lung targeted gene therapy. Substitution of tyrosine with phenylalanine residues (Y-F) in the capsid have been shown to protect the AAV vector from ubiquitin/proteasome degradation, increasing transduction efficiency. We tested the mutant Y733F-AAV8 vector for mucus diffusion, as well as the safety and efficacy of pigment epithelium-derived factor (PEDF) gene transfer to the lung.

Methods: For this purpose, Y733F-AAV8-PEDF (10¹⁰ viral genome) was administered intratracheally to C57BL/6 mice. Lung mechanics, morphometry, and inflammation were evaluated 7, 14, 21, and 28 days after injection.

Results: The tyrosine-mutant AAV8 vector was efficient at penetrating mucus in ex vivo assays and at transferring the gene to lung cells after in vivo instillation. Increased levels of transgene mRNA were observed 28 days after vector administration. Overexpression of PEDF did not affect in vivo lung parameters.

Conclusion: These findings provide a basis for further development of Y733F-AAV8-based gene therapies for safe and effective delivery of PEDF, which has anti-angiogenic, anti-inflammatory and anti-fibrotic activities and might be a promising therapy for lung inflammatory disorders.

Keywords: Gene therapy; Recombinant adeno-associated viruses serotype 8 (AAV8); Tyrosine mutant; Pigment epithelium-derived factor (PEDF).

MeSH terms

Animals
Eye Proteins* / genetics
Gene Transfer Techniques*
Mice
Mice, Inbred C57BL
Nerve Growth Factors / genetics
Serpins* / genetics

Substances

Eye Proteins
Nerve Growth Factors
pigment epithelium-derived factor
Serpins

Abstract

MeSH terms

Substances

Grants and funding