Original Article

Gene Therapy (2017) 24, 290–297; doi:10.1038/gt.2017.19; published online 13 April 2017

Optimization of adeno-associated virus vector-mediated gene transfer to the respiratory tract

F Kurosaki1,2, R Uchibori1,3, N Mato2, Y Sehara1, Y Saga1,4, M Urabe1, H Mizukami1, Y Sugiyama2 and A Kume5

  1. 1Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Japan
  2. 2Division of Pulmonary Medicine, Department of Medicine, Jichi Medical University, Shimotsuke, Japan
  3. 3Division of Immuno-Gene and Cell Therapy (Takara Bio), Jichi Medical University, Shimotsuke, Japan
  4. 4Department of Obstetrics and Gynecology, Jichi Medical University, Shimotsuke, Japan
  5. 5Support Center for Clinical Investigation, Jichi Medical University, Shimotsuke, Japan

Correspondence: Professor A Kume, Support Center for Clinical Investigation, Jichi Medical University, 3311-1 Yakushiji, Shimotsuke 329-0498, Japan. E-mail: kume@jichi.ac.jp

Received 31 August 2016; Revised 16 March 2017; Accepted 17 March 2017
Accepted article preview online 27 March 2017; Advance online publication 13 April 2017



An efficient adeno-associated virus (AAV) vector was constructed for the treatment of respiratory diseases. AAV serotypes, promoters and routes of administration potentially influencing the efficiency of gene transfer to airway cells were examined in the present study. Among the nine AAV serotypes (AAV1–9) screened in vitro and four serotypes (AAV1, 2, 6, 9) evaluated in vivo, AAV6 showed the strongest transgene expression. As for promoters, the cytomegalovirus (CMV) early enhancer/chicken β-actin (CAG) promoter resulted in more robust transduction than the CMV promoter. Regarding delivery routes, intratracheal administration resulted in strong transgene expression in the lung, whereas the intravenous and intranasal administration routes yielded negligible expression. The combination of the AAV6 capsid and CAG promoter resulted in sustained expression, and the intratracheally administered AAV6-CAG vector transduced bronchial cells and pericytes in the lung. These results suggest that AAV6-CAG vectors are more promising than the previously preferred AAV2 vectors for airway transduction, particularly when administered into the trachea. The present study offers an optimized strategy for AAV-mediated gene therapy for lung diseases, such as cystic fibrosis and pulmonary fibrosis.