Comparative intra-articular gene transfer of seven adeno-associated virus serotypes reveals that AAV2 mediates the most efficient transduction to mouse arthritic chondrocytes

Quan Chen; Huan Luo; Chengcong Zhou; Huan Yu; Sai Yao; Fangda Fu; Rebecca Seeley; Xing Ji; Yanping Yang; Peifeng Chen; Hongting Jin; Peijian Tong; Di Chen; Chengliang Wu; Weibin Du; Hongfeng Ruan

doi:10.1371/journal.pone.0243359

Comparative intra-articular gene transfer of seven adeno-associated virus serotypes reveals that AAV2 mediates the most efficient transduction to mouse arthritic chondrocytes

PLoS One. 2020 Dec 15;15(12):e0243359. doi: 10.1371/journal.pone.0243359. eCollection 2020.

Authors

Quan Chen^{1

2}, Huan Luo³, Chengcong Zhou^{1

2}, Huan Yu^{1

2}, Sai Yao^{1

2}, Fangda Fu^{1

2}, Rebecca Seeley⁴, Xing Ji⁴, Yanping Yang⁵, Peifeng Chen^{1

2}, Hongting Jin^{1

2}, Peijian Tong^{1

2}, Di Chen⁶, Chengliang Wu^{1

2}, Weibin Du⁷, Hongfeng Ruan^{1

2

5}

Affiliations

¹ The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, China.
² The First Clinical Medical College of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, China.
³ Department of Pharmacy, The Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
⁴ Translational Research Program in Pediatric Orthopedics, The Children's Hospital of Philadelphia, Philadelphia, PA, United States of America.
⁵ Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, China.
⁶ Research Center for Human Tissues and Organs Degeneration, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.
⁷ Research Institute of Orthopedics, the Affiliated JiangNan Hospital of Zhejiang Chinese Medical University, Hangzhou, China.

Abstract

Osteoarthritis (OA) is the most common arthropathy, characterized by progressive degeneration of the articular cartilage. Currently, there are no disease-modifying approaches for OA treatment. Adeno-associated virus (AAV)-mediated gene therapy has recently become a potential treatment for OA due to its exceptional characteristics; however, the tropism and transduction efficiency of different AAV serotypes to articular joints and the safety profile of AAV applications are still unknown. The present study aims to screen an ideal AAV serotype to efficiently transfer genes to arthritic cartilage. AAV vectors of different serotypes expressing eGFP protein were injected into the knee joint cavities of mice, with all joint tissues collected 30 days after AAV injection. The transduction efficiency of AAVs was quantified by assessing the fluorescent intensities of eGFP in the cartilage of knee joints. Structural and morphological changes were analyzed by toluidine blue staining. Changes to ECM metabolism and pyroptosis of chondrocytes were determined by immunohistochemical staining. Fluorescence analysis of eGFP showed that eGFP was expressed in the cartilage of knee joints injected with each AAV vector. Quantification of eGFP intensity indicated that AAV2, 7 and 8 had the highest transduction efficiencies. Both toluidine blue staining and Mankin score showed that AAV6 aggravated cartilage degeneration. The analysis of key molecules in ECM metabolism suggested that AAV5 and 7 significantly reduced collagen type II, while AAV9 increased ADAMTS-4 but decreased MMP-19. In addition, transduction with AAV2, 5, 7 and 8 had no obvious effect on pyroptosis of chondrocytes. Comprehensive score analysis also showed that AAV2 had the highest score in intra-articular gene transfer. Collectively, our findings point to AAV2 as the best AAV serotype candidate for gene transfer on arthritic cartilage, resulting in minimal impact to ECM metabolism and pyroptosis of chondrocytes.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

ADAMTS4 Protein / biosynthesis
ADAMTS4 Protein / genetics
Animals
Arthritis, Experimental* / genetics
Arthritis, Experimental* / metabolism
Arthritis, Experimental* / pathology
Arthritis, Experimental* / therapy
Cartilage, Articular / metabolism*
Cartilage, Articular / pathology
Chondrocytes / metabolism*
Chondrocytes / pathology
Collagen Type II / biosynthesis
Collagen Type II / genetics
Dependovirus
Genetic Therapy*
Genetic Vectors*
Knee Joint / metabolism*
Knee Joint / pathology
Male
Matrix Metalloproteinases, Secreted / biosynthesis
Matrix Metalloproteinases, Secreted / genetics
Mice
Parvovirinae*
Transduction, Genetic

Substances

Collagen Type II
Matrix Metalloproteinases, Secreted
matrix metalloproteinase 19
ADAMTS4 Protein
Adamts4 protein, mouse

Supplementary concepts

Adeno-associated virus-2

Grants and funding

This study was financially supported by the following grants: National Natural Science Foundation of China (No.: 81804121 to HR, 81973870 and 81573994 to CW, 81904053 to WD, 81973877 and 81674006 to YY); China Postdoctoral Science Foundation (No.: 2018M632154 to HR); Natural Science Foundation of Zhejiang Province (No.: LY19H270006 to CW, LY19H290004 to MY, and LQ17H270005 to HR); Traditional Chinese Medical Administration of Zhejiang Province (No.: 2021ZB090 and 2017ZB026 to HR); Zhejiang Medical and Health Science and Technology Project (No.: 2018269058 to HL and 2021ZB090 to HR); Zhejiang Chinese Medical University Scientific Research Fund Project (No.: 2018ZG20 to CW); Postgraduate Science Research Fund of Zhejiang Chinese Medical University (2020YKJ07 to CZ); National Undergraduate Innovation and Entrepreneurship Training Program (202010344004 to FF and SY); Opening Project of Zhejiang Provincial Preponderant and Characteristic Subject of Key University (Chinese Traditional Medicine) and Zhejiang Chinese Medical University (No.: ZYX2018008 to WD); Plan Guide Project of HangZhou Technology Department (No.: 20171226Y98 to WD); Plan technology project of Hangzhou Health Department (No.: 2018B028 to WD); and Science and Technology Innovation Program for College Students (New talent program) of Zhejiang Province (No.: 2018R410030 to HR. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.