Adeno-associated virus (AAV) vector has been successfully applied in phase I clinical trials in hemophilia B patients with liver targeting. However, these studies have suggested that AAV capsid specific cytotoxic T lymphocytes (CTL) have the potential to eliminate AAV transduced hepatocytes and result in the therapeutic failure. Our prior studies have demonstrated that AAV capsid antigen presentation is dose-dependent and requires capsid ubiquitination for proteasome mediated degradation. The contamination of empty virions in AAV preparation inhibits transduction from full particles of AAV vectors and potentially increases the risk of virus capsid antigen load. In this proposal we will investigate capsid antige presentation from AAV empty virions and the effect of empty particles on antigen presentation from full virus transduction (Aim 1). To decrease antigen presentation on AAV transduced cells for avoiding capsid specific CTL-mediated elimination, it has been proposed to modify the AAV capsid surface or apply proteasome inhibitors to enhance AAV transduction while lowering the effective dose or to escape capsid ubiquitination. We will study the effect of AAV mutants and proteasome inhibitors on AAV capsid antigen presentation (Aim 2). It is well-known that the transduction of AAV vectors in mouse models does not always translate into the human. Finally, we will explore the directed evolution approach combined with a rational design strategy to isolate AAV vectors with human hepatocyte specific tropism and the ability to evade a capsid specific CTL response in humanized mice (Aim 3). Elucidation of AAV empty capsid antigen presentation in vivo and the development of an AAV vector with enhanced human liver transduction and CTL immune-evasion will allow us to design safer and more effective strategies that address the current clinical complications for human liver gene therapy using AAV.

Public Health Relevance

Having demonstrated that AAV capsid antigen presentation is dose-dependent and requires proteasome mediated degradation, and modification of the AAV capsid surface induces enhanced AAV transduction while lowering the effective dose or decreases capsid antigen presentation, we will explore to develop AAV mutants with the ability to evade capsid specific CTL mediated elimination and with human hepatocyte tropism. This study will allow us to design safer and more effective strategies for human liver gene therapy using AAV.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI117408-04
Application #
9655276
Study Section
Gene and Drug Delivery Systems Study Section (GDD)
Program Officer
Gondre-Lewis, Timothy A
Project Start
2016-03-01
Project End
2021-02-28
Budget Start
2019-03-01
Budget End
2020-02-29
Support Year
4
Fiscal Year
2019
Total Cost
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Pediatrics
Type
Schools of Medicine
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Zhang, Xintao; He, Ting; Chai, Zheng et al. (2018) Blood-brain barrier shuttle peptides enhance AAV transduction in the brain after systemic administration. Biomaterials 176:71-83
Sun, Junjiang; Shao, Wenwei; Chen, Xiaojing et al. (2018) An Observational Study from Long-Term AAV Re-administration in Two Hemophilia Dogs. Mol Ther Methods Clin Dev 10:257-267
Chai, Zheng; Zhang, Xintao; Rigsbee, Kelly Michelle et al. (2018) Cryoprecipitate augments the global transduction of the adeno-associated virus serotype 9 after a systemic administration. J Control Release 286:415-424
Pei, Xiaolei; He, Ting; Hall, Nikita E et al. (2018) AAV8 virions hijack serum proteins to increase hepatocyte binding for transduction enhancement. Virology 518:95-102
Shao, Wenwei; Chen, Xiaojing; Samulski, Richard J et al. (2018) Inhibition of antigen presentation during AAV gene therapy using virus peptides. Hum Mol Genet 27:601-613
Shao, Wenwei; Earley, Lauriel F; Chai, Zheng et al. (2018) Double-stranded RNA innate immune response activation from long-term adeno-associated virus vector transduction. JCI Insight 3:
Sun, Junjiang; Hua, Baolai; Chen, Xiaojing et al. (2017) Gene Delivery of Activated Factor VII Using Alternative Adeno-Associated Virus Serotype Improves Hemostasis in Hemophiliac Mice with FVIII Inhibitors and Adeno-Associated Virus Neutralizing Antibodies. Hum Gene Ther 28:654-666
Sanchez, Anamaria C; Li, Chengwen; Andrews, Barbara et al. (2017) AAV Gene Therapy for Alcoholism: Inhibition of Mitochondrial Aldehyde Dehydrogenase Enzyme Expression in Hepatoma Cells. Hum Gene Ther 28:717-725
Liang, Katharine J; Woodard, Kenton T; Weaver, Mark A et al. (2017) AAV-Nrf2 Promotes Protection and Recovery in Animal Models of Oxidative Stress. Mol Ther 25:765-779
Wang, M; Sun, J; Crosby, A et al. (2017) Direct interaction of human serum proteins with AAV virions to enhance AAV transduction: immediate impact on clinical applications. Gene Ther 24:49-59

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