Gene transfer of a therapeutic protein is a promising approach toward treatment of genetic disease. Adenoassociated? viral (AAV) mediated in vivo transfer of a coagulation factor IX (F.IX) gene to muscle or liver of? subjects with severe hemophilia B is currently being evaluated in clinical trials. A major concern of treatment? is induction of CDS* cytotoxic T lymphocyte responses (CTL), which may cause elimination of transduced? cells (muscle fibers or hepatocytes) expressing the transgene product. Interestingly, AAV gene transfer has? been shown not to cause CTL responses to a number of transgene products, which may be due to immune? tolerance or ignorance. We will determine requirements for tolerance induction to a human F.IX transgene? product in mice, which will receive AAV-F.IX gene transfer followed by challenge with an adenoviral vector (a? potent vector for induction of CTL) expressing the identical transgene. CTL responses will be measured by? antigen-specific lysis of target cells and immunochemical analyses of transduced tissues. We will define? requirements for tolerance induction (target organ, levels of transgene expression, systemic protein? delivery). We hypothesize that AAV-mediated hepatic gene transfer resulting in systemic F.IX protein? delivery can lead to immune tolerance thereby preventing CTL responses to F.IX. Furthermore, we? hypothesize that endogenous expression of immunodominant F.IX T cell epitopes contributes to tolerance? thereby reducing the risk of CTL responses. The effect of the underlying mutation in endogenously? expressed F.IX on CTL responses will be tested in a series of transgenic mice expressing non-functional? variants of human F.IX taken from the hemophilia B database. Finally, a series of experiments is; proposed to? identify subsets of regulatory T cells that can suppress development of CTL responses to F.IX. These? studies are based on the hypothesis that hepatic AAV-F.IX gene transfer induces regulatory CD4+ T cells? capable of suppressing CTL to F.IX. A detailed characterization of these regulatory cells and the? requirements for their induction will lead to improved gene transfer protocols and will be an experimental? basis for approaches to augment regulatory T cell responses. Improved understanding of cellular? mechanisms underlying immune responses or absence thereof should provide the basis for design of? strategies that preclude destruction of cells receiving viral gene transfer.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Program Projects (P01)
Project #
Application #
Study Section
Special Emphasis Panel (ZHL1)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Wistar Institute
United States
Zip Code
High, Katherine A; Anguela, Xavier M (2016) Adeno-associated viral vectors for the treatment of hemophilia. Hum Mol Genet 25:R36-41
Vercauteren, Koen; Hoffman, Brad E; Zolotukhin, Irene et al. (2016) Superior In vivo Transduction of Human Hepatocytes Using Engineered AAV3 Capsid. Mol Ther 24:1042-1049
Wang, Xiaomei; Terhorst, Cox; Herzog, Roland W (2016) In vivo induction of regulatory T cells for immune tolerance in hemophilia. Cell Immunol 301:18-29
Rogers, Geoffrey L; Suzuki, Masataka; Zolotukhin, Irene et al. (2015) Unique Roles of TLR9- and MyD88-Dependent and -Independent Pathways in Adaptive Immune Responses to AAV-Mediated Gene Transfer. J Innate Immun 7:302-14
Rogers, Geoffrey L; Herzog, Roland W (2015) Gene therapy for hemophilia. Front Biosci (Landmark Ed) 20:556-603
Hui, Daniel J; Edmonson, Shyrie C; Podsakoff, Gregory M et al. (2015) AAV capsid CD8+ T-cell epitopes are highly conserved across AAV serotypes. Mol Ther Methods Clin Dev 2:15029
Wang, Xiaomei; Su, Jin; Sherman, Alexandra et al. (2015) Plant-based oral tolerance to hemophilia therapy employs a complex immune regulatory response including LAP+CD4+ T cells. Blood 125:2418-27
Sharma, Rajiv; Anguela, Xavier M; Doyon, Yannick et al. (2015) In vivo genome editing of the albumin locus as a platform for protein replacement therapy. Blood 126:1777-84
Biswas, Moanaro; Terhorst, Cox; Herzog, Roland W (2015) Treg: tolerance vs immunity. Oncotarget 6:19956-7
Biswas, Moanaro; Sarkar, Debalina; Kumar, Sandeep R P et al. (2015) Synergy between rapamycin and FLT3 ligand enhances plasmacytoid dendritic cell-dependent induction of CD4+CD25+FoxP3+ Treg. Blood 125:2937-47

Showing the most recent 10 out of 74 publications