This application seeks funding for studies aimed at dissecting the down-regulation of adaptive immune responses directed against coagulation factors that are introduced by protein or gene replacement therapy into individuals with the X-linked bleeding disorder hemophilia. These potentially detrimental responses not only result in the formation of inhibitory antibodies against factor VIII (deficient in hemophilia A) or factor IX (F.IX, deficient in hemophilia B), they also include the generation of cytotoxic T lymphocytes, which can eliminate cells that are modified upon gene transfer. Whilst both sets of responses are dependent on antigen specific helper CD4+ T cells, adaptive immune responses to F.IX are suppressed by naive or antigen specific regulatory T cells (Treg, Tr1, NKT). Nonetheless, we are only beginning to understand the implications of immune regulation in the treatment of hemophilia, as we need to dissect the genetic and mechanistic underpinnings of these regulatory processes. The experiments proposed in this Competing Renewal application are designed to test the general hypothesis that several regulatory T cell subsets cooperate in the prevention of a primary response or mitigate an existing response to F.IX introduced via AAV-based gene transfer or as a protein. The proposed experiments are grouped into the following Specific Aims: i) To optimize the in vitro generation of F.IX specific Treg. ii) To test the hypothesis that in vivo antibody and CD8+ T cell responses directed against F.IX can be suppressed by induction by Treg. iii) To test the hypothesis that conventional Treg, IL-10 producing Treg and NKT cells regulate immune responses directed against AAV-hF.IX. Together these experiments should clarify the basic mechanisms, which govern inhibitor and T cell responses to coagulation factors and therapeutic proteins. The results could directly be used for the design of therapeutic strategies that can be applied treatment of hemophilia.

Public Health Relevance

The immune system can respond to hemophilia therapies and therefore also represents a hurdle for novel gene therapies. This proposal aims to establish novel ways to achieve tolerance to the therapeutic clotting factors by taking advantage of immune cells that actively suppress immune responses.

National Institute of Health (NIH)
Research Program Projects (P01)
Project #
Application #
Study Section
Heart, Lung, and Blood Program Project Review Committee (HLBP)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Wistar Institute
United States
Zip Code
Rogers, Geoffrey L; Martino, Ashley T; Zolotukhin, Irene et al. (2014) Role of the vector genome and underlying factor IX mutation in immune responses to AAV gene therapy for hemophilia B. J Transl Med 12:25
Sarkar, Debalina; Biswas, Moanaro; Liao, Gongxian et al. (2014) Ex Vivo Expanded Autologous Polyclonal Regulatory T Cells Suppress Inhibitor Formation in Hemophilia. Mol Ther Methods Clin Dev 1:
Wang, Xiaomei; Moghimi, Babak; Zolotukhin, Irene et al. (2014) Immune tolerance induction to factor IX through B cell gene transfer: TLR9 signaling delineates between tolerogenic and immunogenic B cells. Mol Ther 22:1139-50
Liao, Gongxian; van Driel, Boaz; Magelky, Erica et al. (2014) Glucocorticoid-induced TNF receptor family-related protein ligand regulates the migration of monocytes to the inflamed intestine. FASEB J 28:474-84
Liao, Gongxian; O'Keeffe, Michael S; Wang, Guoxing et al. (2014) Glucocorticoid-Induced TNF Receptor Family-Related Protein Ligand is Requisite for Optimal Functioning of Regulatory CD4(+) T Cells. Front Immunol 5:35
Wu, Te-Lang; Li, Hua; Faust, Susan M et al. (2014) CD8+ T cell recognition of epitopes within the capsid of adeno-associated virus 8-based gene transfer vectors depends on vectors' genome. Mol Ther 22:42-51
Sherman, Alexandra; Schlachterman, Alexander; Cooper, Mario et al. (2014) Portal vein delivery of viral vectors for gene therapy for hemophilia. Methods Mol Biol 1114:413-26
Mingozzi, Federico; Anguela, Xavier M; Pavani, Giulia et al. (2013) Overcoming preexisting humoral immunity to AAV using capsid decoys. Sci Transl Med 5:194ra92
Buchlis, George; Odorizzi, Pamela; Soto, Paula C et al. (2013) Enhanced T cell function in a mouse model of human glycosylation. J Immunol 191:228-37
Mingozzi, Federico; High, Katherine A (2013) Immune responses to AAV vectors: overcoming barriers to successful gene therapy. Blood 122:23-36

Showing the most recent 10 out of 51 publications