Primary Research Area: Stem Cell Biology/Hematopoiesis
The research focus in our laboratory is to formulate innovative gene and cell therapeutic approaches for the treatment of hemophiliacs. There are three major areas that we are working on.
Gene therapy of hemophilia A
Hemophilia A is a recessive X-linked bleeding disorder resulting from a FVIII deficiency. Although protein replacement therapy is effective for hemophilia A treatment, up to 35% of patients may develop inhibitory antibodies (referred to as inhibitors) that neutralize FVIII activity. Introducing FVIII expression via genetic therapy is an attractive alternative treatment for hemophilia A patients. However, the potential to develop inhibitors to the transgene protein remains a significant concern. We have recently developed a novel gene therapy approach in which FVIII is targeted and stored in platelet α-granules. Using a transgenic mouse model, we showed that FVIII can be specifically expressed and stored together with its carrier protein, VWF, in platelet α-granules when it is driven by the platelet-specific αIIb promoter (2bF8) and that platelet-FVIII can correct the murine hemophilia A phenotype even in the presence of high-titer inhibitors. To apply this gene therapy to a clinically translatable protocol, we use lentiviral gene delivery to hematopoietic stem cells (HSCs) to introduce FVIII expression in platelets. We found that 2bF8 lentiviral gene delivery to HSCs can not only restore hemostasis, but also induce antigen-specific immune tolerance in hemophilia A mice. Our current work aims to 1) further optimize this approach; 2) dissect the potential underlying mechanisms by which immune tolerance is induced after platelet gene therapy; and 3) translate findings made in laboratory animals to human patients.
Gene therapy of hemophilia B
Hemophilia B is a genetic bleeding disorder which results from a FIX deficiency. Although the incidence of anti-FIX inhibitor development is lower (5%) in hemophilia B patients after protein replacement therapy, anaphylaxis is a daunting problem in inhibitor patients, which limits the use of FIX infusion and increases the risk of morbidity and mortality. Our studies show that FIX can be ectopically expressed and stored in platelet α-granules when it is driven under the same αIIb promoter that we use for platelet-FVIII expression. Platelet-derived FIX, which is fully γ-carboxylated, can rescue bleeding diathesis in hemophilia B mice, but the clinical efficacy is limited in the presence of anti-FIX inhibitors. Although platelet-FIX does not maintain clinical efficacy in the face of inhibitors, targeting FIX expression to platelets is still an attractive potential strategy for gene therapy of hemophilia B. Indeed, our studies demonstrate that 2bF9 gene delivery to HSCs not only restores hemostasis, but also induce FIX-specific immune tolerance in hemophilia B mice. Our current work aims to optimize the FIX expression and develop a protocol for gene therapy of hemophilia B with pre-existing immunity.
VWF/FVIII association and FVIII immunity
While the important association between VWF and FVIII has been investigated for decades, how VWF impacts FVIII immunity is still uncertain. Our studies demonstrate that VWF is essential for maintaining platelet-FVIII efficacy in hemophilia A with inhibitors. VWF reduces inhibitor inactivation of FVIII, exerting a protective effect both in vitro and in vivo. VWF/FVIII association plus the apparent ability of VWF to delay the time-dependent inactivation of FVIII by inhibitors provides mechanisms by which platelet-derived FVIII maintains function even in the presence of inhibitors when FVIII is targeted to platelets for gene therapy of hemophilia A with inhibitors. Our current work aims to investigate the roles of VWF in FVIII immunity in hemophilia A.