Functional aspects of factor VIII expression after transplantation of genetically-modified hematopoietic stem cells for hemophilia A

Lucienne M. Ide, Neal N. Iwakoshi, Bagirath Gangadharan, Shawn Jobe, Robert Moot, David McCarty, Christopher B. Doering, H. Trent Spencer

Research output: Contribution to journalArticlepeer-review

32 Scopus citations


Background: Major complications with respect to the development of gene therapy treatments for hemophilia A include low factor VIII (fVIII) expression and humoral immune responses resulting in inhibitory anti-fVIII antibodies. We previously achieved sustained curative fVIII activity levels in hemophilia A mice after nonmyeloablative transplantation of genetically-modified hematopoietic stem cells (HSCs) encoding a B-domain deleted porcine fVIII (BDDpfVIII) transgene with no evidence of an immune response. Methods: Mouse HSCs were transduced using MSCV-based recombinant virus encoding BDDpfVIII and transplanted into hemophilia A mice. Transplanted mice were followed for donor cell engraftment, fVIII expression and activity, and generation of anti-fVIII immune response. Results: We now show that: (i) the protein expressed by hematopoietic cells has a specific activity similar to that of purified protein; (ii) BDDpfVIII expressed from hematopoietic cells effectively induces thrombus formation, which is shown using a new method of in vivo analysis of fVIII function; (iii) naïve and pre-immunizedmice receiving HSC gene therapy are nonresponsive to challenges with recombinant human fVIII; (iv) nonresponsiveness is not broken after stringent challenges with BDDpfVIII; and (v) T cells from these mice are unresponsive to BDDpfVIII presentation. Furthermore, stem cells isolated from donors with high titer anti-human fVIII antibodies show no defects in donor cell engraftment or the ability to express BDDpfVIII. Conclusions: These results demonstrate that HSC gene therapy can be an effective alternative treatment for individuals with hemophilia A and may benefit patients by inducing immunological nonresponsiveness to fVIII replacement products.

Original languageEnglish (US)
Pages (from-to)333-344
Number of pages12
JournalJournal of Gene Medicine
Issue number4
StatePublished - Apr 2010
Externally publishedYes


  • Factor VIII
  • Gene therapy
  • Hematopoietic stem cells
  • Hemophilia A
  • Recombinant retrovirus


Dive into the research topics of 'Functional aspects of factor VIII expression after transplantation of genetically-modified hematopoietic stem cells for hemophilia A'. Together they form a unique fingerprint.

Cite this