Ultrasound-Mediated Gene Therapy for Hemophilia B
Principal Investigator: CAROL MIAO
Abstract: This proposal aims at developing ultrasound-mediated nonviral gene delivery to the liver for treatment of hemophilia and other genetic diseases. We have shown that ultrasound (US) in the presence of microbubbles [MBs] enhances gene delivery of a liver-specific, high-expressing factor IX (FIX) plasmid into mice. Significant enhancement was achieved by transducing one liver lobe, a 66-fold increment relative to use of naked DNA alone. Mouse livers could also be transduced with a GFP plasmid. Therapeutic US together with MBs therefore has the potential to promote safe and efficient nonviral gene transfer of hFIX for the treatment of hemophilia. To facilitate translation of this method to human applications, we propose to further optimize the US parameters for higher efficacy. The optimized protocols, together with optimized pDNA/MBs complexes, improved FIX gene transfer vectors, will be tested in a hemophilia B mouse model to achieve a long-term therapeutic effect. Suitable US systems will be designed for testing the US protocols in normal rats, normal and hemophilia B dogs. We will test the hypotheses that: 1) By combining the best US protocol and optimized FIX gene expression cassette, therapeutic levels of FIX expression can be achieved in hemophilia B mice and can correct their phenotype long-term. 2) US parameters and probes in larger animal models can be optimized to achieve high-level reporter gene expression in normal dog models. 3) By combining the best US protocol and optimized cFIX plasmids, phenotypic correction of hemophilia B dogs can be achieved. 4) Focused US or shockwave therapy will be suitable for translation of the US protocol into clinical application. Public Health Relevance: Our goal is to develop an Ultrasound-mediated gene therapy protocol that can be easily translated into human applications to treat hemophilia and other genetic diseases. We have previously shown that delivery of a high-expressing, liver-specific hFIX plasmid can be enhanced by ultrasound (US) in combination with microbubbles (MBs) and achieved 66 fold increment in expression levels compared with gene transfer without US and MBs. This proposal will continue optimizing our US protocol in mouse and rat models and extend the development to normal and hemophilia B dog models.
Funding Period: 2009-06-08 - 2010-08-31
more information: NIH RePORT
- Immunomodulation for inhibitors in hemophilia A: the important role of Treg cellsCarol H Miao
Seattle Children s Research Institute and Department of Pediatrics, University of Washington, Seattle, WA, USA
Expert Rev Hematol 3:469-83. 2010..Innovative strategies to overcome pre-existing anti-FVIII immune responses and induce long-term tolerance in primed subjects still need to be developed...
- Efficient microbubble- and ultrasound-mediated plasmid DNA delivery into a specific rat liver lobe via a targeted injection and acoustic exposure using a novel ultrasound systemShuxian Song
Seattle Children s Research Institute, Seattle, Washington
Mol Pharm 9:2187-96. 2012..These results indicated that unfocused high intensity therapeutic ultrasound exposure with microbubbles is highly promising for safe and efficient gene delivery into the liver of rats or larger animals. ..
- Ultrasound-targeted microbubble destruction-mediated gene delivery into canine liversMisty L Noble
Center for Immunity and Immunotherapies, Seattle Children s Research Institute, Seattle, Washington 98101, USA
Mol Ther 21:1687-94. 2013..7 MPa. Transaminase and histology analysis indicated minimal tissue damage. These experiments represent an important developmental step toward US-mediated gene delivery in large animals and clinics...
- Development of therapeutic microbubbles for enhancing ultrasound-mediated gene deliveryRyan R Sun
Center for Immunity and Immunotherapies, Seattle Children s Research Institute, Seattle, USA
J Control Release 182:111-20. 2014..With further designs, MB customizations have the potential to advance this technology closer to clinical application. ..
- Explorations of high-intensity therapeutic ultrasound and microbubble-mediated gene delivery in mouse liverS Song
Department of Pediatrics, Seattle Children s Research Institute, University of Washington, Seattle, WA 98101, USA
Gene Ther 18:1006-14. 2011..This study validated the new HITU system equipped with an unfocused transducer with a larger footprint capable of scanning large tissue areas to effectively enhance gene transfer efficiencies...