Hematology, hemostasis and thrombosis, gene transfer for genetic disease
Dr. High, a world renowned hematologist, studies the molecular basis of blood coagulation and the development of novel therapeutics for the treatment of bleeding disorders. Her pioneering bench-to-bedside studies of gene therapy for hemophilia have led to a focus on clinical translation of genetic therapies for hemophilia and other rare, inherited disorders. As the Director of the Center for Cellular and Molecular Therapeutics at The Children?s Hospital of Philadelphia, Dr. High leads a multidisciplinary team of scientists and researchers working to discover new gene and cell therapies for genetic diseases and to facilitate rapid translation of pre-clinical discoveries into clinical application.
The focus of the laboratory was initially on the molecular basis of blood coagulation, and the use of novel genetic therapies to treat hemophilia. More recently we have pioneered safe and effective clinical translation of genetic therapies for inherited disorders. These clinical trials have led to short-term correction of disease in hemophilia B, and long-term improvements in Leber?s congenital amaurosis, a hereditary cause of blindness.
Key words: Hemophilia , AAV, Gene Therapy, Factor IX, Factor VIIa.
Description of Research
Major areas of investigation include: 1) structure-function analysis of Factors VII, IX and X through the study of naturally occurring mutant proteins and recombinant proteins produced using site-directed mutagenesis and a mammalian expression system, 2) study of the regulation of expression of the genes encoding the vitamin K dependent clotting factors. These genes manifest tissue-specific expression; they are expressed only in the liver. We have isolated the 5' flanking sequences of all three genes, characterized promoter activity and determined, using DNase footprinting, the location of protein binding sites within these promoters. In the case of Factor X, we have determined through a variety of approaches the identity of the transcription factors binding to the promoter and are in the process of carrying out similar studies on the promoters of VII and IX, 3) studies designed to establish an experimental and clinical basis for gene therapy of hemophilia, a bleeding disorder that results from a deficiency of functional Factor IX. We are currently using both viral vectors to introduce the Factor IX cDNA into target cells of interest. My colleagues and I have several active projects, including a study on the generation and use of recombinant AAV vectors expressing Factor IX to treat hemophilia B. In particular, we are investigating novel methods of delivering vector to target tissues, and are also exploring the use of alternate serotypes and optimized expression cassettes in order to maximize gene expression. Other work focuses on safety problems, including determinants of the immune response to the transgene product, and assessment of risk of germline transmission of vector sequences. In other experiments, we are investigating the function of coagulation proteins through the use of targeted disruption of clotting factor genes. Current areas of interest include completion of clinical trials of an AAV-mediated, liver directed approach.
Rotation Projects for 2013-2014
1. Gene transfer for hemophilia B
2. Immunological aspects of viral vector-based gene transfer
3. Novel approaches to the treatment of hemophilia
Xavier Anguela, Post Doc;
Etiena Basner-Tschakarjan, Research Assoc;
George Buchlis, Post Doc;
Yifeng Chen, Research Assoc;
Robert Davidson, Research Assoc;
Liron Elkouby, Research Assoc;
Daniel Hui, Post Doc;
Jianhua Liu, Research Tech;
Rajiv Sharma, Pre Doc;
Alex Tai, Research Tech;
Yi Zhao, Research Tech;
Shangzhen Zhou, Director, Research Vector Core
- William H. Bennett Associate Professor of Pediatrics at University of Pennsylvania School of Medicine (1996 – 1999)
- Associate Professor of Pathology and Laboratory Medicine at University of Pennsylvania School of Medicine (1993 – 1998)
- Associate Professor of Medicine at University of Pennsylvania School of Medicine (1996 – 2001)
- William H. Bennett Professor of Pediatrics at University of Pennsylvania School of Medicine (1999 – 2006)
- Professor of Pediatrics at University of Pennsylvania School of Medicine (2006 – 2009)
- Associate Professor of Pediatrics at University of Pennsylvania School of Medicine (1993 – 1996)
- William H. Bennett Professor of Pediatrics at University of Pennsylvania School of Medicine (2009– present)
- Professor of Medicine in Medicine at University of Pennsylvania School of Medicine (2001 – 2012)
- MA (hon), University of Pennsylvania (1993)
- Cert, Business, UNC Business School. Management Institute for Hospital Administrators. (1989)
- M.D., Medicine, University of North Carolina School of Medicine (1978)
- A.B., Chemistry, Harvard College (1972)
- Mingozzi F, Anguela XM, Pavani G, Chen Y, Davidson RJ, Hui DJ, Yazicioglu M, Elkouby L, Hinderer CJ, Faella A, Howard C, Tai A, Podsakoff GM, Zhou S, Basner-Tschakarjan E, Wright JF, High KA. Overcoming preexisting humoral immunity to AAV using capsid decoys.. Sci Transl Med. Vol 5(194) . 2013 Jul:194ra92.
- Yazicioglu MN, Monaldini L, Chu K, Khazi F, Murphy SL, Huang H, Margaritis P, High, KA. Cellular localization and characterization of cytosolic binding partners for Gla-containing proteins PRRG4 and PRRG2.. J Biol Chem. 2013 Jul 19 [Epub ahead of print].
- Gil-Farina I, DiScala M, Vanrell L, Olagüe, Vales A, High KA, Prieto J, Mingozzi J, Gonzelez-Aseguinolaza G.. IL12-Mediated liver inflammation reduces the formation of AAV transcriptionally active forms but has no effect over preexisting AAV transgene expression.. PLoS One. Vol 8(7) . 2013 Jul:e67748.
- Buchlis G, Odorizzi P, Soto PC, Pearce OM, Hui DJ, Jordan MS, Varki A, Wherry EJ, High KA. Enhanced T cell function in a mouse model of human glycosylation. J Immunol. Vol 191(1) . 2013 Jul:228-237.
- Mingozzi F, Chen Y, Edmonson SC, Zhou S, Thurlings RM, Tak PP, High KA. Prevalence and pharmacological modulation of humoral immunity to AAV vectors in gene transfer to synovial tissue. Gene Ther. Vol 20. 2013 Jul:417-424.
- Testa F, Maguire AM, Rossi S, Pierce EA, Melillo P, Marshall K, Banfi S, Surace EM, Sun J, Acerra C, Wright JF, Wellman J, High KA, Auricchio A, Bennett J, Simonelli F.. Three-year follow-up after unilateral subretinal delivery of adeno-associated virus in patients with Leber Congenital Amaurosis Type 2. Ophthalmology. Vol 120(6) . 2013 June:1283-1291.
- Hoffman BA, Ertl HCJ, Terhorst C, High KA, Herzog RW. Gene therapy at the frontiers of viral immunology. Front Microbiol. Vol 3. 2013 May:182.
- Callejas D, Mann CJ, Ayuso E, Lage R, Grifoll I, Roca C, Andaluz A, Ruiz-de Gopegui R, Montane J, Munoz S, Ferre T, Haurigot V, Zhou S, Ruberte J, Mingozzi F, High K, Garcia F, Bosch F. Treatment of diabetes and long-term survival following insulin and glucokinase gene therapy. Diabetes. Vol 62(5) . 2013 May:1718-1729.
- Mingozzi F. and High KA.. Immune responses for AAV vectors: overcoming barriers to successful gene therapy. Blood. Vol 122(1) . 2013 Apr:23-36.
- Parzych EM, Liu H, Yin X, Liu Q, Wu TL, Podsakoff G, High KA, Levine MH, Ertl HC. Effects of immunosupression on circulating AAV capsid-specific T cells in humans. Hum Gene Ther. Vol 24. 2013 Apr:431-422.