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Member Information
Katherine A. High, MD
William H. Bennett Professor of Pediatrics
Children's Hospital of Philadelphia
Office Phone: 215-590-4521
Office Fax:
Email: high@email.chop.edu
Website(s):
Education: MD 1978, University of North Carolina - Chapel Hill
Keywords: Hemophilia, AAV, HIV, HCV
Research and/or Clinical Interests:
Molecular basis of blood coagulation; gene transfer as an approach to treating hemophilia; influence of antecedent HIV and HCV infection on viral-vector mediated gene transfer for hematologic disorders.
Research Interest Summary:
The focus of the laboratory is on the genetics, molecular biology and biochemistry of blood coagulation factors. We have concentrated primarily on the vitamin K dependent factors, Factor VII, IX and X. 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.
Representative Publications:
Herzog RW, Arruda VR, Fisher TH, Read MS, Nichols TC, and High KA. Absence of circulating factor IX antigen in hemophilia B dogs of the UNC-Chapel Hill colony. Thrombosis and Haemostasis 84: 352-354, 2000.
Camire, R.J., Larson PJ,, Stafford DW, High KA. Enhanced ?-carboxylation of recombinant Factor X using a chimeric construct containing the prothrombin propeptide. Biochemistry , 39: 14322-14329, 2000.
Arruda VR, Hagstrom JN, Herzog RW, Heiman-Patterson T, Deitch J, Chu K, Couto LB, Larson PJ, Camire R and High KA Post-translational modifications of recombinant myotube-synthesized human factor IX. Blood 97:130-138, 2001.
Hung, H.L. Pollak ES, Kudaravalli RD, Chu K and High K.A. Regulation of human coagulation factor X gene expression by GATA-4 and the Sp family of transcription factors. Blood , 97: 946-951, 2001.
Fields P, Armstrong EA, Hagstrom JN, Arruda VR, Murphy ML, Farrell JP, High KA, Herzog RW. Intravenous administration of an E1,E3-deleted adenoviral vector induces tolerance to a Factor IX transgene in C57BL/6 mice. Gene Therapy , 8: 354-361, 2001.
Fields PA, Arruda VR, Armstrong E, Chu K, Mingozzi F, Hagstrom JN, Herzog RW, High, KA. Risk and prevention of anti-factor IX formation in AAV-mediated gene transfer in context of a large factor IX gene deletion. Molecular Therapy , 4: 201-210, 2001.
Arruda, VR, Fields PA, Milner R, Wainwright L, deMiguel MP, Donovan PJ, Herzog RW, Nichols TC, Biegel J, Razavi M, Huff D, Flake AW, Couto L, Kay MA, High KA. Lack of germline transmission of vector sequences following systemic administration of recombinant AAV-2 vector in males. Molecular Therapy , 4: 586-592, 2001.
Mount MJ, Herzog RW, Tillson M, Goodman SA, Robinson N, McCleland ML, Bellinger D, Nichols TC, Arruda VR, Lothrop Jr. CD, High KA. Sustained high-level correction of inhibitor-prone hemophilia B dogs by liver-directed gene therapy. Blood 99: 2670-2676, 2002. See commentary Blood 99:2635.
Herzog RW, Fields PA, Arruda VR, Brewbaker J, Armstrong EA, McClintock D, Bellinger D, Couto LB, Nichols TC, High KA. Influence of vector dose on Factor IX-specific T and B cell responses in muscle-directed gene therapy for hemophilia B. Human Gene Therapy Vol. 13: 1281-1291, 2002.
Toso R, Bernardi F, Tidd T, Pinotti M, Camire RM, Marchetti G, High KA, Pollak ES. Factor VII mutant V154G models a zymogen-like form of Factor VIIa. Biochem J 369: 563-571, 2003.
Manno CS, Chew A, Hutchison S, Larson PJ, Herzog RW, Arruda VR, Tai SJ, Ragni MV, Thompson A, Ozelo M, Couto LB, Leonard DGB, Johnson FA, McClelland A, Scallan C, Skarsgaard, E, Flake AW, Kay MA, High KA, Glader B. AAV-mediated factor IX gene transfer to skeletal muscle in patients with severe hemophilia B. Blood 2002 Dec. 19 (epub ahead of print), 101: 2963-2972, 2003.
High KA, Herzog R, Arruda V. AAV-mediated gene transfer to liver. Blood 101: 3338-3339, 2003.
Li B, Lawler AM, Antonarakis SE, High KA, Gearhart JD and Kazazian HH: Targeted disruption of the mouse factor VIII gene: A model for hemophilia A. Nature Genetics 10: 119-121, 1995.
Pollak E, Hung HL, Godin W, Overton GC and High KA: Functional characterization of the human Factor VII 5' flanking region. J Biol Chem 271:1738-1747, 1996.
Hung HL and High KA: Liver-enriched transcription factor HNF-4 and ubiquitous factor NF-Y are critical for expression of blood coagulation factor X. J Biol Chem 271:2323-2330, 1996.
Larson P, Stanfield-Oakley, S, VanDusen WA , Kasper C, Smith K, Monroe , DM, High KA: Structural integrity of the gla domain of human blood coagulation factor IXa is required for its binding to cofactor factor VIIIa. J Biol Chem 271:3869-3875, 1996.
Walter J, You Q, Hagstrom N, Sands M, High KA: Successful expression of human Factor IX following repeat administration of an adenoviral vector in mice. PNAS 93:3056-3061, 1996.
