Gene Therapy and Vaccines Program

Address

Children's Hospital of Philadelphia
302 D Abramson Research Center
3615 Civic Center Boulevard
Philadelphia, PA 19104-4318

Office Tel: 215 590-4521
Fax: 215-590-3660
E-mail: high@email.chop.edu


Link(s)

The Cell and Gene Therapy Research Affinity Group Seminar Schedule (PDF)

Education


University of North Carolina School of Medicine, 1978. M.D.

Harvard College, 1972.A.B. (Chemistry)

University of Pennsylvania , 1993. MA (hon)

Research Interests

• 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.

Key words: Hemophilia B, AAV, Gene Therapy, Factor IX.

Search PubMed for articles

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.

Recent Publications

Margaritas P, Arruda VR, Aljamali, M, Camire R, Schlachterman A, High KA. Novel therapeutic approach for hemophilia using gene delivery of an engineered secreted activated FVII. J Clin Inv 113:1025-1031, 2004.

Sabatino D, Armstrong AE, Edmondson S, Pleimes M, Schuettrumpf, Fitzgerald J, Herzog RW, Arruda VR, High KA. Novel hemophilia B mouse models exhibiting a range of mutations in the Factor IX gene. Blood 104:2767-2774, 2004. [Epub 2004 Jun 24].

Arruda VR, Stedman HH, Nichols TC, Haskins ME, Nicholson M, Herzog RW, Couto LB, High KA. Regional intravascular infusion of AAV-2-F.IX to skeletal muscle achieves long-term correction of hemophilia B in a large animal model. Blood 105:3458-3464, 2005. [Epub ahead of print 2004 Oct. 12].

Sabatino DE, Mingozzi F, Hui DJ, Chen H, Colosi P, Ertl HCJ, High KA. Identification of mouse AAV capsid-specific CD8+ T cell epitopes. Mol Ther 12:1023-1033, 2005, [Epub ahead of print 2005 Nov. 2].

Manno CS, Arruda VR, Pierce GF, Glader B, Ragni M, Rasko J, Ozelo MC, Hoots K, Blatt, P, Konkle B, Dake M, Kaye R, Razavi M, Zajko A, Zehnder J, Nakai H, Chew A, Leonard D, Wright JF, Lessard RR, Sommer JM, Tigges M, Sabatino D, Luk A, Jiang H, Mingozzi F, Couto L, Ertl HC, High KA*, Kay MA. Successful transduction of liver in hemophilia by AAV-Factor IX and limitations imposed by the host immune response. Nat Med 12:342-347, 2006. *Corresponding author.

Lab

Rotation Projects for 2006-2007

1. Gene transfer for hemophilia B
2. Immunological aspects of viral vector-based gene transfer
3. Novel approaches to the treatment of hemophilia

Lab personnel:

Kirk Chu, Research Assoc
Shyrie Edmonson, Research Tech
Daniel Hui, Post Doc
Sonali Joyce, Research Tech
Jianhua Liu, Research Tech
Yi-Lin (Emily) Liu, Research Assoc
Paris Margaritis, Post Doc
Federico Mingozzi, Research Assoc
Samuel Murphy, Post Doc
Denise Sabatino, Research Associate
Alex Tai, Research Tech
Yi Zhao, Research Tech
Shang Zhen Zhou, Director, Research Vector Core