Graduate NIH Training in Biotechnology
Former Trainees
Kyle G. Grant
PhD Chemical Engineering, University of Pittsburgh, April 2008.
Currently a Post-doc at the Medical Research Council, Virology Unit, Glasgow, Scotland.
Project Description
The work in our laboratory is based on the use of Herpes Simplex Virus as a gene therapy tool. In particular my project is centered on the use of complementing cell lines to propagate clinical grade virus in large quantities for use in upcoming clinical trials. These cell lines supply missing cytotoxic genes to our safe clinical vectors which allow the growth of replication defective vectors. HSV growth is regulated in an ordered cascade fashion by the production of certain viral proteins. Currently there is a 3-log order of magnitude reduction in viral production using the safe clinical vectors versus wild type virus. The bottleneck in production is due to a lack in efficient complementation of the missing toxic genes, namely ICP 4 and ICP 27, which are crucial for the cascade of regulation. Presently, effort is underway to model the viral infection in the complementing cell lines using RT-PCR, Western Blotting, and viral titers as readout of the transcriptional regulation, protein production, and viral production. This bottleneck has been somewhat ameliorated by the use of a more potent viral promoter from the ICP0 gene. Use of this promoter to drive the complementation of ICP4 has been shown to increase yields of more replication defective vectors by an order of magnitude. In addition, the use of different HDAC inhibitors has also been shown to increase yields by overcoming the repression mechanism of the cell to allow viral replication to ensue in the face of a strong anti-viral response. Efforts are under way to clone an inducible cell line encoding the ICP0 gene to overcome this repressive effect without the use of potentially toxic drugs.
Courses
Issues in Research and Teaching
Fundamentals of Thermodynamics
Fundamentals of Reaction Thermodynamics
Mathematical Methods in Chemical Engineering
Fundamentals of Transport Processes
Introduction to Cell and Molecular Biology (Biotechnology Training Program)
Scientific Ethics (Biotechnology Training Program)
Foundations of Biomedical Science (Biotechnology Training Program)
Foundations of Biomedical Science Conference (Biotechnology Training Program)
Molecular Virology
Herpes Virology
Biomaterials and Biocompatibility (Biotechnology Training Program)
Presentations
"Cell line development for production of clinical grade HSV vectors," American Chemical Society in San Diego, CA, March 2005, Biochemical Technology Division
”Advancement in cell line generation for production of replication defective HSV-1 vectors” American Institute of Chemical Engineers National Conference. Cincinnati, OH. October 2005 (Submitted and accepeted but withdrawn due to Industrial Internship).
“Cryopreservation of Primary Human Cell Lines” Cambrex Research and Development Department Update. Audience included Cambrex R&D from Walkersville, Rockland, and Nottingham UK. November 23rd 2005.
“Cell line engineering for the production of high replication defective HSV-1 vectors” American Society of Gene Therapy. Baltimore, MD. May 2006
“Engineering Cell Lines for the Production of Highly Replication Defective Herpes
Simplex Virus Type 1 Vectors” American Institute of Chemical Engineers.
San Francisco, CA November 2006
Seminar Attendance
Department of Chemical Engineering Graduate Seminar
McGowan Institute of Regenerative Medicine Wound Healing Seminar
McGowan Institute for Regenerative Medicine and Musculoskeletal
Research Center
Seminar Series
Advisor(s)
Mohammed Ataai
William Kepler Whiteford Professor, Department of Chemical Engineering and
Department of Bioengineering
University of Pittsburgh
Joseph C. Glorioso III
Chairman & William S. McEllroy Professor of Biochemistry
Department of Molecular Genetics and Biochemistry
University of Pittsburgh
