CLINICAL PROTOCOL 
Gene Therapy for Gaucher Disease: Ex Vivo Gene Transfer 
and Autologous Transplantation of CD34 + Cells 
Principal Investigator: John A. Barranger, M.D., Ph.D. 
Co-Investigators: Edward Ball, M.D. TO OCO 
Sallie Boggs, Ph.D. 
Paul D. Robbins, Ph.D. 
Alfred Bahnson, Ph.D. 
Maya Nimgaonkar, M.D. 
Erin 0. Rice, M.S. 
Institution: University of Pittsburgh 
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TABLE OF CONTENTS 
Page 
I. Lay Summary ii 
II. Scientific Summary iii 
III. Background 
A. Gaucher Disease 1 
B. G1 ucocerebrosidase 1 
C. Gene Transfer/Gene Therapy 2 
IV. Pre-Clinical Studies 
A. Definition of the Target Cell and Efficacy of 
the Gene Product 3 
B. Vector Constructions 3 
C. Ex Vivo Gene Transfer and Syngeneic Bone Marrow 
Transplantation in Mice 5 
D. Transduction and Expression of the Human GC 
Gene in Normal and Gaucher Patient Macrophages 8 
E. Gene Transfer and Expression in CD34 + Cells from Blood 9 
V. Experimental Design 
A. Overview 11 
B. Patient Selection 12 
C. Description and Characterization of the Vector 
for Gene Transfer 13 
D. Production and Characterization of the 
Viral Producer Line 13 
E. Production and Characterization of the Viral Supernatant ... 15 
F. G-CSF Priming and Collection of a Mononuclear Cell Fraction. 16 
G. Enrichment of CD34 + Cells 16 
H. Transduction of CD34 + Cells 16 
I. Measurement of Transduction Efficiency and Expression 17 
J. Testing of Final Product for Bacterial and Viral 
Contamination 17 
K. Infusion of Transduced Cells 17 
L. Estimation of Engraftment 18 
M. Response Criteria 19 
N. Assessment of Risk/Benefit 20 
VI. References 22 
VII. Figures 25 
VIII. Responses to Points to Consider PCI 
IX. IRB Submission IRB1 
X. Appendix A1 
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Recombinant DNA Research, Volume 17 
