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Preparation and Gene Modification of Nonhuman Primate Hematopoietic Stem and Progenitor Cells
Published on: February 15, 2019
Engineered human Tmpk fused with truncated cell-surface markers: versatile cell-fate control safety cassettes
M Scaife1, N Pacienza, B C Y Au
1Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.
Gene Therapy
|January 14, 2012
Summary
This study introduces a novel fusion system for cell-fate control gene therapy (CFCGT), linking cell marking and eradication functions. This approach enhances safety in gene and cell therapies by ensuring a direct correlation between marking and elimination.
Area of Science:
- Biotechnology
- Gene Therapy
- Molecular Biology
Background:
- Existing gene therapies and cell transplantation methods can benefit from enhanced safety features.
- Previous work established a novel enzyme/prodrug combination for cell-fate control gene therapy (CFCGT).
Purpose of the Study:
- To develop and validate a novel lentiviral construct for CFCGT that enforces a one-to-one correlation between cell marking and eradication.
- To assess the in vitro and in vivo efficacy and safety of this fusion system in various models, including a proof-of-principle study for Fabry disease.
Main Methods:
- Constructed novel lentiviral vectors fusing truncated surface molecules (CD19 or low-affinity nerve growth factor receptor) to the CFCGT cDNA (TmpkF105Y).
- Performed in vitro functional analysis of the fusion product.
- Evaluated in vivo efficacy in NOD/SCID mice with K562 cells and a murine leukemia-lymphoma model with Raji cells, using 3'-azido-3'-deoxythymidine (AZT) for prodrug administration.
- Integrated the CFCGT fusion construct into a lentivector for Fabry disease treatment and tested it on Fabry cells and patient-derived CD34(+) cells.
Main Results:
- In vitro studies confirmed the functionality of the fusion product.
- In vivo studies in mice demonstrated tumor growth suppression and cytotoxicity in leukemia-lymphoma models.
- The lentivector for Fabry disease restored enzyme activity in affected cells and maintained prodrug sensitivity.
- High transduction efficiencies and normal colony-forming capacity were observed in human Fabry patient CD34(+) cells.
Conclusions:
- The novel fusion system establishes a direct link between cell marking and eradication, enhancing safety in gene and cell therapies.
- This broadly applicable system shows promise for improving the safety profile of various gene and cell-based therapeutic strategies.
- The technology is validated in preclinical models and shows potential for treating genetic disorders like Fabry disease.
