Abstract
Background aims
Gene-modified mesenchymal stromal cells (MSC) provide a promising tool for cell and
gene therapy-based applications by potentially acting as a cellular vehicle for protein-replacement
therapy. However, to avoid the risk of insertional mutagenesis, targeted integration
of a transgene into a ‘safe harbor’ locus is of great interest.
Methods
We sought to determine whether zinc finger nuclease (ZFN)-mediated targeted addition
of the erythropoietin (Epo) gene into the chemokine [C-C motif] receptor 5 (CCR5)
gene locus, a putative safe harbor locus, in MSC would result in stable transgene
expression in vivo.
Results
Whether derived from bone marrow (BM), umbilical cord blood (UCB) or adipose tissue
(AT), 30–40% of human MSC underwent ZFN-driven targeted gene addition, as determined
by a combination of fluorescence-activated cell sorting (FACS)- and polymerase chain
reaction (PCR)-based analyzes. An enzyme-linked immunosorbent assay (ELISA)-based
analysis of gene-targeted MSC expressing Epo from the CCR5 locus showed that these
modified MSC were found to secrete a significant level of Epo (c. 2 IU/106cells/24 h). NOD/SCID/γC mice injected with ZFN-modified MSC expressing Epo exhibited
significantly higher hematocrit and Epo plasma levels for several weeks post-injection,
compared with mice receiving control MSC.
Conclusions
These data demonstrate that MSC modified by ZFN-driven targeted gene addition may
represent a cellular vehicle for delivery of plasma-soluble therapeutic factors.
Key Words
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References
- Adipose-tissue engineering: taking advantage of the properties of human adipose-derived stem/stromal cells.Pathol Biol (Paris). 2009; 57: 309-317
- Isolation and characterization of rapidly self-renewing stem cells from cultures of human marrow stromal cells.Cytotherapy. 2001; 3: 393-396
- Derivation of lung epithelium from human cord blood-derived mesenchymal stem cells.Am J Respir Crit Care Med. 2008; 177: 701-711
- IFATS collection: using human adipose-derived stem/stromal cells for the production of new skin substitutes.Stem Cells. 2008; 26: 2713-2723
- Multilineage cells from human adipose tissue: implications for cell-based therapies.Tissue Eng. 2001; 7: 211-228
- Plasticity in adipogenesis and osteogenesis of human mesenchymal stem cells.Mol Cell Endocrinol. 2007; 271: 1-17
- Mesenchymal stem cells: properties and role in clinical bone marrow transplantation.Curr Opin Immunol. 2006; 18: 586-591
- Human-compatible collagen matrix for prolonged and reversible systemic delivery of erythropoietin in mice from gene-modified marrow stromal cells.Mol Ther. 2004; 10: 741-748
- Generation of insulin-producing cells from human bone marrow mesenchymal stem cells by genetic manipulation.Stem Cells. 2007; 25: 2837-2844
- LMO2-associated clonal T cell proliferation in two patients after gene therapy for SCID-X1.Science. 2003; 302: 415-419
- Insertional mutagenesis combined with acquired somatic mutations causes leukemogenesis following gene therapy of SCID-X1 patients.J Clin Invest. 2008; 118: 3143-3150
- Gene editing in human stem cells using zinc finger nucleases and integrase-defective lentiviral vector delivery.Nat Biotechnol. 2007; 25: 1298-1306
- Targeted gene addition into a specified location in the human genome using designed zinc finger nucleases.Proc Natl Acad Sci USA. 2007; 104: 3055-3060
- Highly efficient endogenous human gene correction using designed zinc-finger nucleases.Nature. 2005; 435: 646-651
- Establishment of HIV-1 resistance in CD4+ T cells by genome editing using zinc-finger nucleases.Nat Biotechnol. 2008; 26: 808-816
- Mammalian gene targeting with designed zinc finger nucleases.Mol Ther. 2006; 13: 438-446
- Resistance to HIV-1 infection in caucasian individuals bearing mutant alleles of the CCR-5 chemokine receptor gene.Nature. 1996; 382: 722-725
- Effective gene therapy with nonintegrating lentiviral vectors.Nat Med. 2006; 12: 348-353
- In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector.Science. 1996; 272: 263-267
- Correction of multi-gene deficiency in vivo using a single ‘self-cleaving’ 2A peptide-based retroviral vector.Nat Biotechnol. 2004; 22: 589-594
- Mesenchymal stromal cells engineered to express erythropoietin induce anti-erythropoietin antibodies and anemia in allorecipients.Mol Ther. 2009; 17: 369-372
Article info
Publication history
Accepted:
December 23,
2009
Received:
September 10,
2009
Identification
Copyright
© 2010 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.
