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Enhanced transduction of Macaca fascicularis hematopoietic cells with chimeric lentiviral vectors.

Human Gene Therapy 2019 March 9
Recent marketing approval for genetically engineered hematopoietic stem and T cells bears witness to the substantial improvements in lentiviral vectors over the last two decades, but evaluations of the long-term efficacy and toxicity of gene and cell therapy products will, nevertheless, require further studies in nonhuman primate models. <i>Macaca fascicularis</i> monkeys from Mauritius have a low genetic diversity and are particularly useful for reproducible drug testing. In particular, they have a genetically homogeneous class I MHC system that probably mitigates the variability of the response to simian immunodeficiency virus infection. However, the transduction of simian cells with human immunodeficiency virus 1-derived vectors is inefficient, due to capsid-specific restriction factors, such as the tripartite motif-containing protein TRIM5α, which prevent infection with non-host-adapted retroviruses. We introduced the modified capsid of the macaque-trophic HIV-1 clone MN4/LSQD into our packaging system and compared transduction efficiencies between hematopoietic cells transduced with this construct and cells transduced with HIV-1 NL4-3-derived packaging constructs. Capsid modification increased transduction efficiency in all hematopoietic cells tested (by factors of up to 10), including hematopoietic progenitor cells, repopulating cells, and T cells from Mauritian <i>Macaca fascicularis</i>, regardless of vector structure or purification method. We also established culture conditions similar to those used in clinical practice for the efficient transduction of hematopoietic stem and progenitor CD34<sup>+</sup> cells. These results suggest that the procedure is suitable for use in Mauritian <i>Macaca fascicularis</i>, which can, therefore, be used as a model in preclinical studies for hematopoietic gene and cell therapy.

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