229. PGE2 Increases Lentiviral Vector Transduction Efficiency of Human HSC

Heffner, Garrett C.; Bonner, Melissa; Campbell, Dakota; Christiansen, Lauryn; Pierciey, Francis J.; Zhang, Wen; Lewis, Gretchen; Smurnyy, Yegor; Hamel, Amanda; Shah, Syed Imran Ali; Horton, Holly M.; Ryu, Byoung Y.; Goss, Kendrick A.; Nègre, Olivier; Veres, Gábor; Horvath, Christopher; Finer, Mitchell; Gregory, Philip D.

doi:10.1016/s1525-0016(16)33038-6

Cited by 2 publications

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“…Applying PGE 2 makes shorter ex vivo manipulation protocols that feature a single transduction cycle clinically viable, mitigating the negative effect of prolonged culture on progenitor cell engraftment and allowing to double the number of patients that can be treated with a given vector batch with respect to current clinical standards. Importantly, the enhancing effect of PGE 2 on LV transduction has recently been reported by an independent group ( Heffner et al., 2016 ). In contrast to other pharmacological interventions that enhance transduction but may also have broader effects on HSC biology and/or cause toxicity ( Wang et al., 2014 , Petrillo et al., 2015 ), PGE 2 was non-toxic, did not change the biological properties of the cells, and has already been safely employed in an ex vivo setting to increase CB engraftment in patients ( Cutler et al., 2013 ).…”

Section: Discussionmentioning

confidence: 56%

Efficient Ex Vivo Engineering and Expansion of Highly Purified Human Hematopoietic Stem and Progenitor Cell Populations for Gene Therapy

et al. 2017

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SummaryEx vivo gene therapy based on CD34+ hematopoietic stem cells (HSCs) has shown promising results in clinical trials, but genetic engineering to high levels and in large scale remains challenging. We devised a sorting strategy that captures more than 90% of HSC activity in less than 10% of mobilized peripheral blood (mPB) CD34+ cells, and modeled a transplantation protocol based on highly purified, genetically engineered HSCs co-infused with uncultured progenitor cells. Prostaglandin E2 stimulation allowed near-complete transduction of HSCs with lentiviral vectors during a culture time of less than 38 hr, mitigating the negative impact of standard culture on progenitor cell function. Exploiting the pyrimidoindole derivative UM171, we show that transduced mPB CD34+CD38− cells with repopulating potential could be expanded ex vivo. Implementing these findings in clinical gene therapy protocols will improve the efficacy, safety, and sustainability of gene therapy and generate new opportunities in the field of gene editing.

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Section: Discussionmentioning

confidence: 56%

Efficient Ex Vivo Engineering and Expansion of Highly Purified Human Hematopoietic Stem and Progenitor Cell Populations for Gene Therapy

et al. 2017

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“…The cell transduction efficiency of LVs is difficult to control and predict, even with purified vectors, and is seemingly highly dependent on patient samples 8, 54, 55. Finding agents to enhance the frequency of hematopoietic cell transduction is being considered,56, 57 but higher transduction efficacy on the fraction of cells highly permissive to transduction may considerably increase the VCN per transduced cells and the risk for insertional mutagenesis 58 . Heterozygous β 0 /β + -thalassemia carriers are healthy.…”

Section: Discussionmentioning

confidence: 99%

Ex Vivo Selection of Transduced Hematopoietic Stem Cells for Gene Therapy of β-Hemoglobinopathies

et al. 2018

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Although gene transfer to hematopoietic stem cells (HSCs) has shown therapeutic efficacy in recent trials for several individuals with inherited disorders, transduction incompleteness of the HSC population remains a hurdle to yield a cure for all patients with reasonably low integrated vector numbers. In previous attempts at HSC selection, massive loss of transduced HSCs, contamination with non-transduced cells, or lack of applicability to large cell populations has rendered the procedures out of reach for human applications. Here, we fused codon-optimized puromycin N-acetyltransferase to herpes simplex virus thymidine kinase. When expressed from a ubiquitous promoter within a complex lentiviral vector comprising the β-globin gene, viral titers and therapeutic gene expression were maintained at effective levels. Complete selection and preservation of transduced HSCs were achieved after brief exposure to puromycin in the presence of MDR1 blocking agents, suggesting the procedure's suitability for human clinical applications while affording the additional safety of conditional suicide.

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229. PGE2 Increases Lentiviral Vector Transduction Efficiency of Human HSC

Cited by 2 publications

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Efficient Ex Vivo Engineering and Expansion of Highly Purified Human Hematopoietic Stem and Progenitor Cell Populations for Gene Therapy

Efficient Ex Vivo Engineering and Expansion of Highly Purified Human Hematopoietic Stem and Progenitor Cell Populations for Gene Therapy

Ex Vivo Selection of Transduced Hematopoietic Stem Cells for Gene Therapy of β-Hemoglobinopathies

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