Prostaglandin E2 Increases Lentiviral Vector Transduction Efficiency of Adult Human Hematopoietic Stem and Progenitor Cells

Abstract

Gene therapy currently in development for hemoglobinopathies utilizes ex vivo lentiviral transduction of CD34 hematopoietic stem and progenitor cells (HSPCs). A small-molecule screen identified prostaglandin E (PGE) as a positive mediator of lentiviral transduction of CD34 cells. Supplementation with PGE increased lentiviral vector (LVV) transduction of CD34 cells approximately 2-fold compared to control transduction methods with no effect on cell viability. Transduction efficiency was consistently increased in primary CD34 cells from multiple normal human donors and from patients with β-thalassemia or sickle cell disease. Notably, PGE increased transduction of repopulating human HSPCs in an immune-deficient (nonobese diabetic/severe combined immunodeficiency/interleukin-2 gamma receptor null [NSG]) xenotransplantation mouse model without evidence of in vivo toxicity, lineage bias, or a de novo bias of lentiviral integration sites. These data suggest that PGE improves lentiviral transduction and increases vector copy number, therefore resulting in increased transgene expression. As a result, PGE may be useful in clinical gene therapy applications using lentivirally modified HSPCs.

Genetic engineering of hematopoiesis: current stage of clinical translation and future perspectives

“…As compared to other cell types, HSC are poorly permissive to LV gene transfer, due to quiescence and the presence or enhanced expression of innate or adaptive restriction factors acting at different steps of the transduction pathway (Santoni de Sio et al, 2006;Wang et al, 2014). As more restriction factors are uncovered, new drugs become available to counteract them and enable enhanced (Wang et al, 2014;Petrillo et al, 2015;Zonari et al, 2017;Heffner et al, 2018) and better matched gene transfer among donors, as in the case of cyclosporin H, which overcomes IFITM3 restriction (Petrillo et al, 2018). Vector entry may occur from the plasma membrane or from endosomes following phagocytosis, as dictated by the choice of envelope pseudotype, and is subject to differential restriction.…”

“…More recently, IFN-inducible antiviral factors, such as IFITM3, have been shown to impair LV entry into HSPC according to donor-dependent expression level (Petrillo et al, 2018). As more restriction factors are uncovered, new drugs become available to counteract them and enable enhanced (Wang et al, 2014;Petrillo et al, 2015;Zonari et al, 2017;Heffner et al, 2018) and better matched gene transfer among donors, as in the case of cyclosporin H, which overcomes IFITM3 restriction (Petrillo et al, 2018).…”

Mesenchymal stromal cell‐derived extracellular vesicles as cell‐free biologics for the ex vivo expansion of hematopoietic stem cells

“…To overcome this limitation, various studies have been performed using different approaches, such as the use of a combination of cytokines (Lui et al, 2014), growth factors (Buza‐Vidas et al, 2006; Hofmeister et al, 2007; Himburg et al, 2010), virus‐mediated expression of certain genes, metabolic modulation, MSC‐HSC co‐cultures, genetic modification or inducing a hypoxic or oxidative environment, etc. (Goessling et al, 2011; Farahbakhshian et al, 2014; Xiao et al, 2015; Kumar and Geiger, 2017; Heffner et al, 2018).…”

Transplantation of gene-modified haematopoietic stem cells: Application and clinical considerations

“…Optimisation of lentiviral transduction protocols over the years has resulted in significant improvements in transduction rates, including the use of high MOIs (>100), transduction over 2 consecutive days, optimised growth media and cytokine usage, [18][19][20] and transduction enhancers. [21,22] …”