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

Garrett C Heffner,Yegor Smurnyy,Dakota Campbell,Seema Shaw,Gabor Veres,Melissa Bonner,Philip D Gregory,Lauryn Christiansen,Holly Horton,Amanda Hamel,Mitchell H Finer,Kendrick A Goss,Francis J Pierciey,Wenliang Zhang,Olivia Garijo,Gretchen Lewis,Bruce E Torbett

doi:10.1016/j.ymthe.2017.09.025

Garrett C Heffner, Yegor Smurnyy + Show 15 more

Open Access

https://doi.org/10.1016/j.ymthe.2017.09.025

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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 E2 (PGE2) as a positive mediator of lentiviral transduction of CD34+ cells. Supplementation with PGE2 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, PGE2 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 PGE2 improves lentiviral transduction and increases vector copy number, therefore resulting in increased transgene expression. As a result, PGE2 may be useful in clinical gene therapy applications using lentivirally modified HSPCs.

Highlights

Hematopoietic stem cell transplantation is a potentially curative therapy for multiple clinical indications
To facilitate the potential for rapid implementation in a Good Manufacturing Practice process, we selected the ScreenWell US Food and Drug Administration (FDA)-approved Drug Library v2 (Enzo Life Sciences), which contained more than 780 compounds, including known antiretroviral compounds that could serve as negative controls and vehicle-only wells that would serve as no-supplement controls
We have demonstrated that the addition of prostaglandin E2 (PGE2) improves lentiviral transduction of human hematopoietic stem and progenitor cells, without evidence of overt integration-site bias in lentivirally transduced cells

Summary

Introduction

Hematopoietic stem cell transplantation is a potentially curative therapy for multiple clinical indications. Promising results with therapeutic applications of lentiviral vector (LVV)-transduced hematopoietic stem cells (HSCs) have been achieved.[1,2,3,4,5] Despite these early successes, it has been challenging to achieve robust and reliable genetic modification of HSCs for all patients and across a variety of therapeutic indications.[6] Overcoming this challenge would expand the therapeutic potential of stem cell-based gene therapy, in disorders where a high level of transgenic expression is required. Approaches to improve lentiviral transduction of HSCs (CD34+ cells) have included soluble factors or gene modulation strategies intended to overcome transduction resistance, including modulation of p21 expression, modulation of mTOR activity, and relief of early capsid-dependent barriers to transduction.[10,11,12] to date, no strategies for increasing LVV transduction efficiency had proven to be sufficiently robust to be brought into the clinic for gene therapy of hematopoietic disorders

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