Open AccessEnhanced Genetic Modification of Adult Growth Factor Mobilized Peripheral Blood Hematopoietic Stem and Progenitor Cells With Rapamycin
Author(s) -
Li Lijing,
Torres-Coronado Mónica,
Gu Angel,
Rao Anitha,
Gardner Agnes M.,
Epps Elizabeth W.,
Gonzalez Nancy,
Tran Chy-Anh,
Wu Xiwei,
Wang Jin-Hui,
DiGiusto David L.
Publication year - 2014
Publication title -
stem cells translational medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.781
H-Index - 71
eISSN - 2157-6580
pISSN - 2157-6564
DOI - 10.5966/sctm.2014-0010
Subject(s) - progenitor cell , haematopoiesis , biology , genetic enhancement , clonogenic assay , stem cell , transduction (biophysics) , cd34 , viral vector , myeloid , microbiology and biotechnology , transgene , population , cancer research , immunology , gene , cell , genetics , medicine , recombinant dna , biochemistry , environmental health
Genetic modification of adult human hematopoietic stem and progenitor cells (HSPCs) with lentiviral vectors leads to long‐term gene expression in the progeny of the HSPCs and has been used to successfully treat several monogenic diseases. In some cases, the gene‐modified cells have a selective growth advantage over nonmodified cells and eventually are the dominant engrafted population. However, in disease indications for which the gene‐modified cells do not have a selective advantage, optimizing transduction of HSPC is paramount to successful stem cell‐based gene therapy. We demonstrate here that transduction of adult CD34+ HSPCs with lentiviral vectors in the presence of rapamycin, a widely used mTORC1 inhibitor, results in an approximately threefold increase in stable gene marking with minimal effects on HSPC growth and differentiation. Using this approach, we have demonstrated that we can enhance the frequency of gene‐modified HSPCs that give rise to clonogenic progeny in vitro without excessive increases in the number of vector copies per cell or changes in integration pattern. The genetic marking of HSPCs and expression of transgenes is durable, and transplantation of gene‐modified HSPCs into immunodeficient mice results in high levels of gene marking of the lymphoid and myeloid progeny in vivo. The prior safe clinical history of rapamycin in other applications supports the use of this compound to generate gene‐modified autologous HSPCs for our HIV gene therapy clinical trials.