Sleeping Beauty-Mediated Drug Resistance Gene Transfer in Human Hematopoietic Progenitor Cells

• Published in: Human gene therapy Vol. 26; no. 10; p. 657
• Main Authors: Hyland, Kendra A, Olson, Erik R, McIvor, R Scott
• Format: Journal Article
• Language: English
• Published: United States 01.10.2015
• Subjects: Antigens, CD34 - genetics; Dipyridamole - administration & dosage; DNA Transposable Elements - genetics; Drug Resistance - drug effects; Drug Resistance - genetics; Gene Transfer Techniques; Genetic Therapy; Green Fluorescent Proteins - genetics; Hematopoietic Stem Cells; Humans; Methotrexate - administration & dosage; Tetrahydrofolate Dehydrogenase - genetics
• ISSN: 1557-7422
• DOI: 10.1089/hum.2015.058

The Sleeping Beauty (SB) transposon system can insert sequences into mammalian chromosomes, supporting long-term expression of both reporter and therapeutic genes. Hematopoietic progenitor cells (HPCs) are an ideal therapeutic gene transfer target as they are used in therapy for a variety of hematologic and metabolic conditions. As successful SB-mediated gene transfer into human CD34(+) HPCs has been reported by several laboratories, we sought to extend these studies to the introduction of a therapeutic gene conferring resistance to methotrexate (MTX), potentially providing a chemoprotective effect after engraftment. SB-mediated transposition of hematopoietic progenitors, using a transposon encoding an L22Y variant dihydrofolate reductase fused to green fluorescent protein, conferred resistance to methotrexate and dipyridamole, a nucleoside transport inhibitor that tightens MTX selection conditions, as assessed by in vitro hematopoietic colony formation. Transposition of individual transgenes was confirmed by sequence analysis of transposon-chromosome junctions recovered by linear amplification-mediated PCR. These studies demonstrate the potential of SB-mediated transposition of HPCs for expression of drug resistance genes for selective and chemoprotective applications.