Integrating HDAd5/35++ Vectors as a New Platform for HSC Gene Therapy of Hemoglobinopathies

Molecular Therapy. Methods & Clinical Development
Chang LiAndré Lieber

Abstract

We generated an integrating, CD46-targeted, helper-dependent adenovirus HDAd5/35++ vector system for hematopoietic stem cell (HSC) gene therapy. The ∼12-kb transgene cassette included a β-globin locus control region (LCR)/promoter driven human γ-globin gene and an elongation factor alpha-1 (EF1α)-mgmtP140K expression cassette, which allows for drug-controlled increase of γ-globin-expressing erythrocytes. We transduced bone marrow lineage-depleted cells from human CD46-transgenic mice and transplanted them into lethally irradiated recipients. The percentage of γ-globin-positive cells in peripheral blood erythrocytes in primary and secondary transplant recipients was stable and greater than 90%. The γ-globin level was 10%-20% of adult mouse globin. Transgene integration, mediated by a hyperactive Sleeping Beauty SB100x transposase, was random, without a preference for genes. A second set of studies was performed with peripheral blood CD34+ cells from mobilized donors. 10 weeks after transplantation of transduced cells, human cells were harvested from the bone marrow and differentiated ex vivo into erythroid cells. Erythroid cells expressed γ-globin at a level of 20% of adult α-globin. Our studies suggest that HDAd35++ vectors all...Continue Reading

Citations

Sep 12, 2018·Human Gene Therapy·Marina Cavazzana, Fulvio Mavilio
Nov 14, 2018·The Journal of Clinical Investigation·Hongjie WangAndré Lieber
Nov 11, 2019·FEBS Letters·Chang Li, André Lieber
Nov 28, 2019·Viruses·Yogendra Singh RajawatHans-Peter Kiem
Sep 20, 2020·Molecular Therapy : the Journal of the American Society of Gene Therapy·Chang LiAndré Lieber
Sep 13, 2021·Molecular Therapy : the Journal of the American Society of Gene Therapy·Byung-Chul LeeCynthia E Dunbar

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Methods Mentioned

BETA
Transgenic
flow cytometry
genetic modification
PCR
Illumina sequencing
transfection
FCS
FACS

Software Mentioned

GraphPad
Illumina
GraphPad Prism
FlowJo
Bowtie2
BEDTools
CASAVA
Aevi

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