Integrating HDAd5/35++ Vectors as a New Platform for HSC Gene Therapy of Hemoglobinopathies
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)-mg...
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doaj-3f0562ba57da41779a3d7d4fd142f2812020-11-25T01:01:15ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012018-06-019142152Integrating HDAd5/35++ Vectors as a New Platform for HSC Gene Therapy of HemoglobinopathiesChang Li0Nikoletta Psatha1Hongjie Wang2Manvendra Singh3Himanshu Bhusan Samal4Wenli Zhang5Anja Ehrhardt6Zsuzsanna Izsvák7Thalia Papayannopoulou8André Lieber9Division of Medical Genetics, Department of Medicine, University of Washington, Box 357720, Seattle, WA 98195, USADivision of Hematology Department of Medicine, University of Washington, Seattle, WA 98195, USADivision of Medical Genetics, Department of Medicine, University of Washington, Box 357720, Seattle, WA 98195, USAMax-Delbrück-Center for Molecular Medicine, Berlin, 13092 GermanyMax-Delbrück-Center for Molecular Medicine, Berlin, 13092 GermanyWitten/Herdecke University, Witten, 58448, GermanyWitten/Herdecke University, Witten, 58448, GermanyMax-Delbrück-Center for Molecular Medicine, Berlin, 13092 GermanyDivision of Hematology Department of Medicine, University of Washington, Seattle, WA 98195, USADivision of Medical Genetics, Department of Medicine, University of Washington, Box 357720, Seattle, WA 98195, USA; Department of Pathology, University of Washington, Seattle, WA 98195, USA; Corresponding author: André Lieber, University of Washington, Box 357720, Seattle, WA 98195, USA.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 allow for efficient transduction of long-term repopulating HSCs and high-level, almost pancellular γ-globin expression in erythrocytes. Furthermore, our HDAd5/35++ vectors have a larger insert capacity and a safer integration pattern than currently used lentivirus vectors. Keywords: hematopoietic stem cells, Sleeping Beauty transposase, beta-thalassemia, sickle cell disease, gamma globin, integration, gene additionhttp://www.sciencedirect.com/science/article/pii/S2329050118300160 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Chang Li Nikoletta Psatha Hongjie Wang Manvendra Singh Himanshu Bhusan Samal Wenli Zhang Anja Ehrhardt Zsuzsanna Izsvák Thalia Papayannopoulou André Lieber |
spellingShingle |
Chang Li Nikoletta Psatha Hongjie Wang Manvendra Singh Himanshu Bhusan Samal Wenli Zhang Anja Ehrhardt Zsuzsanna Izsvák Thalia Papayannopoulou André Lieber Integrating HDAd5/35++ Vectors as a New Platform for HSC Gene Therapy of Hemoglobinopathies Molecular Therapy: Methods & Clinical Development |
author_facet |
Chang Li Nikoletta Psatha Hongjie Wang Manvendra Singh Himanshu Bhusan Samal Wenli Zhang Anja Ehrhardt Zsuzsanna Izsvák Thalia Papayannopoulou André Lieber |
author_sort |
Chang Li |
title |
Integrating HDAd5/35++ Vectors as a New Platform for HSC Gene Therapy of Hemoglobinopathies |
title_short |
Integrating HDAd5/35++ Vectors as a New Platform for HSC Gene Therapy of Hemoglobinopathies |
title_full |
Integrating HDAd5/35++ Vectors as a New Platform for HSC Gene Therapy of Hemoglobinopathies |
title_fullStr |
Integrating HDAd5/35++ Vectors as a New Platform for HSC Gene Therapy of Hemoglobinopathies |
title_full_unstemmed |
Integrating HDAd5/35++ Vectors as a New Platform for HSC Gene Therapy of Hemoglobinopathies |
title_sort |
integrating hdad5/35++ vectors as a new platform for hsc gene therapy of hemoglobinopathies |
publisher |
Elsevier |
series |
Molecular Therapy: Methods & Clinical Development |
issn |
2329-0501 |
publishDate |
2018-06-01 |
description |
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 allow for efficient transduction of long-term repopulating HSCs and high-level, almost pancellular γ-globin expression in erythrocytes. Furthermore, our HDAd5/35++ vectors have a larger insert capacity and a safer integration pattern than currently used lentivirus vectors. Keywords: hematopoietic stem cells, Sleeping Beauty transposase, beta-thalassemia, sickle cell disease, gamma globin, integration, gene addition |
url |
http://www.sciencedirect.com/science/article/pii/S2329050118300160 |
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