title: The β-globin locus control region in combination with the EF1α short promoter allows enhanced lentiviral vector-mediated erythroid gene expression with conserved multilineage activity.
abstract: Some gene therapy strategies are compromised by the levels of gene expression required for therapeutic benefit, and also by the breadth of cell types that require correction. We designed a lentiviral vector system in which a transgene is under the transcriptional control of the short form of constitutively acting elongation factor 1α promoter (EFS) combined with essential elements of the locus control region of the β-globin gene (β-LCR). We show that the β-LCR can upregulate EFS activity specifically in erythroid cells but does not alter EFS activity in myeloid or lymphoid cells. Experiments using the green fluorescent protein (GFP) reporter or the human adenosine deaminase (ADA) gene demonstrate 3-7 times upregulation in vitro but >20 times erythroid-specific upregulation in vivo, the effects of which were sustained for 1 year. The addition of the β-LCR did not alter the mutagenic potential of the vector in in vitro mutagenesis (IM) assays although microarray analysis showed that the β-LCR upregulates ~9% of neighboring genes. This vector design therefore combines the benefits of multilineage gene expression with high-level erythroid expression, and has considerable potential for correction of multisystem diseases including certain lysosomal storage diseases through a hematopoietic stem cell (HSC) gene therapy approach.
authors.affiliation: Centre for Immunodeficiency, Molecular Immunology Unit, Institute of Child Health, University College London, London, UK.
authors.forename: Claudia A
authors.initials: CA
authors.lastname: Montiel-Equihua
authors.forename: Lin
authors.initials: L
authors.lastname: Zhang
authors.forename: Sean
authors.initials: S
authors.lastname: Knight
authors.forename: Heba
authors.initials: H
authors.lastname: Saadeh
authors.forename: Simone
authors.initials: S
authors.lastname: Scholz
authors.forename: Marlene
authors.initials: M
authors.lastname: Carmo
authors.forename: Maria E
authors.initials: ME
authors.lastname: Alonso-Ferrero
authors.forename: Michael P
authors.initials: MP
authors.lastname: Blundell
authors.forename: Aiste
authors.initials: A
authors.lastname: Monkeviciute
authors.forename: Reiner
authors.initials: R
authors.lastname: Schulz
authors.forename: Mary
authors.initials: M
authors.lastname: Collins
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Takeuchi
authors.forename: Manfred
authors.initials: M
authors.lastname: Schmidt
authors.forename: Lynette
authors.initials: L
authors.lastname: Fairbanks
authors.forename: Michael
authors.initials: M
authors.lastname: Antoniou
authors.forename: Adrian J
authors.initials: AJ
authors.lastname: Thrasher
authors.forename: H Bobby
authors.initials: HB
authors.lastname: Gaspar
mesh_terms: D000243:Adenosine Deaminase / Q000235:genetics; D000818:Animals; D002460:Cell Line; D015672:Erythroid Precursor Cells / Q000378:metabolism*; D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D057809:HEK293 Cells; D006412:Hematopoietic Stem Cells; D006801:Humans; D019169:Jurkat Cells; D016086:Lentivirus / Q000235:genetics; D019901:Locus Control Region; D016464:Lysosomal Storage Diseases / Q000235:genetics / Q000628:therapy; D051379:Mice; D008810:Mice, Inbred C57BL; D020648:Peptide Elongation Factor 1 / Q000235:genetics*; D010445:Peptide Elongation Factors / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D020298:U937 Cells; D015854:Up-Regulation; D055544:beta-Globins / Q000235:genetics*
chemical_list: D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D055544:beta-Globins; D049452:Green Fluorescent Proteins; D000243:Adenosine Deaminase