title: Long-term reversal of chronic anemia using a hypoxia-regulated erythropoietin gene therapy.
abstract: Anemia is a common clinical problem, and there is much interest in its role in promoting left ventricular hypertrophy through increasing cardiac workload. Normally, red blood cell production is adjusted through the regulation of erythropoietin (Epo) production by the kidney. One important cause of anemia is relative deficiency of Epo, which occurs in most types of renal disease. Clinically, this can be corrected by supplementation with recombinant Epo. Here we describe an oxygen-regulated gene therapy approach to treating homozygous erythropoietin-SV40 T antigen (Epo-TAg(h)) mice with relative erythropoietin deficiency. We used vectors in which murine Epo expression was directed by an Oxford Biomedica hypoxia response element (OBHRE) or a constitutive cytomegalovirus (CMV) promoter. Both corrected anemia, but CMV-Epo-treated mice acquired fatal polycythemia. In contrast, OBHRE-Epo corrected the hematocrit level in anemic mice to a normal physiologic level that stabilized without resulting in polycythemia. Importantly, the OBHRE-Epo vector had no significant effect on the hematocrit of control mice. Homozygous Epo-TAg(h) mice display cardiac hypertrophy, a common adaptive response in patients with chronic anemia. In the OBHRE-Epo-treated Epo-TAg(h) mice, we observed a significant reversal of cardiac hypertrophy. We conclude that the OBHRE promoter gives rise to physiologically regulated Epo secretion such that the hematocrit level is corrected to healthy in anemic Epo-TAg(h) mice. This establishes that a hypoxia regulatory mechanism similar to the natural mechanism can be achieved, and it makes EPO gene therapy more attractive and safer in clinical settings. We envisage that this control system will allow regulated delivery of therapeutic gene products in other ischemic settings.
authors.affiliation: Oxford BioMedica (UK) Ltd; Molecular Immunology Unit, Institute of Child Health, London, United Kingdom. k.binley@oxfordbiomedica.co.uk
authors.forename: Katie
authors.initials: K
authors.lastname: Binley
authors.forename: Zoe
authors.initials: Z
authors.lastname: Askham
authors.forename: Sharifah
authors.initials: S
authors.lastname: Iqball
authors.forename: Hayley
authors.initials: H
authors.lastname: Spearman
authors.forename: Leigh
authors.initials: L
authors.lastname: Martin
authors.forename: Mahesh
authors.initials: M
authors.lastname: de Alwis
authors.forename: Adrian J
authors.initials: AJ
authors.lastname: Thrasher
authors.forename: Robin R
authors.initials: RR
authors.lastname: Ali
authors.forename: Patrick H
authors.initials: PH
authors.lastname: Maxwell
authors.forename: Susan
authors.initials: S
authors.lastname: Kingsman
authors.forename: Stuart
authors.initials: S
authors.lastname: Naylor
mesh_terms: D000740:Anemia / Q000628:therapy*; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D002460:Cell Line; D004921:Erythropoietin / Q000032:analysis / Q000235:genetics* / Q000627:therapeutic use; D005786:Gene Expression Regulation; D015316:Genetic Therapy; D005822:Genetic Vectors; D000860:Hypoxia; D007668:Kidney / Q000502:physiology; D008213:Lymphocyte Activation; D051379:Mice; D008822:Mice, Transgenic; D016133:Polymerase Chain Reaction; D013539:Simian virus 40 / Q000235:genetics; D013154:Spleen / Q000276:immunology; D014162:Transfection
chemical_list: D000952:Antigens, Polyomavirus Transforming; D004921:Erythropoietin