title: Correction of ADAMTS13 deficiency by in utero gene transfer of lentiviral vector encoding ADAMTS13 genes.
abstract: Deficiency of A Disintegrin And Metalloprotease with ThromboSpondin (ADAMTS13) results in thrombotic thrombocytopenic purpura (TTP). Plasma infusion or exchange is the only effective treatment to date. We show in this study that an administration of a self-inactivating lentiviral vector encoding human full-length ADAMTS13 and a variant truncated after the spacer domain (MDTCS) in mice by in utero injection at embryonic days 8 and 14 resulted in detectable plasma proteolytic activity (approximately 5-70%), which persisted for the length of the study (up to 24 weeks). Intravascular injection via a vitelline vein at E14 was associated with significantly lower rate of fetal loss than intra-amniotic injection, suggesting that the administration of vector at E14 may be a preferred gestational age for vector delivery. The mice expressing ADAMTS13 and MDTCS exhibited reduced sizes of von Willebrand factor (vWF) compared to the Adamts13(-/-) mice expressing enhanced green fluorescent protein (eGFP). Moreover, the mice expressing both ADAMTS13 and MDTCS showed a significant prolongation of ferric chloride-induced carotid arterial occlusion time as compared to the Adamts13(-/-) expressing eGFP. The data demonstrate the successful correction of the prothrombotic phenotypes in Adamts13(-/-) mice by a single in utero injection of lentiviral vectors encoding human ADAMTS13 genes, providing the basis for developing a gene therapy for hereditary TTP in humans.
authors.affiliation: Department of Pathology and Laboratory Medicine, The Children's Hospital of Philadelphia and The University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
authors.forename: Masami
authors.initials: M
authors.lastname: Niiya
authors.forename: Masayuki
authors.initials: M
authors.lastname: Endo
authors.forename: Dezhi
authors.initials: D
authors.lastname: Shang
authors.forename: Philip W
authors.initials: PW
authors.lastname: Zoltick
authors.forename: Nidal E
authors.initials: NE
authors.lastname: Muvarak
authors.forename: Wenjing
authors.initials: W
authors.lastname: Cao
authors.forename: Sheng-Yu
authors.initials: SY
authors.lastname: Jin
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Skipwith
authors.forename: David G
authors.initials: DG
authors.lastname: Motto
authors.forename: Alan W
authors.initials: AW
authors.lastname: Flake
authors.forename: X Long
authors.initials: XL
authors.lastname: Zheng
mesh_terms: D051722:ADAM Proteins / Q000172:deficiency / Q000235:genetics* / Q000502:physiology; D000071120:ADAMTS13 Protein; D000818:Animals; D015153:Blotting, Western; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D047468:Immunoprecipitation; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D008817:Mice, Mutant Strains; D008856:Microscopy, Fluorescence; D011697:Purpura, Thrombotic Thrombocytopenic / Q000235:genetics / Q000628:therapy*; D014599:Uterus
chemical_list: D051722:ADAM Proteins; D000071120:ADAMTS13 Protein; C099604:ADAMTS13 protein, human