title: Long-Term Prevention of Congenital Thrombotic Thrombocytopenic Purpura in ADAMTS13 Knockout Mice by Sleeping Beauty Transposon-Mediated Gene Therapy.
abstract: Severe deficiency in the von Willebrand factor-cleaving protease ADAMTS13 (a disintegrin and metalloproteinase with thrombospondin type 1 motif, member 13) because of mutations in theADAMTS13gene can lead to acute episodes of congenital thrombotic thrombocytopenic purpura (TTP), requiring prompt treatment. Current treatment consists of therapeutic or prophylactic infusions of fresh frozen plasma. However, lifelong treatment with plasma products is a stressful therapy for TTP patients. Here, we describe the use of the nonviral sleeping beauty (SB) transposon system as a gene therapeutic approach to realize lifelong expression of ADAMTS13 and subsequent protection against congenital TTP.
  
  
  
  We demonstrated that hydrodynamic tail vein injection of the SB100X system expressing murine ADAMTS13 inAdamts13-/-mice resulted in long-term expression of supraphysiological levels of transgene ADAMTS13 over a period of 25 weeks. Stably expressed ADAMTS13 efficiently removed the prothrombotic ultralarge von Willebrand factor multimers present in the circulation ofAdamts13-/-mice. Moreover, mice stably expressing ADAMTS13 were protected against TTP. The treated mice did not develop severe thrombocytopenia or did organ damage occur when triggered with recombinant von Willebrand factor, and this up to 20 weeks after gene transfer.
  
  
  
  These data demonstrate the feasibility of using SB100X-mediated gene therapy to achieve sustained expression of transgene ADAMTS13 and long-term prophylaxis against TTP inAdamts13-/-mice.
authors.affiliation: From the Laboratory for Thrombosis Research, KU Leuven Campus Kulak Kortrijk, Belgium (S.V., N.V., I.P., H.D., S.F.D.M., K.V.); Mobile DNA, Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany (Z.I.); and Shire, Gene Therapy, Vienna, Austria (H.R.).
authors.forename: Sebastien
authors.initials: S
authors.lastname: Verhenne
authors.affiliation: From the Laboratory for Thrombosis Research, KU Leuven Campus Kulak Kortrijk, Belgium (S.V., N.V., I.P., H.D., S.F.D.M., K.V.); Mobile DNA, Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany (Z.I.); and Shire, Gene Therapy, Vienna, Austria (H.R.).
authors.forename: Nele
authors.initials: N
authors.lastname: Vandeputte
authors.affiliation: From the Laboratory for Thrombosis Research, KU Leuven Campus Kulak Kortrijk, Belgium (S.V., N.V., I.P., H.D., S.F.D.M., K.V.); Mobile DNA, Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany (Z.I.); and Shire, Gene Therapy, Vienna, Austria (H.R.).
authors.forename: Inge
authors.initials: I
authors.lastname: Pareyn
authors.affiliation: From the Laboratory for Thrombosis Research, KU Leuven Campus Kulak Kortrijk, Belgium (S.V., N.V., I.P., H.D., S.F.D.M., K.V.); Mobile DNA, Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany (Z.I.); and Shire, Gene Therapy, Vienna, Austria (H.R.).
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Izsvák
authors.affiliation: From the Laboratory for Thrombosis Research, KU Leuven Campus Kulak Kortrijk, Belgium (S.V., N.V., I.P., H.D., S.F.D.M., K.V.); Mobile DNA, Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany (Z.I.); and Shire, Gene Therapy, Vienna, Austria (H.R.).
authors.forename: Hanspeter
authors.initials: H
authors.lastname: Rottensteiner
authors.affiliation: From the Laboratory for Thrombosis Research, KU Leuven Campus Kulak Kortrijk, Belgium (S.V., N.V., I.P., H.D., S.F.D.M., K.V.); Mobile DNA, Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany (Z.I.); and Shire, Gene Therapy, Vienna, Austria (H.R.).
authors.forename: Hans
authors.initials: H
authors.lastname: Deckmyn
authors.affiliation: From the Laboratory for Thrombosis Research, KU Leuven Campus Kulak Kortrijk, Belgium (S.V., N.V., I.P., H.D., S.F.D.M., K.V.); Mobile DNA, Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany (Z.I.); and Shire, Gene Therapy, Vienna, Austria (H.R.).
authors.forename: Simon F
authors.initials: SF
authors.lastname: De Meyer
authors.affiliation: From the Laboratory for Thrombosis Research, KU Leuven Campus Kulak Kortrijk, Belgium (S.V., N.V., I.P., H.D., S.F.D.M., K.V.); Mobile DNA, Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany (Z.I.); and Shire, Gene Therapy, Vienna, Austria (H.R.). Karen.Vanhoorelbeke@kuleuven.be.
authors.forename: Karen
authors.initials: K
authors.lastname: Vanhoorelbeke
mesh_terms: D000071120:ADAMTS13 Protein / Q000172:deficiency* / Q000235:genetics; D000818:Animals; D004251:DNA Transposable Elements; D004195:Disease Models, Animal; D005240:Feasibility Studies; D020022:Genetic Predisposition to Disease; D015316:Genetic Therapy / Q000379:methods*; D057507:Mice, 129 Strain; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010641:Phenotype; D011697:Purpura, Thrombotic Thrombocytopenic / Q000201:enzymology / Q000235:genetics / Q000517:prevention & control*; D013997:Time Factors; D019895:Transposases / Q000235:genetics*; D014841:von Willebrand Factor
chemical_list: D004251:DNA Transposable Elements; D014841:von Willebrand Factor; D019895:Transposases; C494471:sleeping beauty transposase, human; C502939:ADAMTS13 protein, mouse; D000071120:ADAMTS13 Protein
keywords: ADAMTS13; congenital TTP; gene therapy; sleeping beauty transposon system; von Willebrand factor