title: In vivo genome editing of the albumin locus as a platform for protein replacement therapy.
abstract: Site-specific genome editing provides a promising approach for achieving long-term, stable therapeutic gene expression. Genome editing has been successfully applied in a variety of preclinical models, generally focused on targeting the diseased locus itself; however, limited targeting efficiency or insufficient expression from the endogenous promoter may impede the translation of these approaches, particularly if the desired editing event does not confer a selective growth advantage. Here we report a general strategy for liver-directed protein replacement therapies that addresses these issues: zinc finger nuclease (ZFN) -mediated site-specific integration of therapeutic transgenes within the albumin gene. By using adeno-associated viral (AAV) vector delivery in vivo, we achieved long-term expression of human factors VIII and IX (hFVIII and hFIX) in mouse models of hemophilia A and B at therapeutic levels. By using the same targeting reagents in wild-type mice, lysosomal enzymes were expressed that are deficient in Fabry and Gaucher diseases and in Hurler and Hunter syndromes. The establishment of a universal nuclease-based platform for secreted protein production would represent a critical advance in the development of safe, permanent, and functional cures for diverse genetic and nongenetic diseases.
authors: Division of Hematology, Children's Hospital of Philadelphia, Philadelphia, PA;
authors: Rajiv
authors: http://orcid.org/0000-0002-8147-5723
authors: R
authors: Sharma
authors: Division of Hematology, Children's Hospital of Philadelphia, Philadelphia, PA; Howard Hughes Medical Institute, Philadelphia, PA; and.
authors: Xavier M
authors: http://orcid.org/0000-0003-2430-8598
authors: XM
authors: Anguela
authors: Sangamo BioSciences, Richmond, CA.
authors: Yannick
authors: Y
authors: Doyon
authors: Sangamo BioSciences, Richmond, CA.
authors: Thomas
authors: T
authors: Wechsler
authors: Sangamo BioSciences, Richmond, CA.
authors: Russell C
authors: RC
authors: DeKelver
authors: Sangamo BioSciences, Richmond, CA.
authors: Scott
authors: S
authors: Sproul
authors: Sangamo BioSciences, Richmond, CA.
authors: David E
authors: DE
authors: Paschon
authors: Sangamo BioSciences, Richmond, CA.
authors: Jeffrey C
authors: JC
authors: Miller
authors: Division of Hematology, Children's Hospital of Philadelphia, Philadelphia, PA;
authors: Robert J
authors: RJ
authors: Davidson
authors: Sangamo BioSciences, Richmond, CA.
authors: David
authors: D
authors: Shivak
authors: Division of Hematology, Children's Hospital of Philadelphia, Philadelphia, PA;
authors: Shangzhen
authors: S
authors: Zhou
authors: Division of Hematology, Children's Hospital of Philadelphia, Philadelphia, PA;
authors: Julianne
authors: J
authors: Rieders
authors: Sangamo BioSciences, Richmond, CA.
authors: Philip D
authors: PD
authors: Gregory
authors: Sangamo BioSciences, Richmond, CA.
authors: Michael C
authors: MC
authors: Holmes
authors: Sangamo BioSciences, Richmond, CA.
authors: Edward J
authors: EJ
authors: Rebar
authors: Division of Hematology, Children's Hospital of Philadelphia, Philadelphia, PA; Howard Hughes Medical Institute, Philadelphia, PA; and.
authors: Katherine A
authors: KA
authors: High
mesh_terms: D000418:Albumins / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D000229:Dependovirus / Q000235:genetics; D004720:Endonucleases; D056947:Enzyme Replacement Therapy; D000795:Fabry Disease / Q000235:genetics / Q000628:therapy; D005164:Factor IX / Q000235:genetics; D005169:Factor VIII / Q000235:genetics; D005776:Gaucher Disease / Q000235:genetics / Q000628:therapy; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage; D016678:Genome; D006467:Hemophilia A / Q000235:genetics / Q000628:therapy; D002836:Hemophilia B / Q000235:genetics / Q000628:therapy; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008099:Liver / Q000378:metabolism*; D008247:Lysosomes / Q000201:enzymology; D051379:Mice; D008810:Mice, Inbred C57BL; D008059:Mucopolysaccharidosis I / Q000235:genetics / Q000628:therapy; D016532:Mucopolysaccharidosis II / Q000235:genetics / Q000628:therapy; D011401:Promoter Regions, Genetic / Q000235:genetics; D017393:RNA Editing; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019076:Transgenes / Q000502:physiology*; D016335:Zinc Fingers
chemical_list: D000418:Albumins; D012333:RNA, Messenger; C078147:F8 protein, human; D005169:Factor VIII; D005164:Factor IX; D004720:Endonucleases