title: Stable human FIX expression after 0.9G intrauterine gene transfer of self-complementary adeno-associated viral vector 5 and 8 in macaques.
abstract: Intrauterine gene transfer (IUGT) offers ontological advantages including immune naiveté mediating tolerance to the vector and transgenic products, and effecting a cure before development of irreversible pathology. Despite proof-of-principle in rodent models, expression efficacy with a therapeutic transgene has yet to be demonstrated in a preclinical nonhuman primate (NHP) model. We aimed to determine the efficacy of human Factor IX (hFIX) expression after adeno-associated-viral (AAV)-mediated IUGT in NHP. We injected 1.0-1.95 × 10(13) vector genomes (vg)/kg of self-complementary (sc) AAV5 and 8 with a LP1-driven hFIX transgene intravenously in 0.9G late gestation NHP fetuses, leading to widespread transduction with liver tropism. Liver-specific hFIX expression was stably maintained between 8 and 112% of normal activity in injected offspring followed up for 2-22 months. AAV8 induced higher hFIX expression (P = 0.005) and milder immune response than AAV5. Random hepatocellular integration was found with no hotspots. Transplacental spread led to low-level maternal tissue transduction, without evidence of immunotoxicity or germline transduction in maternal oocytes. A single intravenous injection of scAAV-LP1-hFIXco to NHP fetuses in late-gestation produced sustained clinically-relevant levels of hFIX with liver-specific expression and a non-neutralizing immune response. These data are encouraging for conditions where gene transfer has the potential to avert perinatal death and long-term irreversible sequelae.
authors.affiliation: Department of Obstetrics and Gynaecology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Citra N Z
authors.initials: CN
authors.lastname: Mattar
authors.forename: Amit C
authors.initials: AC
authors.lastname: Nathwani
authors.forename: Simon N
authors.initials: SN
authors.lastname: Waddington
authors.forename: Niraja
authors.initials: N
authors.lastname: Dighe
authors.forename: Christine
authors.initials: C
authors.lastname: Kaeppel
authors.forename: Ali
authors.initials: A
authors.lastname: Nowrouzi
authors.forename: Jenny
authors.initials: J
authors.lastname: Mcintosh
authors.forename: Nuryanti B
authors.initials: NB
authors.lastname: Johana
authors.forename: Bryan
authors.initials: B
authors.lastname: Ogden
authors.forename: Nicholas M
authors.initials: NM
authors.lastname: Fisk
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Davidoff
authors.forename: Anna
authors.initials: A
authors.lastname: David
authors.forename: Donald
authors.initials: D
authors.lastname: Peebles
authors.forename: Marcus B
authors.initials: MB
authors.lastname: Valentine
authors.forename: Jens-Uwe
authors.initials: JU
authors.lastname: Appelt
authors.forename: Christof
authors.initials: C
authors.lastname: von Kalle
authors.forename: Manfred
authors.initials: M
authors.lastname: Schmidt
authors.forename: Arijit
authors.initials: A
authors.lastname: Biswas
authors.forename: Mahesh
authors.initials: M
authors.lastname: Choolani
authors.forename: Jerry K Y
authors.initials: JK
authors.lastname: Chan
mesh_terms: D000818:Animals; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics* / Q000276:immunology; D005164:Factor IX / Q000235:genetics*; D005260:Female; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics* / Q000493:pharmacokinetics; D057809:HEK293 Cells; D002836:Hemophilia B / Q000235:genetics / Q000628:therapy*; D006801:Humans; D007267:Injections; D008252:Macaca fascicularis; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy; D014161:Transduction, Genetic; D016662:Virus Integration
chemical_list: D005164:Factor IX