title: Gene transfer of arginine kinase to skeletal muscle using adeno-associated virus.
abstract: In this study, we tested the feasibility of non-invasively measuring phosphoarginine (PArg) after gene delivery of arginine kinase (AK) using an adeno-associated virus (AAV) to murine hindlimbs. This was achieved by evaluating the time course, regional distribution and metabolic flux of PArg using (31)phosphorus magnetic resonance spectroscopy ((31)P-MRS). AK gene was injected into the gastrocnemius of the left hindlimb of C57Bl10 mice (age 5 weeks, male) using self-complementary AAV, type 2/8 with desmin promoter. Non-localized (31)P-MRS data were acquired over 9 months after injection using 11.1-T and 17.6-T Bruker Avance spectrometers. In addition, (31)P two-dimensional chemical shift imaging and saturation transfer experiments were performed to examine the spatial distribution and metabolic flux of PArg, respectively. PArg was evident in each injected mouse hindlimb after gene delivery, increased until 28 weeks, and remained elevated for at least 9 months (P<0.05). Furthermore, PArg was primarily localized to the injected posterior hindimb region and the metabolite was in exchange with ATP. Overall, the results show the viability of AAV gene transfer of AK gene to skeletal muscle, and provide support of PArg as a reporter that can be used to non-invasively monitor the transduction of genes for therapeutic interventions.
authors.affiliation: Department of Physical Therapy, University of Florida, Gainesville, FL, USA.
authors.forename: S C
authors.initials: SC
authors.lastname: Forbes
authors.affiliation: Department of Physiology, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: L T
authors.initials: LT
authors.lastname: Bish
authors.affiliation: Department of Physical Therapy, University of Florida, Gainesville, FL, USA.
authors.forename: F
authors.initials: F
authors.lastname: Ye
authors.affiliation: Department of Anatomy and Cell Biology, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: J
authors.initials: J
authors.lastname: Spinazzola
authors.affiliation: Department of Physiology and Functional Genomics, University of Florida, Gainesville, FL, USA.
authors.forename: C
authors.initials: C
authors.lastname: Baligand
authors.affiliation: Advanced Magnetic Resonance Imaging and Spectroscopy Facility, University of Florida, Gainesville, FL, USA.
authors.forename: D
authors.initials: D
authors.lastname: Plant
authors.affiliation: Department of Physical Therapy, University of Florida, Gainesville, FL, USA.
authors.forename: K
authors.initials: K
authors.lastname: Vandenborne
authors.affiliation: Department of Anatomy and Cell Biology, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: E R
authors.initials: ER
authors.lastname: Barton
authors.affiliation: Department of Physiology, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: H L
authors.initials: HL
authors.lastname: Sweeney
authors.affiliation: Department of Physiology and Functional Genomics, University of Florida, Gainesville, FL, USA.
authors.forename: G A
authors.initials: GA
authors.lastname: Walter
mesh_terms: D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives* / Q000235:genetics / Q000378:metabolism; D001122:Arginine Kinase / Q000235:genetics* / Q000627:therapeutic use; D000229:Dependovirus / Q000235:genetics*; D015316:Genetic Therapy; D005822:Genetic Vectors; D006614:Hindlimb / Q000378:metabolism; D051379:Mice; D018482:Muscle, Skeletal / Q000378:metabolism; D009943:Organophosphorus Compounds / Q000378:metabolism; D011401:Promoter Regions, Genetic; D014161:Transduction, Genetic
chemical_list: D009943:Organophosphorus Compounds; C015441:phospho-L-arginine; D001120:Arginine; D001122:Arginine Kinase