PMID- 31660421 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20240214 IS - 2329-0501 (Print) IS - 2329-0501 (Electronic) IS - 2329-0501 (Linking) VI - 15 DP - 2019 Dec 13 TI - AAV Gene Therapy Utilizing Glycosylation-Independent Lysosomal Targeting Tagged GAA in the Hypoglossal Motor System of Pompe Mice. PG - 194-203 LID - 10.1016/j.omtm.2019.08.009 [doi] AB - Pompe disease is caused by mutations in the gene encoding the lysosomal glycogen-metabolizing enzyme, acid-alpha glucosidase (GAA). Tongue myofibers and hypoglossal motoneurons appear to be particularly susceptible in Pompe disease. Here we used intramuscular delivery of adeno-associated virus serotype 9 (AAV9) for targeted delivery of an enhanced form of GAA to tongue myofibers and motoneurons in 6-month-old Pompe (Gaa (-/-) ) mice. We hypothesized that addition of a glycosylation-independent lysosomal targeting tag to the protein would result in enhanced expression in tongue (hypoglossal) motoneurons when compared to the untagged GAA. Mice received an injection into the base of the tongue with AAV9 encoding either the tagged or untagged enzyme; tissues were harvested 4 months later. Both AAV9 constructs effectively drove GAA expression in lingual myofibers and hypoglossal motoneurons. However, mice treated with the AAV9 construct encoding the modified GAA enzyme had a >200% increase in the number of GAA-positive motoneurons as compared to the untagged GAA (p < 0.008). Our results confirm that tongue delivery of AAV9-encoding GAA can effectively target tongue myofibers and associated motoneurons in Pompe mice and indicate that the effectiveness of this approach can be improved by addition of the glycosylation-independent lysosomal targeting tag. CI - (c) 2019 The Authors. FAU - Doyle, Brendan M AU - Doyle BM AD - Department of Physical Therapy, University of Florida, Gainesville, FL 32610, USA. AD - Center for Respiratory Research and Rehabilitation, University of Florida, Gainesville, FL 32610, USA. FAU - Turner, Sara M F AU - Turner SMF AD - Department of Physical Therapy, University of Florida, Gainesville, FL 32610, USA. AD - Center for Respiratory Research and Rehabilitation, University of Florida, Gainesville, FL 32610, USA. FAU - Sunshine, Michael D AU - Sunshine MD AD - Department of Physical Therapy, University of Florida, Gainesville, FL 32610, USA. AD - Center for Respiratory Research and Rehabilitation, University of Florida, Gainesville, FL 32610, USA. FAU - Doerfler, Phillip A AU - Doerfler PA AD - Department of Pediatrics, University of Florida, Gainesville, FL 32610, USA. AD - Powell Gene Therapy Center, University of Florida, Gainesville, FL 32610, USA. FAU - Poirier, Amy E AU - Poirier AE AD - Department of Neuroscience, University of Florida, Gainesville, FL 32610, USA. AD - Center for Respiratory Research and Rehabilitation, University of Florida, Gainesville, FL 32610, USA. FAU - Vaught, Lauren A AU - Vaught LA AD - Department of Pediatrics, University of Florida, Gainesville, FL 32610, USA. AD - Powell Gene Therapy Center, University of Florida, Gainesville, FL 32610, USA. AD - Center for Respiratory Research and Rehabilitation, University of Florida, Gainesville, FL 32610, USA. FAU - Jorgensen, Marda L AU - Jorgensen ML AD - Department of Pediatrics, University of Florida, Gainesville, FL 32610, USA. FAU - Falk, Darin J AU - Falk DJ AD - Department of Pediatrics, University of Florida, Gainesville, FL 32610, USA. AD - Powell Gene Therapy Center, University of Florida, Gainesville, FL 32610, USA. AD - Center for Respiratory Research and Rehabilitation, University of Florida, Gainesville, FL 32610, USA. FAU - Byrne, Barry J AU - Byrne BJ AD - Department of Pediatrics, University of Florida, Gainesville, FL 32610, USA. AD - Powell Gene Therapy Center, University of Florida, Gainesville, FL 32610, USA. AD - Center for Respiratory Research and Rehabilitation, University of Florida, Gainesville, FL 32610, USA. FAU - Fuller, David D AU - Fuller DD AD - Department of Physical Therapy, University of Florida, Gainesville, FL 32610, USA. AD - Mcknight Brain Institute, University of Florida, Gainesville, FL 32610, USA. AD - Center for Respiratory Research and Rehabilitation, University of Florida, Gainesville, FL 32610, USA. LA - eng GR - F31 HL145831/HL/NHLBI NIH HHS/United States GR - K01 AR066077/AR/NIAMS NIH HHS/United States GR - R01 HD052682/HD/NICHD NIH HHS/United States GR - T32 HD043730/HD/NICHD NIH HHS/United States PT - Journal Article DEP - 20190831 PL - United States TA - Mol Ther Methods Clin Dev JT - Molecular therapy. Methods & clinical development JID - 101624857 PMC - PMC6807287 OTO - NOTNLM OT - AAV OT - GAA OT - Pompe OT - gene therapy OT - genioglossus OT - hypoglossal OT - lysosomal storage disease OT - motoneuron OT - respiratory OT - tongue EDAT- 2019/10/30 06:00 MHDA- 2019/10/30 06:01 PMCR- 2019/08/31 CRDT- 2019/10/30 06:00 PHST- 2019/06/06 00:00 [received] PHST- 2019/08/23 00:00 [accepted] PHST- 2019/10/30 06:00 [entrez] PHST- 2019/10/30 06:00 [pubmed] PHST- 2019/10/30 06:01 [medline] PHST- 2019/08/31 00:00 [pmc-release] AID - S2329-0501(19)30089-0 [pii] AID - 10.1016/j.omtm.2019.08.009 [doi] PST - epublish SO - Mol Ther Methods Clin Dev. 2019 Aug 31;15:194-203. doi: 10.1016/j.omtm.2019.08.009. eCollection 2019 Dec 13.