Systemic gene delivery transduces the enteric nervous system of guinea pigs and cynomolgus macaques

Characterization of adeno-associated viral vector (AAV) mediated gene delivery to the enteric nervous system (ENS) was recently described in mice and rats. In these proof-of-concept experiments, we show that intravenous injections of clinically relevant AAVs can transduce the ENS in guinea pigs and...

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Published in:	Gene therapy Vol. 24; no. 10; pp. 640 - 648
Main Authors:	Gombash, S E, Cowley, C J, Fitzgerald, J A, Lepak, C A, Neides, M G, Hook, K, Todd, L J, Wang, G-D, Mueller, C, Kaspar, B K, Bielefeld, E C, Fischer, A J, Wood, J D, Foust, K D
Format:	Journal Article
Language:	English
Published:	London Nature Publishing Group UK 01-10-2017 Nature Publishing Group
Subjects:	14/35 42/34 631/1647/2300 631/378/1959 631/378/1959/1315/1951 631/378/87 692/700/565/201 82/51 82/80 Anatomy Animal tissues Animals Biomedical and Life Sciences Biomedicine Cell Biology Dependovirus - genetics Digestive system Dorsal root ganglia Dosage and administration Enteric nervous system Enteric Nervous System - metabolism Expression vectors Female Ganglia, Spinal - cytology Ganglia, Spinal - metabolism Gastrointestinal tract Gene Expression Gene Therapy Gene transfer Gene Transfer Techniques Genetic modification Genetic Therapy - methods Genetic Vectors - genetics Genetics Genome editing Green fluorescent protein Green fluorescent proteins Green Fluorescent Proteins - genetics Green Fluorescent Proteins - metabolism Guinea Pigs Human Genetics Information processing Injection Injections, Intravenous Intravenous administration Intravenous injections Large intestine Macaca fascicularis Macaques Male Motor nuclei Nanotechnology Neonates Nervous system Neurons Neurons - metabolism Optics original-article Phosphates Primates Rats Rodents Stomach Vagus Nerve - metabolism Guinea
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Abstract	Characterization of adeno-associated viral vector (AAV) mediated gene delivery to the enteric nervous system (ENS) was recently described in mice and rats. In these proof-of-concept experiments, we show that intravenous injections of clinically relevant AAVs can transduce the ENS in guinea pigs and non-human primates. Neonatal guinea pigs were given intravenous injections of either AAV8 or AAV9 vectors that contained a green fluorescent protein (GFP) expression cassette or phosphate-buffered saline. Piglets were euthanized three weeks post injection and tissues were harvested for immunofluorescent analysis. GFP expression was detected in myenteric and submucosal neurons along the length of the gastrointestinal tract in AAV8 injected guinea pigs. GFP-positive neurons were found in dorsal motor nucleus of the vagus and dorsal root ganglia. Less transduction occurred in AAV9-treated tissues. Gastrointestinal tissues were analyzed from young cynomolgus macaques that received systemic injection of AAV9 GFP. GFP expression was detected in myenteric neurons of the stomach, small and large intestine. These data demonstrate that ENS gene delivery translates to larger species. This work develops tools for the field of neurogastroenterology to explore gut physiology and anatomy using emerging technologies such as optogenetics and gene editing. It also provides a basis to develop novel therapies for chronic gut disorders.
AbstractList	Characterization of adeno-associated viral vector (AAV) mediated gene delivery to the enteric nervous system (ENS) was recently described in mice and rats. In these proof-of-concept experiments, we show that intravenous injections of clinically relevant AAVs can transduce the ENS in guinea pigs and non-human primates. Neonatal guinea pigs were given intravenous injections of either AAV8 or AAV9 vectors that contained a green fluorescent protein (GFP) expression cassette or phosphate-buffered saline. Piglets were euthanized three weeks post injection and tissues were harvested for immunofluorescent analysis. GFP expression was detected in myenteric and submucosal neurons along the length of the gastrointestinal tract in AAV8 injected guinea pigs. GFP-positive neurons were found in dorsal motor nucleus of the vagus and dorsal root ganglia. Less transduction occurred in AAV9-treated tissues. Gastrointestinal tissues were analyzed from young cynomolgus macaques that received systemic injection of AAV9 GFP. GFP expression was detected in myenteric neurons of the stomach, small and large intestine. These data demonstrate that ENS gene delivery translates to larger species. This work develops tools for the field of neurogastroenterology to explore gut physiology and anatomy using emerging technologies such as optogenetics and gene editing. It also provides a basis to develop novel therapies for chronic gut disorders. Characterization of adeno-associated viral vector (AAV) mediated gene delivery to the enteric nervous system (ENS) was recently described in mice and rats. In these proof-of-concept experiments, we show that intravenous injections of clinically relevant AAVs can transduce the ENS in guinea pigs and non-human primates. Neonatal guinea pigs were given intravenous injections of either AAV8 or AAV9 vectors that contained a green fluorescent protein (GFP) expression cassette or phosphate-buffered saline. Piglets were euthanized three weeks post injection and tissues were harvested for immunofluorescent analysis. GFP expression was detected in myenteric and submucosal neurons along the length of the gastrointestinal tract in AAV8 injected guinea pigs. GFP-positive neurons were found in dorsal motor nucleus of the vagus and dorsal root ganglia. Less transduction occurred in AAV9-treated tissues. Gastrointestinal tissues were analyzed from young cynomolgus macaques that received systemic injection of AAV9 GFP. GFP expression was detected in myenteric neurons of the stomach, small and large intestine. These data demonstrate that ENS gene delivery translates to larger species. This work develops tools for the field of neurogastroenterology to explore gut physiology and anatomy using emerging technologies such as optogenetics and gene editing. It also provides a basis to develop novel therapies for chronic gut disorders. Gene Therapy (2017) 24, 640-648; doi: 10.1038/gt.2017.72; published online 7 September 2017 Characterization of adeno-associated viral vector (AAV) mediated gene delivery to the enteric nervous system (ENS) was recently described in mice and rats. In these proof-of-concept experiments, we show that intravenous injections of clinically relevant AAVs can transduce the ENS in guinea pigs and non-human primates. Neonatal guinea pigs were given intravenous injections of either AAV8 or AAV9 vectors that contained a green fluorescent protein (GFP) expression cassette or PBS. Piglets were euthanized three weeks post-injection and tissues were harvested for immunofluorescent analysis. GFP expression was detected in myenteric and submucosal neurons along the length of the gastrointestinal tract in AAV8 injected guinea pigs. GFP positive neurons were found in dorsal motor nucleus of the vagus and dorsal root ganglia. Less transduction occurred in AAV9 treated tissues. Gastrointestinal tissues were analyzed from young cynomolgus macaques that received systemic injection of AAV9 GFP. GFP expression was detected in myenteric neurons of the stomach, small and large intestine. These data demonstrate that ENS gene delivery translates to larger species. This work develops tools for the field of neurogastroenterology to explore gut physiology and anatomy using emerging technologies such as optogenetics and gene editing. It also provides a basis to develop novel therapies for chronic gut disorders.
Audience	Academic
Author	Fitzgerald, J A Todd, L J Mueller, C Kaspar, B K Cowley, C J Foust, K D Hook, K Lepak, C A Gombash, S E Bielefeld, E C Wood, J D Wang, G-D Neides, M G Fischer, A J
AuthorAffiliation	1 Department of Neuroscience, The Ohio State University, Columbus, OH, USA 2 SUCCESS Program, The Ohio State University, Columbus, OH, USA 5 Department of Pediatrics, The Ohio State University, Columbus, OH, USA 6 The Research Institute at Nationwide Children’s Hospital, Columbus, OH, USA 7 Department of Speech and Hearing Science, The Ohio State University, Columbus, OH, USA 3 Department of Physiology and Cell Biology, The Ohio State University, Columbus, OH, USA 4 Neurology & Gene Therapy Center, UMASS Medical School, Worcester, MA, USA
AuthorAffiliation_xml	– name: 4 Neurology & Gene Therapy Center, UMASS Medical School, Worcester, MA, USA – name: 6 The Research Institute at Nationwide Children’s Hospital, Columbus, OH, USA – name: 7 Department of Speech and Hearing Science, The Ohio State University, Columbus, OH, USA – name: 1 Department of Neuroscience, The Ohio State University, Columbus, OH, USA – name: 3 Department of Physiology and Cell Biology, The Ohio State University, Columbus, OH, USA – name: 5 Department of Pediatrics, The Ohio State University, Columbus, OH, USA – name: 2 SUCCESS Program, The Ohio State University, Columbus, OH, USA
Author_xml	– sequence: 1 givenname: S E surname: Gombash fullname: Gombash, S E email: sara.gombashlampe@osumc.edu organization: Department of Neuroscience, The Ohio State University – sequence: 2 givenname: C J surname: Cowley fullname: Cowley, C J organization: Department of Neuroscience, The Ohio State University – sequence: 3 givenname: J A surname: Fitzgerald fullname: Fitzgerald, J A organization: Department of Neuroscience, The Ohio State University – sequence: 4 givenname: C A surname: Lepak fullname: Lepak, C A organization: Department of Neuroscience, The Ohio State University – sequence: 5 givenname: M G surname: Neides fullname: Neides, M G organization: Department of Neuroscience, The Ohio State University – sequence: 6 givenname: K surname: Hook fullname: Hook, K organization: Department of Neuroscience, SUCCESS Program, The Ohio State University – sequence: 7 givenname: L J surname: Todd fullname: Todd, L J organization: Department of Neuroscience, The Ohio State University – sequence: 8 givenname: G-D surname: Wang fullname: Wang, G-D organization: Department of Physiology and Cell Biology, The Ohio State University – sequence: 9 givenname: C surname: Mueller fullname: Mueller, C organization: Department of Pediatrics, Neurology & Gene Therapy Center, UMASS Medical School – sequence: 10 givenname: B K surname: Kaspar fullname: Kaspar, B K organization: Department of Neuroscience, The Ohio State University, Department of Pediatrics, The Ohio State University, The Research Institute at Nationwide Children’s Hospital – sequence: 11 givenname: E C surname: Bielefeld fullname: Bielefeld, E C organization: Department of Speech and Hearing Science, The Ohio State University – sequence: 12 givenname: A J surname: Fischer fullname: Fischer, A J organization: Department of Neuroscience, The Ohio State University – sequence: 13 givenname: J D surname: Wood fullname: Wood, J D organization: Department of Physiology and Cell Biology, The Ohio State University – sequence: 14 givenname: K D surname: Foust fullname: Foust, K D organization: Department of Neuroscience, The Ohio State University
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Snippet	Characterization of adeno-associated viral vector (AAV) mediated gene delivery to the enteric nervous system (ENS) was recently described in mice and rats. In...
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Title	Systemic gene delivery transduces the enteric nervous system of guinea pigs and cynomolgus macaques
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