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
Department of Neuroscience, The Ohio State University, Columbus, OH, USA.
Department of Neuroscience, SUCCESS Program, The Ohio State University, Columbus, OH, USA.
Gene Ther. 2017 Oct;24(10):640-648. doi: 10.1038/gt.2017.72. Epub 2017 Aug 3.
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.
最近在小鼠和大鼠中描述了腺相关病毒载体（AAV）介导的基因传递至肠神经系统（ENS）的特征。在这些概念验证实验中，我们表明静脉注射临床相关的AAV可转导豚鼠和非人类灵长类动物的ENS。给新生豚鼠静脉注射含有绿色荧光蛋白（GFP）表达盒的AAV8或AAV9载体或磷酸盐缓冲盐水。注射后三周对仔猪实施安乐死并采集组织进行免疫荧光分析。在注射AAV8的豚鼠胃肠道全长的肌间和黏膜下神经元中检测到GFP表达。在迷走神经背运动核和背根神经节中发现了GFP阳性神经元。在AAV9处理的组织中转导较少。对接受全身注射AAV9 GFP的幼年食蟹猕猴的胃肠道组织进行了分析。在胃、小肠和大肠的肌间神经元中检测到GFP表达。这些数据表明ENS基因传递适用于更大的物种。这项工作为神经胃肠病学领域开发了工具，以利用光遗传学和基因编辑等新兴技术探索肠道生理学和解剖学。它还为开发慢性肠道疾病的新疗法提供了基础。
