Genetically-based neuro-modulation of adipose tissue functions

项目摘要 许多药物疗法利用自主神经系统的强大控制 (ANS)超过外周器官功能，但药物的意外副作用往往是一个 重要的问题。因此，更多的自主神经流出的功能特异性激活将是一个重要的因素。 是许多病症和疾病的治疗方案的一个受欢迎的补充。 在这里，我们建议应用神经元特异性操作的现代工具箱，例如 光遗传学和药物遗传学刺激和抑制ANS。该技术 基于携带抑制兴奋性或抑制性的指令的神经营养病毒载体 离子通道或受体进入特定的神经元群体，并已广泛用于中枢神经系统 啮齿类动物。由于该技术还没有在自动化系统中应用，我们将测试其可行性 不同的病毒和病毒构建体选择性地阻断交感神经（SNS）神经元 作为模型系统（目标1）支配脂肪组织，并使用基于基因的技术 生成在人体组织中验证的脂肪组织相关SNS神经元的基因谱。 这将产生i）具有经证实的选择性和有效地与细胞结合的能力的病毒构建体， 感染/切除节后SNS神经元，ii）化学特异性节后SNS图谱 神经元支配的棕色和白色脂肪组织的小鼠，和iii）地图的详细 脂肪组织垫的神经支配模式，用于将来的选择性去神经支配和电 刺激方法，iv）脂肪组织相关SNS神经元的遗传概况及其 在人类SNS神经元中的代表性。在目标2中，我们将测试急性和慢性 成功转染的节后SNS的光遗传学和药物遗传学刺激 支配棕色脂肪组织的神经元，以诱导产热、能量消耗和 体重减轻和支配皮下白色脂肪组织的SNS神经元诱导 脂肪分解和布朗宁。因此，我们将为基于基因的功能性 可以应用于任何其他节后自主神经元的特定ANS操纵， 如肝细胞和胰岛的去甲肾上腺素能神经支配，以及 整个胃肠道的肌间神经丛神经元的胆碱能（迷走）神经支配， 胰岛、胆道系统和肝脏。这将最终允许选择性地 操纵能量平衡和葡萄糖稳态的其他方面，以及 心血管、呼吸、泌尿生殖和胃肠道功能。

Project Summary Many pharmacotherapies capitalize on the powerful control of the autonomic nervous system (ANS) over peripheral organ functions, but unintended side effects of drugs are often an important issue. Therefore, more function-specific activation of autonomic outflow would be a welcome addition to the therapeutic repertoire for many conditions and diseases. Here we propose to apply the modern tool box of neuron-specific manipulation such as optogenetic and pharmacogenetic stimulation and inhibition to the ANS. This technology is based on neurotrophic viral vectors carrying instructions for transfecting excitatory or inhibitory ion channels or receptors into specific neuron populations and has been widely used in the CNS of rodents. Because the technology has not yet been used in the ANS, we will test the feasibility of different viruses and viral constructs to selectively transfect sympathetic (SNS) neurons innervating adipose tissue as a model system (Aim 1) and use genetically based technology to generate a gene profile of adipose tissue related SNS neurons that are verified in human tissue. This should result in i) viral constructs with proven ability to selectively and efficiently infect/transfect postganglionic SNS neurons, ii) maps of chemo-specific postganglionic SNS neurons innervating brown and white adipose tissue in the mouse, and iii) maps of detailed innervation patterns of adipose tissue pads for future selective denervation and electrical stimulation approaches, iv) genetic profile of adipose tissue related SNS neurons and their representation in human SNS neurons. In Aim 2, we will test the ability of acute and chronic optogenetic and pharmacogenetic stimulation of successfully transfected postganglionic SNS neurons innervating brown adipose tissue to induce thermogenesis, energy expenditure, and body weight loss and SNS neurons innervating subcutaneous white adipose tissue to induce lipolysis and browning. We will thus provide proof-of-principle for genetically-based functionally specific ANS manipulation that could be applied to any other postganglionic autonomic neuron, such as the noradrenergic innervation of hepatocytes and pancreatic islets, as well as cholinergic (vagal) innervation of myenteric plexus neurons throughout the gastrointestinal tract, pancreatic islets, the biliary system, and the liver. This will ultimately allow selective manipulation of other aspects of energy balance and glucose homeostasis as well as cardiovascular, respiratory, urogenital, and gastrointestinal functions in a modular fashion.