Genetically-based neuro-modulation of adipose tissue functions

项目摘要 许多药物疗法都利用对自主神经系统的强大控制。 (ANS)对周围器官功能的影响，但药物的意外副作用通常是 重要的问题。因此，更具功能特异性的自主神经流出激活将是一种 欢迎加入治疗多种疾病和疾病的保健品。 在这里，我们建议应用现代神经元特异性操作工具箱，如 光遗传和药物遗传对ANS的刺激和抑制。这项技术是 基于携带指令的神经营养病毒载体，用于转染兴奋性或抑制性 离子通道或受体进入特定的神经元群体，并已广泛应用于中枢神经系统 啮齿类动物。由于该技术尚未在自动化系统中使用，我们将对其可行性进行测试。 用不同的病毒和病毒构建物选择性地转染交感神经元 将脂肪组织作为模型系统(目标1)并使用基于基因的技术来 生成与脂肪组织相关的SNS神经元的基因图谱，并在人体组织中得到验证。 这将导致i)病毒构建体具有被证实的选择性和高效地 感染/转染节后SNs神经元，II)化学特异性节后SNs的图谱 支配小鼠棕色和白色脂肪组织的神经元，以及III)详细的 脂肪组织垫未来选择性去神经和电的神经支配模式 刺激途径，IV)脂肪组织相关三叉神经节神经元的遗传图谱及其 在人类三叉神经节神经元中的表达。在目标2中，我们将测试急性和慢性的能力 成功转染节后神经元的光遗传和药物遗传刺激 支配棕色脂肪组织的神经元诱导产热、能量消耗和 体重减轻与支配皮下白色脂肪组织的SNS神经元诱导 脂肪分解和褐变。因此，我们将为基于基因的功能提供原则证明 可以应用于任何其他节后自主神经元的特定ANS操作， 如肝细胞和胰岛的去甲肾上腺素能神经支配，以及 胃肠道肌间神经丛神经元的胆碱能(迷走神经)神经支配， 胰岛、胆道系统和肝脏。这最终将允许选择性地 控制能量平衡和葡萄糖动态平衡的其他方面以及 心血管、呼吸、泌尿生殖和胃肠功能模块化。

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.