An optogenetic-based control paradigm for neuromodulation of bladder function following spinal cord injury

项目摘要 每年有超过17，000名美国人患有脊髓损伤，其中许多患者患有 下尿路功能障碍这些膀胱疾病之一是逼尿肌-括约肌协同失调， 逼尿肌收缩以将尿液推出膀胱，尿道括约肌放松以 让尿液通过尿道，不能协调他们的行动，以产生一个空隙。电神经调制 技术可以有效地改善下尿路功能障碍的症状，但由于其复杂性， 涉及这些治疗的回路和神经系统不能启动协调的排尿收缩 点播为了解决这些局限性，我们计划开发一种光遗传学神经调节方法来靶向 并独立控制两个主要的神经系统（副交感神经和躯体运动）， 排尿反射光遗传神经调节涉及在神经细胞中遗传表达光学活化蛋白质。 神经元通过光刺激来调节它们的活动。使用这种方法，我们计划在大肠杆菌中表达兴奋性视蛋白。 通过使用启动子限制性表达和解剖学方法， 有限的病毒递送（目标1）。这些神经元的激活导致逼尿肌/膀胱收缩。抑制补药 尿道括约肌张力，我们将使用类似的方法来靶向支配 外尿道括约肌与抑制性视蛋白（目的2）。这些神经元的抑制导致括约肌松弛。 我们将测试表达的特异性、潜在的脱靶效应以及视蛋白的生理效应 在幼稚和脊髓损伤的大鼠模型中激活。最后，我们将开发一个控制范例， 协调逼尿肌激活和尿道括约肌松弛，以启动协调和完全排空 膀胱（目标3）。拟议的项目将开发创新的神经调节技术， 膀胱功能进一步开发精细和有效的神经调节技术是神经调节的第一步。 开发闭环疗法来治疗脊髓损伤相关的膀胱功能障碍。此外，本发明还 这些技术将在外周神经系统基因递送和细胞， 类型特异性神经调节。膀胱功能神经调节的进展将使我们更接近于 改善脊髓损伤患者的生活。

PROJECT SUMMARY More than 17,000 Americans suffer from spinal cord injuries every year, and many of these patients suffer from lower urinary tract dysfunction. One of these bladder disorders is detrusor-sphincter dyssynergia, where the detrusor muscle, which constricts to push urine out of the bladder, and the urethral sphincters, which relax to allow urine to pass out the urethra, cannot coordinate their actions to produce a void. Electrical neuromodulation technologies can be effective in improving symptoms of lower urinary tract dysfunction but due the complexity of the circuit and neural systems involved these therapies are unable to initiate a coordinated voiding contraction on-demand. To address these limitations, we plan to develop an optogenetic neuromodulatory approach to target and independently control the two main neuronal systems (parasympathetic and somatic motor) that are integral to the voiding reflex. Optogenetic neuromodulation involves genetically expressing optically activated proteins in neurons to modulate their activity with light stimulus. Using this method, we plan to express excitatory opsins in parasympathetic neurons innervating the detrusor by using promoter restricted expression and anatomically limited viral delivery (Aim 1). Activation of these neurons leads to detrusor/bladder contractions. To inhibit tonic urethral sphincter tone, we will use a similar approach to target the somatic motor neurons innervating the external urethral sphincter with inhibitory opsins (Aim 2). Inhibition of these neurons leads to sphincter relaxation. We will test the specificity of expression, potential off target effects, as well as the physiological effects of opsin activation in naïve and in a rat model of spinal cord injury. Finally, we will develop a control paradigm to coordinate activation of detrusor with urethral sphincter relaxation, to initiate coordinated and complete emptying of the bladder (Aim 3). The proposed project will develop innovative neuromodulatory techniques to control bladder function. Further development of refined and efficient neuromodulation techniques is the first step in developing closed-loop therapies to treat bladder dysfunction associated with spinal cord injury. Additionally, these techniques will have wide applicability in other fields of peripheral nervous system gene delivery and cell- type specific neuromodulation. Advances in neuromodulation of bladder function will bring us closer to goal of improving the lives of patients with spinal cord injury.