Closed Loop Wireless Monitoring and Optogenetic Modulation of Bladder Function

摘要 美国有数百万人患有膀胱功能障碍和间质性膀胱炎引起的疼痛。 膀胱炎/膀胱疼痛综合征和膀胱过度活动症。许多这些疾病的潜在病理 人们对此知之甚少，这是目前大多数治疗无效的主要原因。解决 本课题组设计并测试了一种植入式无线光电系统， 调节膀胱功能。我们的超适形应变片包裹在膀胱周围， 膨胀时，器件几何形状的变化线性地增加电阻，然后可以将电阻直接关联到 膀胱大小或充盈度。我们将微型发光二极管（μLED）连接到应变仪上， 可用于激活光敏视蛋白，用于神经元活性的光遗传学调节。应变仪 和μ LED连接到一个可植入的基站，允许无线控制和监测膀胱 活动这项新技术消除了植入潜在损害膀胱导管的需要， 电极，并提供了独特的访问膀胱功能在清醒，自由移动的大鼠。我打算利用 视蛋白的病毒递送和测量膀胱周长动态变化的新型应变仪， 分别调节和监测膀胱功能。我的初步数据显示应变仪的变化 阻力与传统的膀胱活动测量（如囊内压）相关， 递送的抑制性视蛋白，古视紫红质（Archaerhodopsin，Arch），可以延迟麻醉大鼠的膀胱收缩。我计划 为了测试在膀胱传入神经中表达的病毒转导的Arch降低频率和增加细胞增殖的能力， 清醒动物中环磷酰胺（CTX）诱导膀胱炎后的排尿量（目的1a和B）。我还计划 使用我们新开发的无线技术来实现一个闭环系统， 膀胱收缩的频率，并启动光遗传学抑制以使排尿正常化（目的1c）。膀胱疼痛 是IC/BPS患者最常见的主诉。使用我们的无线μLED应变仪，我计划 为了确定膀胱传入神经中病毒转导的Arch的激活是否足以 减轻CYP诱导的膀胱炎大鼠的膀胱超敏反应。该灵敏度将通过 内脏反应、腹部敏感性和真实的时间位置偏好测定（目的2）。执行 这项新技术将为了解膀胱功能提供独特的途径， 植入潜在损伤性膀胱导管或电极。因此，这项技术可能会导致新的 对膀胱控制和疼痛机制的见解。此外，这项新技术的改进 以及光遗传学通道的病毒递送方法，可能会导致未来患者治疗的发展 膀胱疼痛和功能障碍

ABSTRACT Millions of people in the United States suffer from bladder dysfunction and pain caused by interstitial cystitis/bladder pain syndrome and overactive bladder. The underling pathologies for many of these diseases are poorly understood and this is the primary reason why most current treatments are ineffective. To address this issue, our group has designed and tested an implantable wireless optoelectronic system to monitor and modulate bladder function. Our hyper-conformal strain gauge wraps around the bladder and as the bladder expands, changes in geometry of the device linearly increases resistance which can then be directly correlated with bladder size or fullness. We have attached microscale light emitting diodes (μLED) to the strain gauge that can be used to activate light-sensitive opsins for optogenetic regulation of neuronal activity. The strain gauge and μLEDs connect to an implantable base station that allows for wireless control and monitoring of bladder activity. This new technology eliminates the need for implantation of potentially-damaging bladder catheters or electrodes, and provides unique access to bladder functionality in the awake, freely-moving rat. I plan to utilize viral delivery of opsins and a novel strain gauge that measures dynamic changes in bladder circumference, to modulate and monitor bladder function, respectively. My preliminary data show that changes in strain gauge resistance correlates to traditional bladder activity measurements like intravesicular pressure, and that virally delivered inhibitory opsin, Archaerhodopsin (Arch), can delay bladder contractions in anesthetized rats. I plan to test the ability of virally transduced Arch expressed in bladder afferents to reduce frequency and increase voiding volume after cyclophosphamide (CYP) -induced cystitis in awake animals (Aim 1a and b). I also plan to use our newly developed wireless technology to implement a closed-loop system that can recognize increased frequency of bladder contractions and initiate optogenetic inhibition to normalize voiding (Aim 1c). Bladder pain is the most common complaint of patients suffering from IC/BPS. Using our wireless μLED strain gauge, I plan to determine whether activation of activation of virally transduced Arch, in bladder afferents, is sufficient to attenuate bladder hypersensitivity in rats with CYP-induced cystitis. This sensitivity will be assayed by visceromotor response, abdominal sensitivity and real time place preference assays (Aim 2). Implementation of this new technology will provide unique access to understanding bladder functionality without the need for implantation of potentially damaging bladder catheters or electrodes. This technology could thus lead to novel insights into the mechanisms of bladder control and pain. Additionally, the refinement of this novel technology and viral delivery methods for optogenetic channels, could lead to development of future therapies for patients with bladder pain and dysfunction.