NIR Nerve-specific Fluorescent Imaging for Robotic-assisted Radical Prostatectomy

项目摘要 手术后神经损伤是美国每年≥ 600，000例患者的持续发病率 一个人目前还没有临床上批准的方法来增强手术室中的直接神经可视化。 保留神经的脊髓切除术是一个令人信服的临床例子， 据报告，≥60%的患者出现损伤，导致失禁和阳痿。发展近 在手术室中专门突出神经组织的红外（NIR）荧光团将具有直接的 通过FDA批准的荧光通道，临床转化为保留神经的脊髓切除术， 芬奇手术机器人系统（Intuitive Surgical），目前用于≥80%的机器人辅助神经 保留根治性脑切除术（RARP），并将用于本文的猪成像。我们实验室以前的工作 神经特异性荧光团的合成库，当施用时， 系统地。然而，前列腺结构受到高度神经支配，并且所有神经结构都有系统标记 与直接标记包含负责神经的神经血管束相比， 功能。直接荧光团标记方法将在本文中验证，用于体内神经成像。 啮齿动物和猪模型，其中神经信号与背景比（SBR）的生成等同于或优于 将实现系统管理。前列腺的神经成像提出了一个额外的挑战， 器官被脂肪组织包围。我们已经表明，荧光团的理化性质，在 穿过血神经屏障（BNB）的范围也经常表现出亲脂性和脂肪积聚， 系统管理。我们已经确定，共同管理一个光谱不同，非特异性 荧光团可用于区分特异性和非特异性组织摄取。这项战略将 研究，在那里我们将检查光谱不同的近红外脂肪和神经特异性的共同管理。 与施用神经特异性荧光团相比，它们增强神经SBR的能力 单独的荧光团。在拟议的工作中，我们将合成一个以我们的铅为模型的聚焦恶嗪库， 神经特异性红移恶嗪4荧光团。400个合成的恶嗪荧光团将被 使用我们先前开发的测定法表征光谱性质和组织生物分布。两 将使用恶嗪4作为模型在啮齿动物和猪神经中研究直接给药策略 神经特异性荧光团，以确定提供最高神经SBR的策略。双重施用 光谱不同的恶嗪神经和脂肪特异性荧光团将被测试，以提高神经SBR 相对于单独给予神经特异性荧光团。NIR恶嗪荧光团将被配制用于体内 研究，其中优化的直接给药策略将用于啮齿动物。定量啮齿动物神经 SBR将用于选择配制的NIR恶嗪，以扩展到猪研究，该研究将在 与Intuitive Surgical合作，在da芬奇手术机器人中使用荧光通道。

PROJECT SUMMARY Nerve damage following surgery is a continued morbidity experienced by ≥600,000 patients annually in the US alone. Currently no clinically approved method exists to enhance direct nerve visualization in the surgical suite. Nerve-sparing prostatectomy is a compelling clinical example of significant patient morbidity, where nerve damage is reported in ≥60% of patients resulting in incontinence and impotence. Development of a near infrared (NIR) fluorophore that specifically highlights nerve tissue in the operating room would have direct clinical translation to nerve sparing prostatectomy through the FDA approved fluorescence channel in the da Vinci surgical robotic system (Intuitive Surgical), which is currently utilized for ≥80% of robotic assisted nerve sparing radical prostatectomies (RARP) and will be used for swine imaging herein. Previous work from our lab synthesized libraries of nerve-specific fluorophores that bind to all nerve tissue when administered systemically. However, the prostate structure is highly innervated and systemic labeling of all nerve structures is less desirable compared to directly labeling the neurovascular bundle that contains the nerves responsible for function. A direct fluorophore labeling methodology will be validated herein for in vivo nerve imaging in rodent and swine models, where generation of nerve signal to background ratio (SBR) equivalent or better than systemic administration will be achieved. Nerve imaging in the prostate presents an added challenge as the organ is surrounded by adipose tissue. We have shown that fluorophores with physiochemical properties in the range to cross the blood nerve barrier (BNB) also often exhibit lipophilicity and adipose accumulation upon systemic administration. We have established that co-administration of a spectrally distinct, non-specific fluorophore can be used to differentiate specific and non-specific tissue uptake. This strategy will be investigated, where we will examine the co-administration of spectrally distinct NIR adipose- and nerve-specific fluorophores for their ability to enhance nerve SBR as compared to administration of a nerve-specific fluorophore alone. In the proposed work, we will synthesize a focused oxazine library modeled after our lead, nerve-specific, red-shifted Oxazine 4 fluorophore. The 400 synthesized oxazine fluorophores will be characterized for spectral properties and tissue biodistribution using our previously developed assays. Two direct administration strategies will be investigated in rodent and swine nerves using Oxazine 4 as a model nerve-specific fluorophore to determine the strategy that provides the highest nerve SBR. Dual administration of spectrally distinct oxazine nerve- and adipose-specific fluorophores will be tested to enhance nerve SBR over nerve-specific fluorophore administration alone. NIR oxazine fluorophores will be formulated for in vivo studies, where the optimized direct administration strategy will be used in rodents. Quantified rodent nerve SBR will be used to select the formulated NIR oxazines to scale to swine studies, which will be conducted in collaboration with Intuitive Surgical using the fluorescence channel in a da Vinci surgical robot.