Dual orthogonal fluorescent protease sensors for image guided surgery

项目摘要 虽然治疗癌症的有效的化疗和放射学方法越来越多 治疗方面，主要的治疗策略仍然是手术切除肿瘤。最大的挑战 对于外科医生来说，1)准确和快速地定位原发病变，2)清楚地界定边缘 在肿瘤和正常组织之间。手术后以复发形式出现的失败通常是由于 不完全移除所有癌细胞。最近在光学材料的使用方面取得了一些进展 用于荧光成像引导手术的造影剂。然而，FDA批准的唯一一种光学 试剂是主要用于灌注成像的非靶向染料，在某些情况下会增加 肿瘤组织的摄取。一些分子靶向的光学造影剂最近已经发展到 临床试验。此外，还有一些光学探针可以利用酶的活性，例如 在肿瘤微环境中被激活的蛋白酶。我们已经开发出光学“智能探测器”， 只有当被激活的巨噬细胞中的一类蛋白酶作用时才会产生信号 肿瘤微环境。虽然这些探测器已经被证明是实时有价值的代理 在手术中使用FDA批准的达芬奇手术系统进行肿瘤成像，他们仍然遭受着相对 某些组织的背景较高，信号的动态范围有限，这取决于局部探针 浓度。为了解决这些限制，我们概述了一种范式转换的光学类的设计 需要多种肿瘤特异性酶信号才能激活的探针，以及FRET成像 提高探头的灵敏度和选择性的策略。我们将把这些探头与摄像机和 图像处理系统将进一步使他们最终的临床翻译成为可能。这项提议是建立在 围绕着大量的初步数据，这些数据来自博乔实验室和 Intuitive Surgical Inc.。这些初步结果验证了使用蛋白酶探针成像的关键概念 使用FDA批准的现有手术系统进行手术时的肿瘤边缘是一个强大的基础 为加强现有光学成像探头在外科手术中的应用而进行的拟议研究 工作流程。

Project Summary While there are an increasing number of useful chemotherapy and radiological methods available for cancer therapy, the predominant treatment strategy remains surgical resection of the tumor. The greatest challenges for the surgeon are 1) precisely and rapidly locating the primary lesion and 2) clearly defining the margins between tumor and normal tissues. Failure in the form of recurrence after surgery usually results from incomplete removal of all cancer cells. There have been a number of recent advances in the use of optical contrast agents for fluorescence imaging guided surgery (FIGS). However, the only FDA approved optical agents are non-targeted dyes that are mainly used for perfusion imaging and in some cases have increased uptake in tumor tissues. A number of molecularly targeted optical contrast agents have recently advanced to clinical trials. In addition, there are a number of optical probes that exploit the activity of enzymes such as proteases that are activated in the tumor microenvironment. We have developed optical ‘smart probes’ that produce a signal only when acted upon by a class of proteases found in activated macrophages within the tumor microenvironment. While these probes have already been proven to be valuable agents for real time tumor imaging during surgery using the FDA approved da Vinci surgical system, they still suffer from relatively high background in some tissues and limited dynamic range of signal that depends on local probe concentrations. To address these limitations, we outline the design of a paradigm-shifting class of optical probes that require multiple tumor-specific enzyme signatures for activation, as well as a FRET imaging strategy to enhance both sensitivity and selectivity of probes. We will couple these probes with camera and image processing systems that will further enable their eventual clinical translation. This proposal is built around a large body of preliminary data resulting from an active collaboration between the Bogyo Lab and Intuitive Surgical Inc. These initial results have validated the key concept of using protease probes to image tumor margins during surgery using existing FDA approved surgical systems and serve as a strong foundation for the proposed studies aimed at enhancing the utility of existing optical imaging probes in the surgical workflow.