Near-infrared Choline Kinase Sensors for Intraoperative Identification of Lung Tumor Margins

项目摘要 癌症手术后最重要的预后指标是完全切除。术中 肿瘤边缘的检测是具有挑战性的，残留疾病仍然是最常见的原因， 局部复发完全切除可延长患者生存期，改善术后质量， 生活使用具有靶向荧光团的近红外（NIR）成像对肿瘤进行视觉增强， 肉眼观察或触诊无法发现的病变。NIR成像提供高分辨率和 灵敏度，并且可以在术中环境中真实的时间执行。 本申请旨在测试靶向NIR荧光团用于肿瘤的鼠模型中的肿瘤检测的功效。 肺癌，并将这些发现用于指导犬患者的术中肿瘤切除术 患有自发性肺癌肺癌是导致死亡的主要癌症相关原因， 在美国确诊的癌症。在非小细胞肺癌（NSCLC）中，占90%， 临床肺癌，肿瘤切除术仍然是最有效的方法来治愈患者的局部 疾病外科医生通常使用目视检查和手指触诊来确定实体瘤边缘。 然而，这种方法往往不足以检测残留疾病，从而导致局部区域性的 高达40%的患者复发，并显著降低了该队列的5年生存率。 在这个提议中，我们将使用我们实验室合成的近红外探针来检测胆碱激酶 （ChoK），一种在肿瘤发生早期上调的脂质激酶，在56%的肺癌中过表达， 83%的NSCLC。这些探针将在两种NSCLC模型中检测其检测肿瘤边缘的能力， 小鼠我们将评估荧光团靶向和非靶向治疗后小鼠的肿瘤复发时间和存活率。 有针对性的手术切除从这些数据中，将选择一个荧光团用于GLP下的合成 条件和在犬队列中评价的全身毒性。然后我们将招募30名患者 患有自发性NSCLC的宾夕法尼亚大学兽医学院的伴侣动物。 我们的初步研究表明，犬非小细胞肺癌显著过表达ChoK，这种表达是由于 在整个肿瘤中是均匀的。我们将采用术中近红外荧光引导手术， 用于检测肿瘤和鉴定肿瘤边缘的ChoK探针。切除和恢复后，犬 将随时间监测患者的肿瘤复发和患者存活。这些数据将与 使用非靶向荧光团切除肿瘤的阳性和阴性对照组 吲哚花青绿色（ICG）或在没有NIR荧光引导的帮助下切除。这些研究将测试 我们的ChoK传感器检测犬自发性肺癌肿瘤边缘的能力 患者队列。我们的长期目标是开发用于真实的术中手术的靶向近红外荧光团 在人类癌症患者中。

PROJECT SUMMARY The most important prognostic indicator following cancer surgery is complete resection. Intraoperative detection of tumor margins is challenging, and residual disease continues to be the most common cause of local recurrence. Complete resection results in prolonged patient survival and improved post-surgical quality of life. Visual enhancement of tumors using near-infrared (NIR) imaging with targeted fluorophores can identify lesions that are not detectable by visual observation or palpation. NIR imaging offers high resolution and sensitivity and can be performed in real time in an intraoperative setting. This application aims to test the efficacy of targeted NIR fluorophores for tumor detection in murine models of lung cancer and to translate these findings to guide intraoperative surgical tumor removal in canine patients with spontaneous lung cancer. Lung cancer is the leading cancer-related cause of death and the third most diagnosed cancer in the United States. In non-small cell lung carcinomas (NSCLC), which represent 90% of clinical lung cancers, tumor resection continues to be the most effective approach to cure patients with local disease. Surgeons typically use visual inspection and finger palpation to define solid tumor margins. However, this approach is often insufficient for the detection of residual disease, thus leading to locoregional recurrence in up to 40% of patients and significantly reduced 5-year survival in this cohort. In this proposal, we will use NIR probes synthesized in our laboratory and designed to detect choline kinase (ChoK), a lipid kinase upregulated early in tumorigenesis and overexpressed in 56% of lung cancers and up to 83% of NSCLC. These probes will be tested for their ability to detect tumor margins in two models of NSCLC in mice. We will evaluate time to tumor recurrence and survival in mice after fluorophore-targeted and non- targeted surgical excision. From these data, one fluorophore will be selected for synthesis under GLP conditions and its systemic toxicity evaluated in a cohort of canines. We will then recruit a patient cohort of 30 companion animals presenting to the University of Pennsylvania Veterinary School with spontaneous NSCLC. Our Preliminary Studies indicate that canine NSCLC significantly overexpress ChoK and that this expression is homogeneous throughout the tumor. We will employ intraoperative NIR fluorescence-guided surgery using our ChoK probes for detection of tumors and identification of tumor margins. After resection and recovery, canine patients will be monitored over time for tumor recurrence and patient survival. These data will be compared to positive and negative control cohorts where tumors were resected using the non-targeted fluorophore indocyanine green (ICG) or resected without the aid of NIR fluorescence guidance. These studies will test the ability of our ChoK sensors to detect tumor margins in spontaneously occurring lung cancers in a canine patient cohort. Our long-term goal is to develop targeted NIR fluorophores for real time intraoperative surgery in human cancer patients.