Translating phospholipase activatable fluorophores for the sensitive detection of non-small cell lung cancer

项目摘要 本申请提出翻译靶向胞浆磷脂酶A2α的可激活荧光团 （cPLA 2 α）用于肺癌术中手术检测。肺癌是导致癌症的主要原因 是美国第三大确诊癌症。非小细胞肺癌（NSCLC）， 占肺癌的90%，手术切除仍然是最有效的治疗方法。 低阶段转移前疾病患者。外科医生通常使用目视检查和手指触诊来 定义实体瘤边缘。然而，这种方法通常不足以检测残留疾病，导致 高达40%的患者复发，5年生存率显著降低。 改进术中肿瘤边缘的检测是必要的，因为在手术中， 靠近切除边缘是局部复发的最常见原因。最重要的预后 癌症手术后的指标是完全切除，这提高了患者的生存率， 手术生活质量。使用靶向近红外（NIR）成像增强肿瘤的对比度 荧光团可以识别通过视觉观察或触诊无法检测到的小病变。NIR成像 提供高分辨率和灵敏度，并可在手术过程中实时执行。 在该应用中，我们提出翻译DDAO-花生四烯酸（DDAO-A），一种可激活的探针， 特异性靶向cPLA 2 α，一种在肿瘤发生早期上调并过表达的关键信号酶 超过47%的NSCLC。初步研究表明，DDAO-A被鼠和小鼠选择性激活， 人NSCLC肿瘤，并且可以检测小鼠模型和切除的人肺肿瘤组织中的小病灶。 DDAO-A将在GLP条件下合成，并分散在临床批准的脂质体制剂中 送货上门将进行选定的小鼠研究以确认探针激活、肿瘤靶向和肿瘤靶向。 检测肿瘤边缘的能力。将在小鼠和小型犬队列中评价全身毒性。我们 将利用已建立的近红外荧光团临床翻译平台，采用犬临床试验 用于术中手术。我们将招募30只患有自发性NSCLC的伴侣动物的患者队列 向我们的兽医学校介绍。 DDAO-A将用于术中检测原发性肿瘤和识别肿瘤边缘 荧光引导手术（FGS）在犬患者。我们将研究全身注射和局部 用于检测残留肿瘤。这些数据将与对照手术队列进行比较， 非靶向吲哚菁绿色（ICG）或没有NIR荧光引导。如果成功，这项研究将 通过改善肺、淋巴结和淋巴结中小肿瘤沉积物的定位来改善手术结果， 利润率这些研究的完成将构成研究性新药（IND）申请的基础 支持临床试验，以评估人类肺癌患者中DDAO-A引导的手术。

PROJECT SUMMARY This application proposes to translate activatable fluorophores targeted to cytosolic phospholipase A2α (cPLA2α) for intraoperative surgical detection in lung cancer. Lung cancer is the leading cancer-related cause of death and the third most diagnosed cancer in the United States. Non-small cell lung carcinomas (NSCLC), represent 90% of lung cancers, and surgical resection continues to be the most effective approach to cure patients with low stage premetastatic disease. Surgeons typically use visual inspection and finger palpation to define solid tumor margins. However, this approach is often insufficient to detect residual disease, leading to recurrence in up to 40% of patients and significantly reduced 5-year survival. Improving intraoperative detection of tumor margins is imperative, because small foci of residual disease at or close to the resection margins are the most common cause of local recurrence. The most important prognostic indicator following cancer surgery is complete resection, which prolongs patient survival and improves post- surgical quality of life. Contrast enhancement of tumors using near-infrared (NIR) imaging with targeted fluorophores can identify small lesions that are not detectable by visual observation or palpation. NIR imaging offers high resolution and sensitivity and can be performed in real-time during surgery. In this application, we propose to translate DDAO-arachidonate (DDAO-A), an activatable probe designed to specifically target cPLA2α, a critical signaling enzyme upregulated early in tumorigenesis and overexpressed in more than 47% of NSCLC. Preliminary Studies indicate that DDAO-A is selectively activated by murine and human NSCLC tumors, and can detect small foci in both mouse models and excised human lung tumor tissue. DDAO-A will be synthesized under GLP conditions and dispersed in clinically approved liposomal formulations for delivery. Selected mouse studies will be performed to confirm probe activation, tumor targeting and the ability to detect tumor margins. Systemic toxicity will be evaluated in mice and in a small cohort of canines. We will utilize an established platform for clinical translation of NIR fluorophores, employing a canine clinical trial for intraoperative surgery. We will recruit a patient cohort of 30 companion animals with spontaneous NSCLC presenting to our Veterinary School. DDAO-A will be used for detection of primary tumors and identification of tumor margins during intraoperative fluorescence-guided surgery (FGS) in canine patients. We will investigate both systemic injection and topical application for detection of residual tumor. These data will be compared to control surgical cohorts using the non-targeted indocyanine green (ICG) or without NIR fluorescence guidance. If successful, this study will improve surgical outcomes by improving localization of small tumor deposits in the lung, lymph nodes and margins. The completion of these studies will form the basis for an investigational new drug (IND) application in support of a clinical trial to assess DDAO-A-guided surgery in human lung cancer patients.