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

项目总结 本申请建议将可激活的荧光素靶向胞浆磷脂酶A2α (cPLA2α)用于肺癌的术中检测。肺癌是导致癌症的主要原因。 死亡人数最多，是美国确诊人数第三多的癌症。非小细胞肺癌(NSCLC) 占肺癌的90%，手术切除仍然是最有效的治愈方法 低期转移前病变的患者。外科医生通常使用肉眼检查和手指触诊来 定义实体肿瘤边缘。然而，这种方法通常不足以检测残留病，从而导致 高达40%的患者复发，显著降低了5年生存率。 改善术中对肿瘤边缘的检测是当务之急，因为或 接近切缘是导致局部复发的最常见原因。最重要的预测 癌症手术后的指标是完全切除，这延长了患者的生存时间，并改善了术后 外科手术的生活质量。靶向近红外(NIR)成像用于肿瘤的对比增强 荧光体可以识别肉眼观察或触诊无法检测到的小病变。近红外成像 提供高分辨率和高灵敏度，并可在手术过程中实时执行。 在本申请中，我们建议翻译DDAO-花生四烯酸(DDAO-A)，这是一种旨在 特异性靶向cPLA2α，这是一种在肿瘤发生早期上调并过度表达的关键信号酶 在超过47%的非小细胞肺癌中。初步研究表明，DDAO-A可被小鼠和 在小鼠模型和切除的人肺肿瘤组织中都能检测到小病灶。 DDAO-A将在GLP条件下合成并分散在临床批准的脂质体配方中 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 检测肿瘤边缘的能力。系统毒性将在小鼠和一小群犬身上进行评估。我们 将利用已建立的近红外荧光团临床翻译平台，采用犬类临床试验 用于术中手术。我们将招募30只患有自发性非小细胞肺癌的同伴动物组成的患者队列 介绍给我们的兽医学校。 DDAO-A将用于术中检测原发肿瘤和确定肿瘤边缘 犬患者的荧光引导手术(FGS)。我们将研究全身注射和局部注射。 肿瘤残留检测申请表。这些数据将与对照手术队列进行比较 非靶向吲哚青绿(ICG)或无近红外荧光引导。如果成功，这项研究将 通过改善肺、淋巴和肺内小肿瘤沉积的定位来改善手术结果 利润率。这些研究的完成将为研究新药(IND)的申请奠定基础 以支持一项评估DDAO-A引导手术治疗人类肺癌患者的临床试验。

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.