Fluorophore-Conjugated Antibodies for Imaging and Resection of GI Tumors

描述(由申请人提供)：我们建议加州大学圣地亚哥分校的研究人员(学术合作伙伴)和抗癌公司(工业合作伙伴)建立一个独特的学术-产业合作伙伴关系，以开发和验证荧光团结合抗体在手术导航和胃肠道肿瘤的腹腔镜定位中的使用。这项拟议的研究将为小鼠模型研究开发先进的成像技术、方法和工具，这些研究将可移植到临床，以开发荧光引导的癌症手术。假说抗肿瘤特异性抗原的荧光团标记抗体将改善原发和转移性胰腺癌和结肠癌的可视化、检测和切除。具体目的1利用荧光标记的针对肿瘤抗原CA19-9、CEA或两者的组合的单抗来促进胰腺癌和结肠癌的肿瘤边缘和转移灶的成像和切除。我们将使用抗肿瘤抗原CA19-9和CEA的荧光标记的单抗或两者的组合来评估在体人胰腺癌和结肠癌裸鼠原位转移模型中的肿瘤负荷，并促进原位和转移性病变的完全切除。将对荧光团结合的单抗进行毒性、剂量和组织分布研究。具体目的2我们将在我们的人胰腺癌小鼠模型中比较几种不同的荧光团在体内的剂量反应、体内信号持续时间、体内光漂白和体内信号背景比。荧光团在体内的初始荧光发射强度、荧光信号持续时间、被覆盖组织的散射和吸收以及随着长时间暴露在强光下而失去荧光强度的倾向有很大的不同，这一现象被称为光漂白。由于荧光团之间的这些差异，选择一个稳定的、波长和信号强度合适的荧光团用于体内是至关重要的。我们将在我们的人胰腺癌原位转移裸鼠模型中检测绿色(480-520 nm)、黄色(550-570 nm)、红色(610-650 nm)和远红(680-710 nm)范围内的几种不同的荧光团，以确定潜在的临床应用于人类癌症手术的最佳荧光团。具体目的3利用荧光腹腔镜术改善原发癌和转移性胃肠道癌的可视化，在正常的腹腔镜照明下是看不到的。通过腹腔镜术前清楚地区分所有荧光标记的癌症组织的能力，可以消除不必要的开腹手术的发病率，并指导胰腺癌和结肠癌的后续治疗。我们将使用由上述目标确定的抗体和荧光团的最佳组合来比较在荧光下的腹腔镜检查中与传统的腹腔镜检查中肿瘤检测的程度。 公共卫生相关性：结直肠癌和胰腺癌共同构成了美国癌症相关死亡的第三和第四大常见原因。对于这两种疾病，对原发和转移肿瘤的完整检测对患者的预后至关重要。我们的目标是利用日益增长的荧光成像技术，开发结直肠癌和胰腺癌术中分期的新方法，并提高我们在手术时实现真正切除的能力。

DESCRIPTION (provided by applicant): We propose a unique academic-industrial partnership between investigators at the University of California San Diego (Academic Partner) and AntiCancer, Inc. (Industrial Partner) to develop and validate the use of fluorophore-conjugated antibodies for surgical navigation and laparoscopic localization of gastrointestinal tumors. The proposed research will develop advanced imaging technology, methods and tools for mouse-model studies that will be translatable to the clinic to develop fluorescence-guided cancer surgery. Hypothesis Fluorophore-labeled antibodies against tumor-specific antigens will improve visualization, detection, and resection of primary and metastatic pancreatic and colon cancer. Specific Aim 1 Utilization of fluorophore-labeled monoclonal antibody specific for the tumor antigen CA19-9, CEA, or a combination of both to facilitate imaging and resection of tumor margins and metastatic lesions in pancreatic and colon cancer. We will use fluorescent-conjugated monoclonal antibodies against tumor antigen CA19-9 and CEA or a combination of both to evaluate tumor burden in vivo in an orthotopic metastatic nude mouse model of human pancreatic and colon cancer and to facilitate the complete resection of orthotopic and metastatic lesions. Toxicity, dosing, and tissue distribution studies will be performed for fluorophore-conjugated monoclonal antibodies. Specific Aim 2 We will compare several different fluorophores for in vivo dosing response, in vivo signal duration, in vivo photobleaching, and in vivo signal-to-background ratio in our mouse models of human pancreatic cancer. Fluorophores can vary greatly in their in vivo intensity of initial fluorescence emission, duration of fluorescence signal, scatter and absorption by overlying tissues and propensity for loss of fluorescence intensity with prolonged exposure to bright light, a phenomenon known as photobleaching. Due to these differences between fluorophores, it is of vital importance to choose a stable fluorophore with appropriate wavelength and signal intensity for in vivo use. We will examine several different fluorophores in the green (480-520 nm), yellow (550-570 nm), red (610-650 nm), and far-red (680-710 nm) ranges in our orthotopic metastastic nude mouse models of human pancreatic cancer to determine the optimal fluorophore candidates for potential clinical use for fluorescence guided cancer surgery in humans. Specific Aim 3 Utilization of fluorescence laparoscopy to improve visualization of primary and metastatic GI cancers not otherwise seen under normal laparoscopic lighting. The ability to clearly distinguish all fluorescently-labeled cancerous tissue pre-operatively via laparoscopy can eliminate the morbidity from an unnecessary laparotomy and direct subsequent treatment of pancreatic and colon cancer. We will use the optimal combination of antibodies and fluorophores as determined by the aims listed above to compare the extent of tumor detection during laparoscopy under fluorescence versus traditional laparoscopy. PUBLIC HEALTH RELEVANCE: Colorectal and pancreatic cancers together comprise the third and fourth most common causes of cancer- related death in the United States. For both diseases the complete detection of primary and metastatic tumor is critical to patient outcomes. Our goal is to utilize the growing technology of fluorescence imaging to both develop new methods of intraoperative staging for colorectal and pancreatic cancer and to improve our ability to achieve true resection at the time of surgery.