Fluorophore-Conjugated Antibodies for Imaging and Resection of GI Tumors

描述（由申请人提供）：我们提议在加州大学圣地亚哥分校（学术合作伙伴）和抗癌公司（工业合作伙伴）的研究人员之间建立一个独特的学术-工业合作伙伴关系，以开发和验证荧光团偶联抗体在胃肠道肿瘤手术导航和腹腔镜定位中的使用。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，这些技术将转化为临床发展荧光引导的癌症手术。假设针对肿瘤特异性抗原的荧光标记抗体将改善原发性和转移性胰腺癌和结肠癌的可视化、检测和切除。特异性目的1利用针对肿瘤抗原CA19-9、CEA或两者结合的荧光标记单克隆抗体，促进胰腺癌和结肠癌肿瘤边缘和转移灶的成像和切除。我们将使用针对肿瘤抗原CA19-9和CEA的荧光偶联单克隆抗体或两者的组合来评估人胰腺癌和结肠癌原位转移裸鼠模型体内的肿瘤负荷，并促进原位和转移病变的完全切除。将对荧光基团偶联单克隆抗体进行毒性、给药和组织分布研究。我们将在人类胰腺癌小鼠模型中比较几种不同的荧光团的体内剂量反应、体内信号持续时间、体内光漂白和体内信号与背景比。荧光团在体内的初始荧光发射强度、荧光信号持续时间、上覆组织的散射和吸收以及长时间暴露在强光下荧光强度丧失的倾向（一种称为光漂白的现象）变化很大。由于荧光团之间存在这些差异，因此选择合适波长和信号强度的稳定荧光团在体内使用至关重要。我们将在我们的人类胰腺癌原位转移裸鼠模型中检查绿色（480-520 nm）、黄色（550-570 nm）、红色（610-650 nm）和远红色（680-710 nm）范围内的几种不同的荧光团，以确定用于荧光引导人类癌症手术的潜在临床应用的最佳荧光团候选者。利用荧光腹腔镜改善在正常腹腔镜照明下无法观察到的原发性和转移性胃肠道癌症的可视化。术前通过腹腔镜清晰区分所有荧光标记癌组织的能力可以消除不必要的剖腹手术和直接后续治疗胰腺癌和结肠癌的发病率。根据上述目标，我们将使用抗体和荧光团的最佳组合来比较腹腔镜下荧光与传统腹腔镜下肿瘤检测的程度。

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.