Fluorophore-Conjugated Antibodies for Imaging and Resection of GI Tumors

我们提出了一个独特的学术和工业之间的合作伙伴关系的研究人员在大学的 加州圣地亚哥（学术合作伙伴）和抗癌公司。（工业合作伙伴）开发和验证 荧光团偶联抗体在外科手术导航和腹腔镜定位中的应用 胃肠道肿瘤拟议的研究将开发先进的成像技术，方法和工具 用于小鼠模型研究，该研究可用于临床开发荧光引导的癌症手术。 假设针对肿瘤特异性抗原的荧光团标记抗体将改善可视化， 检测和切除原发性和转移性胰腺癌和结肠癌。 特异性目的1荧光团标记的肿瘤抗原特异性单克隆抗体的应用 CA 19 -9、CEA或两者的组合，以促进肿瘤边缘的成像和切除， 胰腺癌和结肠癌的转移性病变。我们将使用荧光标记的单克隆抗体 针对肿瘤抗原CA 19 -9和CEA的抗体或两者的组合，以评估在体内的肿瘤负荷。 人胰腺癌和结肠癌的原位转移性裸鼠模型，并促进 原位和转移病灶的完全切除。将进行毒性、给药和组织分布研究。 对于荧光团缀合的单克隆抗体进行。 具体目标2我们将比较几种不同的荧光团的体内给药反应， 信号持续时间，体内光漂白，和体内信号背景比在我们的小鼠模型， 人类胰腺癌荧光团在体内的初始荧光强度可以有很大的变化 荧光信号的发射、持续时间、上覆组织的散射和吸收以及 长时间暴露于强光下会导致荧光强度降低，这种现象称为光漂白。由于 针对荧光团之间的这些差异，选择稳定的荧光团是至关重要的， 适合于体内使用的波长和信号强度。我们将研究几种不同的荧光团， 在我们实验中，绿色（480-520 nm）、黄色（550-570 nm）、红色（610-650 nm）和远红色（680-710 nm）范围 人胰腺癌原位移植裸鼠模型，以确定最佳荧光团 用于荧光引导的人类癌症手术的潜在临床用途的候选者。 具体目标3利用荧光腹腔镜提高原发性和继发性肿瘤的可视化， 转移性胃肠道癌在正常的腹腔镜照明下看不到。能够清楚地 通过腹腔镜手术术前区分所有荧光标记的癌组织可以消除 不必要的剖腹手术和胰腺癌和结肠癌的直接后续治疗的发病率。 我们将使用由上述目标确定的抗体和荧光团的最佳组合， 比较荧光下腹腔镜检查与传统腹腔镜检查期间肿瘤检测的程度。

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.