Novel Tools for Colon Cancer Detection and Therapy

结直肠癌(CRC)是美国的第三大癌症，占美国所有癌症的9.5%。 退伍军人中的癌症。结肠镜检查期间息肉切除术有显著意义 降低了结直肠癌发病率和相关患者死亡率。然而，尽管 虽然取得了重大进展，但减少CRC风险仍然是次优的，尤其是在 发生在右半结肠的结直肠癌。尽管内窥镜检查中的质量指标专注于 提高对标准腺瘤的检测，缺乏一致性 传统的腺瘤发现率和显示无梗锯齿状息肉的能力强调 需要辅助性方法。此外，尽管结直肠癌患者的R0切除率很高， 局部和远处复发仍然是一个严重的问题，被引用的比例高达40%。这个 肿瘤的完全切除对患者的预后至关重要。我们和其他人已经证明了 紧密连接蛋白CLDN-1在结直肠癌中的表达 在结肠癌的进展和发展过程中高度上调。 转移。多项研究进一步证实了Claudin-1结肠癌的致病作用 在体内和体外的进展和转移。简而言之，CLDN1的表达在一个阶段增加 特殊的方式和上调的CLDN1表达是癌前SSA/P的特征 腺瘤和结直肠癌转移。值得注意的是，在我们的研究中，9名转移性患者中有8人 原位异种移植(PDOX)显示CLDN1高表达。我们进一步展示了 利用抗体导向成像技术将抗CLDN1抗体与近红外荧光团清晰地结合 标记的肿瘤和肝转移实现了成功的荧光引导手术(FGS)。 据报道，CLDN1荧光肽对结直肠癌的检测能力类似 另一个实验室。综上所述，我们假设术中使用CLDN-1靶向近... 红外荧光(NIRF)成像探针将提高高危癌前病变的检测 腺瘤和结直肠癌转移，并提高其切除率。对于这一假设，我们提出了 以下具体研究：特定目标1：合成、表征和执行前 近红外荧光团标记的CLDN-1抗体的临床安全性评价。抗体染料 结合率提供最大的肿瘤信号，最小的短期和长期组织保留， 并将为临床前研究建立最低毒性。具体目标2：评估 CLDN-1靶向显像剂治疗息肉和结直肠癌的临床前和临床疗效 转移。CLDN-1-NIRF探针的临床指导效果也将在外科手术中得到验证 使用患者组织的体外成像从腺瘤和转移性肿瘤组织中提取样本。 对结直肠癌进展至关重要的关键分子的表征 开发新的早期发现和治疗方法不仅会减少患者 VA-CRC患者的死亡率，但也将有助于减轻相关的经济负担 退伍军人管理局。使用成像技术的临床前研究的结果 对于癌前息肉和结直肠癌的准确检测/切除将强烈影响 结直肠癌患者的管理。成功实现学习目标将为学习铺平道路 对于迫切需要的用于改进检测的新型成像探针的I期临床试验， 结直肠癌及其高危先兆病变的切除和处理。

Colorectal cancer (CRC) is the third major cancer in the USA and accounts for 9.5% of all the cancers among the Veterans. Polypectomy during screening colonoscopy has significantly reduced both the CRC incidence and associated patient mortality. However, despite the significant progress, the CRC risk reduction remains suboptimal and specifically worse in relation to CRC arising in the right colon. Although quality metrics in endoscopy have focused on improving detection of standard adenomas, there is a lack of concordance between the conventional adenoma detection rate and ability to visualize sessile serrated polyps underscoring the need for adjuvant approaches. Also, despite high R0 resection rates in patients with CRC, local and distant recurrence is still a significant problem and has been cited as high as 40%. The complete resection of tumor is critical to patient outcomes. We and others have shown that Claudin-1(CLDN-1), a tight junction (TJ) protein, expression increases in colorectal cancer (CRC) in stage specific manner, and is highly upregulated during colon cancer progression and metastasis. Several studies have further validated a causal role of Claudin-1 colon cancer progression and metastasis in vitro and in vivo. In brief, CLDN1 expression increases in a stage specific manner and an upregulated CLDN1 expression characterizes both, pre-malignant SSA/P adenomas and CRC metastasis. Notably, in our study, 8 out of 9 metastatic patients derived orthotopic xenografts (PDOX) demonstrated high CLDN1 expression. We further demonstrated that using antibody-guided imaging anti-CLDN1 antibodies conjugated to NIR fluorophores clearly labeled tumor and liver metastases enabling successful fluorescence-guided surgery (FGS). Similar ability of CLDN1 fluorescent peptides for detection of CRC adenomas was reported by another lab. Taken together, we hypothesize that intraoperative use of CLDN-1-targeted near- Infrared fluorescent (NIRF) imaging probes will improve the detection of high-risk pre-malignant adenomas and CRC metastasis, and also improve their resection. To this hypothesis, we propose following specific studies: SPECIFIC AIM 1: To synthesize, characterize, and perform the pre- clinical safety evaluation of CLDN-1 antibodies-conjugated to NIR fluorophores. The antibody-dye conjugate ratio providing maximum tumor signal, minimum short and long-term tissue retention, and lowest toxicity will be established for pre-clinical studies. SPECIFIC AIM 2: Assessment of the pre-clinical and clinical efficacy of CLDN-1-targeted imaging probes in polyps and CRC metastasis. The clinical-guiding efficacy of CLDN-1-NIRF probes will also be validated in surgical samples from adenomas and metastatic tumor tissues using ex vivo imaging of patient tissues. Characterization of the key molecules involved in the processes critical for CRC progression to develop novel early detection and therapeutic approaches would not only decrease patient mortality among VA-CRC patients but will also help reduce associated financial burden for the Veterans Administration. The outcome from the preclinical studies using the imaging technology for accurate detection/resection of premalignant polyps and CRC will strongly impact the management of CRC patients. Successful accomplishment of the study goals will pave the path for a Phase-I clinical trial for urgently needed novel imaging probes for improved detection, resection, and management of the CRC and its high-risk precursor lesions.