Preclinical development of a novel antibody conjugate for intraoperative detection of pancreatic cancer

项目摘要 胰腺癌未被诊断的原因有很多，包括解剖位置，模糊和非 局部症状，以及缺乏足够敏感和特异的疾病生物标志物。加上缺乏 尽管血液生物标志物中的大多数是非特异性的，但胰腺癌的早期检测仍然是一个重大挑战。可惜这 限制了最有效的治疗策略。因此，大约20%的患者 胰腺癌适合进行根治性手术。在这些患者中，大约75%的患者会复发， 在未来5年内，即使是那些被认为边缘阴性的患者也会出现这种疾病。尽管有着 胰腺癌患者的统计数据，现在有几种胰腺癌成像探头， 临床前和临床开发，以尝试改善疾病的去除。此外，增加使用 neoadjugant或诱导治疗导致更多的患者符合手术条件，如果他们 在一个较低的阶段重新登台。有机会开发针对胰腺癌的分子靶向探针， 手术和可能从手术中受益的患者数量的增加是关键的激励因素 项目粘蛋白-16（MUC 16）是胰腺癌的有吸引力的标志物，因为它在胰腺癌中高度过表达。 恶性肿瘤，但不是发炎或健康的胰腺。最近，我们评估了鼠抗MUC 16抗体， 称为AR9.6，与人MUC 16具有交叉反应性。在这些临床前研究中，我们证明了持久的 使用IRDy 800标记的AR9.6与使用AR9.6的胰腺癌增强相比， 同种型对照。使用胰腺癌异种移植物，AR9.6-IRDye 800能够有效地增强原位 和转移性疾病。转移检测是一个额外的好处，因为它可以防止患者 不必要的手术然而，鼠抗体不适合临床翻译。因此，A AR9.6的人源化形式现已由我们的合作团队开发。初步调查 与人源化AR9.6缀合的IRDye 800表明与MUC 16的强结合和体内靶向。因此，我们认为， 该项目的目标是进行NIRF标记的人源化AR9.6的临床前开发， 检测PDAC以改进R 0切除术并检测腹膜转移。这一目标将得到解决 通过三个具体目标：（1）开发和评估用于靶向MUC 16的人源化AR9.6-NIRF;（2）确定 人源化AR9.6-NIRF缀合物的临床前对比增强和安全性特征;和（3） 证明IRDye 800-AR9.6在胰腺癌临床前模型中的手术疗效。我们假设 胰腺癌中的异常MUC 16过表达将允许特异性靶向胰腺癌， 荧光标记的huAR9.6。这项研究项目的完成将导致荧光引导手术 造影剂能够（1）提高R 0胰腺癌切除率，以及（2）能够 识别腹膜转移，这将避免不必要的手术，并允许优化的治疗策略。

PROJECT SUMMARY Pancreatic cancer goes undiagnosed for a number of reasons including, anatomical location, vague and non- localizing symptoms, and a lack of adequately sensitive and specific biomarkers of disease. Combined with lack of blood biomarkers, early detection of pancreatic cancer remains a significant challenge. Unfortunately, this limits therapeutic strategies that are most efficacious. Consequently, approximately 20% of patients with pancreatic cancer are eligible for curative surgery. Of those patients, approximately 75% will have recurrent disease within the next 5 years, even those patients thought to have had negative margins. Despite the abysmal statistics for patients with pancreatic cancer, there are now several pancreatic cancer imaging probes in preclinical and clinical development to try to improve disease removal. Moreover, the increased use of neoadjugant or induction thereapy has resulted in additional patients that become eligible for surgery, if they restage at a lower stage. The opportunity to develop molecularly-targeted probes to pancreatic cancer to improve surgery and the increased numbers of patients that may benefit from surgery are key motivating factors for this project. Mucin-16 (MUC16) is an attractive marker for pancreatic cancer since it is highly overexpressed in malignancies, but not inflamed or healthy pancreas. Recently, we evaluated a murine antiMUC16 antibody, termed AR9.6, with cross-reactivity to human MUC16. In these preclinical studies, we demonstrated durable pancreatic cancer enhancement to at least 6 days postinjectin using IRDy800-labelled AR9.6 compared to an isotype control. Using a pancreatic cancer xenograft, AR9.6-IRDye800 was able to effectively enhance orthotopic and metastatic disease. Metastasis detection is an added benefit because it would prevent patients from unecessary surgery. Murine antibodies, however, are not suitable for clinical translation. Consequently, a humanized form of AR9.6 has now been developed by our collaborative team. An Initial investigation of IRDye800 conjugated to humanized AR9.6 suggests strong binding to MUC16 and in vivo targeting. Therefore, the goal of this project is to perform the preclinical development of NIRF-labelled, humanized AR9.6 to detect PDAC for improved R0 resection and to detect peritoneal metastasis. This goal will be addressed by three specific aims: To (1) develop and evaluate humanized AR9.6-NIRF for targeting of MUC16; (2) ascertain the preclinical contrast-enhancement and safety profile of humanized AR9.6-NIRF conjugates; and (3) demonstrate surgical efficacy of IRDye800-AR9.6 in preclinical models of pancreatic cancer. We hypothesize that the aberrant MUC16 overexpression in pancreatic cancer will allow specific targeting pancreatic cancer with fluorescently labelled huAR9.6. Completion of this research project will result in a fluorescence-guided surgery contrast agent that will be able to (1) improve the rate of R0 pancreatic cancer resections and (2) be able to identify peritoneal metastasis, which will prevent unnecessary surgery and allow optimized treatment strategy.