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

项目总结 胰腺癌被漏诊的原因有很多，包括，解剖位置，模糊和非 症状局部化，缺乏足够敏感和特异的疾病生物标志物。与缺乏相结合 在血液生物标志物中，胰腺癌的早期检测仍然是一个巨大的挑战。不幸的是，这 限制最有效的治疗策略。因此，大约20%的患者患有 胰腺癌有资格接受根治性手术。在这些患者中，大约75%的患者会复发 在接下来的5年内，即使是那些被认为边际利润为负的患者。尽管深不可测 对于胰腺癌患者的统计，现在有几种胰腺癌成像探头在 临床前和临床发展，试图改善疾病清除。此外，越来越多的人使用 新辅助或诱导治疗导致了更多有资格接受手术的患者，如果他们 在较低的阶段重新上演。开发针对胰腺癌的分子靶向探针的机会 手术和可能从手术中受益的患者数量的增加是导致这一现象的关键动机因素 项目。粘蛋白-16(MUC16)是胰腺癌的一个有吸引力的标志物，因为它在胰腺癌中高度过表达 是恶性肿瘤，但不是发炎或健康的胰腺。最近，我们评估了一种小鼠抗MUC16抗体， 命名为AR9.6，与人MUC16具有交叉反应。在这些临床前研究中，我们证明了耐受性 IRDy800标记的AR9.6在注射后至少6天对胰腺癌的增强作用 同型对照。使用胰腺癌异种移植瘤，AR9.6-IRDye800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，一个 AR9.6的人性化形式现在已经由我们的合作团队开发出来了。对……的初步调查 IRDye800与人源化AR9.6的结合表明与MUC16有很强的结合，并具有体内靶向性。因此， 该项目的目标是进行NIRF标记的人源化AR9.6的临床前开发，以 检测PDAC以改进R0切除，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)建立和评价针对MUC16靶向的人源化AR9.6-NIRF；(2)确定 人源化AR9.6-NIRF结合物的临床前对比增强和安全性； 在胰腺癌临床前模型中展示IRDye800-AR9.6的手术疗效。我们假设 胰腺癌中异常的MUC16过度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一种荧光引导手术 造影剂将能够(1)提高R0胰腺癌的切除率和(2)能够 识别腹膜转移，这将防止不必要的手术，并允许优化治疗策略。

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.