Molecular Imaging Probe(s) for Optical Surgical Navigation of Pancreatic Cancer

摘要 胰腺癌是一种侵袭性的恶性肿瘤，手术切除是最有效的治疗方法 选项.对明亮的光可视化和触觉提示的依赖限制了PC的手术切除效率， 它的癌前病变，并有助于不利的结果。术中荧光引导 提高了肿瘤手术的分期和准确性，并降低了局部复发率 提高生存率。荧光引导成像的成功取决于灵敏度和 用于检测PC及其前驱病变的标记物的特异性。我们的多项研究 实验室和其他人已经确定了MUC 4在胰腺病理中的差异表达。虽然 在正常胰腺中检测不到，MUC 4的从头过表达仅限于高风险前体细胞， 病变和侵袭性PC，其表达逐渐增加疾病进展。一个内部 产生了针对MUC 4，8 G7的单克隆抗体（mAb），其识别串联中的重复表位 重复结构域，已成为一个有用的工具，超灵敏检测和确定MUC 4在肿瘤中的作用 进展和转移。PC中MUC 4的高表达与生存率低相关，而在前体细胞中MUC 4的高表达与生存率低相关。 MUC 4表达是恶性风险的预测因子。我们的初步研究表明， 施用的用NIRF染料IRDye 800 CW标记的mAb 8 G7可以非常灵敏地照射MUC 4表达细胞。 皮下和原位肿瘤。此外，我们还开发了一种独特的动物模型， 概括了MUC 4驱动的IPMN至侵袭性PC的进展。在该模型中，胰腺特异性诱导 人MUC 4与致癌KrasG 12 D的表达导致癌前病变的发展 进展为侵袭性PC的IPMN和PanIN病变。该提案旨在开发和评估MUC 4- 用于PC及其癌前病变临床前光学手术导航的靶向NIR探针 模型我们假设术中使用MUC 4靶向近红外荧光（NIRF） 成像探针将改善PC及其高危前驱病变的切除。为了验证这一 假设，提出了两个具体目标。目标1的研究集中在合成，表征和（预） MUC 4靶向近红外荧光缀合物的临床安全性分析。目的2研究评估 MUC 4靶向成像探针在患者来源的原位异种移植（PDOX）模型中的临床前功效， 人MUC 4转基因GEM模型和临床标本。总体而言，靶向成像的发展 探针可以改善前驱病变和PC的检测和无边缘切除。研究 在这项申请中提出的将开发和测试高性能的近红外探针靶向MUC 4（一个顶部 PC中差异过表达的膜粘蛋白）用于PC及其前驱病变中的手术导航。 研究目标的成功实现将为急需的I期临床试验铺平道路。 成像探头用于改善PC及其高危前驱病变的检测和手术切除。

ABSTRACT Pancreatic cancer (PC) is an aggressive malignancy where surgical resection is the most effective curative option. Reliance on bright light visualization and tactile cues limit the efficiency of surgical resection for PC and its premalignant precursor lesions and contribute to unfavorable outcomes. Intraoperative fluorescence guidance improves both stagings as well as the accuracy of oncologic surgeries and results in lower local recurrence rates and improved survival. The success of fluorescence-guided imaging is dependent on the sensitivity and specificity of a marker being utilized for detecting PC and its precursor lesions. Multiple studies from our laboratory and others have established the differential expression of MUC4 in pancreatic pathologies. While it is undetectable in the normal pancreas, de novo overexpression of MUC4 is restricted to high-risk-precursor lesions and invasive PC and its expression increases progressively disease advancement. An in-house generated monoclonal antibody (mAb) against MUC4, 8G7, which recognizes repetitive epitopes in the tandem repeat domain, has emerged as a useful tool for ultrasensitive detection and defining the role of MUC4 in tumor progression and metastasis. High MUC4 expression in PC is associated with poor survival, while in precursor lesions, MUC4 expression is a predictor of malignant risk. Our preliminary studies indicate that systemically administered mAb 8G7 labeled with NIRF dye IRDye800CW can very sensitively illuminate MUC4-expressing subcutaneous and orthotopic tumors in vivo. Further, we have developed a unique animal model that recapitulates MUC4-driven IPMN-to-invasive PC progression. In this model, pancreas-specific inducible expression of human MUC4 in conjunction with oncogenic KrasG12D results in the development of premalignant IPMN and PanIN lesions that progress to invasive PC. The proposal seeks to develop and evaluate MUC4- targeted NIR probes for optical surgical navigation of PC and its premalignant precursor lesions in the preclinical models. We hypothesize that intraoperative use of MUC4-targeted Near-Infrared Fluorescent (NIRF) imaging probes will improve the resection of PC and its high-risk precursor lesions. To test this hypothesis, two specific aims are proposed. Studies in Aim 1 focus on the synthesis, characterization, and (pre) clinical safety profiling of MUC4-targeted near-infrared fluorescent conjugates. Aim 2 studies evaluate the preclinical efficacy of MUC4-targeted imaging probes in patient-derived orthotopic xenograft (PDOX) models, human MUC4 transgenic GEM models, and clinical specimens. Overall, the development of targeted imaging probes can improve both the detection and margin-free resection of precursor lesions and PC. The studies proposed in this application will develop and test high-performance NIR probes targeting MUC4 (a top differentially overexpressed membrane mucin in PC) for surgical navigation in PC and its precursor lesions. Successful accomplishment of study goals will pave the path for a Phase I clinical trial for urgently needed imaging probes for improved detection and surgical resection of PC and its high-risk precursor lesions.