Real-time Intraoperative Imaging of Cancer Biomarkers and Peripheral Nerves

项目摘要/摘要： 黑色素瘤的早期诊断和治疗对于将其发病率和死亡率降至最低至关重要 预后不良，中位生存期不足1年。淋巴转移的存在是至关重要的 因此，预后预测因素和准确的识别对于准确地对早期黑色素瘤进行分期是必不可少的。 阶段。前哨淋巴结(SLN)标测程序目前因术中缺乏而受到限制 可视化工具，可以帮助准确确定疾病传播并从相邻节点中勾勒出节点 关键的神经和血管结构。因此，更新、更具变革性和临床转译的外科手术 需要可视化工具来可靠地检测早期疾病，并提供更高的灵敏度和 多个癌症(和/或重要的正常组织)靶点的分子表征的准确性，包括 疾病扩散到SLN以外，并从邻近的正常神经勾画出来。超小型(低于10- 黑色素瘤靶向多肽和PET标记修饰的荧光核壳二氧化硅纳米粒子 (124I)代表一个这样的平台。基于早期一代αvβ3整合素的首个人类临床试验- 靶向粒子探针，康奈尔圆点(C点)，显示良好的药代动力学(PK)特征和块状 转移性黑色素瘤患者的肾脏排泄。扩展到自发性SLN标测研究 黑色素瘤迷你葡萄酒模型，这种双模整合素靶向颗粒被发现特异性地检测到， 用PET和图像引导定位和鉴别不同肿瘤负荷的荷瘤淋巴结 术中近红外(NIR)荧光成像，后者确定光学活跃结节 通过直接可视化和病理关联来判断转移情况。为了增强和更准确地 在术中环境中确定分子癌症表型，我们将利用这些整合素靶向 平台与新开发的光谱独特的近红外CW800染料C‘和MARE相结合 更亮的硅酸铝颗粒产品，AC‘圆点含肽靶向第二个成熟的 肿瘤标志物黑素皮质素-1受体(MC1-R)，黑色素瘤细胞生长的促进剂。临床翻译 整合素靶向平台将被用来通知这些新一代MC1- R瞄准C‘和AC’点。本方案的目的是：(1)开发和验证一系列含CW800染料 基于用于荧光的设计策略的针对MC1-R的黑色素瘤靶向颗粒 (2)评价杂合~(124)I-α的肿瘤选择性蓄积和PK谱。 CW800-C‘点在表达MC1-R的异种移植、基因工程小鼠模型和 黑色素瘤迷你葡萄酒模型；(3)开发光谱独特的含近红外染料的产品，允许 不同病理标记物在病变结节组织和正常组织类型中的同时鉴别 用于术中光学成像指导；以及(4)为临床试验开发执行启用IND的研究。 这项工作的一个较长期的目标是通过识别 以及鉴定可在我们未来的临床试验设计中验证的可靠生物标记物，重点是 那些用于癌症分期的。

Project Summary/Abstract: Early diagnosis and treatment of melanoma are essential to minimizing its morbidity and mortality given its very poor prognosis with median survival of less than 1 year. The presence of lymph node metastases is a vital prognostic predictor, and accurate identification is thus essential to accurately stage melanoma in its earlier phases. Sentinel lymph node (SLN) mapping procedures are currently limited by a lack of intraoperative visualization tools that can aid accurate determination of disease spread and delineate nodes from adjacent critical neural and vascular structures. Thus, newer, transformative and clinically translated surgical visualization tools are needed to reliably detect early-stage disease and offer improved sensitivity and accuracy in the molecular characterization of multiple cancer (and/or vital normal tissue) targets, including disease spread beyond the SLN, and the delineation of these from adjacent normal nerves. Ultrasmall (sub 10- nm) fluorescent core-shell silica nanoparticles modified by melanoma-targeting peptides and PET radiolabels (124I) represent one such platform. A first-in-human clinical trial based on an early generation αvβ3 integrin- targeting particle probe, Cornell dots (C dots), showed favorable pharmacokinetic (PK) signatures and bulk renal excretion in metastatic melanoma patients. Extended to SLN mapping studies in a spontaneous melanoma miniswine model, this dual-modality integrin-targeting particle was found to specifically detect, localize, and discriminate cancer-bearing lymph nodes of varying tumor burden by PET and image-directed intraoperative near-infrared (NIR) fluorescence imaging, the latter tool confirming optically-active nodal metastases by direct visualization and pathological correlation. In order to enhance and more accurately identify molecular cancer phenotypes in the intraoperative setting, we will utilize these integrin-targeting platforms in conjunction with newly developed and spectrally-distinct NIR CW800 dye-containing C' and much brighter alumina silicate particle products, AC' dots bearing peptides targeting a second well-established cancer marker, melanocortin-1 receptor (MC1-R), a promoter of melanoma cell growth. Clinically translated integrin-targeting platforms will be used to inform the development of these newer generation classes of MC1- R targeting C' and AC' dots. This proposal aims to: (1) develop and validate a series of CW800 dye-containing melanoma-targeting particles specific for MC1-R on the basis of design strategies utilized for fluorescent cRGDY-based platforms; (2) assess tumor-selective accumulations and PK profiles of hybrid 124I-αMSH-PEG- CW800-C' dot candidates in MC1-R-expressing xenografts, genetically engineered mouse models, and melanoma miniswine models; (3) develop spectrally-distinct NIR dye-containing products that allow for simultaneous differentiation of different pathological markers on diseased nodal tissue and normal tissue types for intraoperative optical imaging guidance; and (4) perform IND-enabling studies for clinical trial development. A longer-term objective of this work is to improve the efficiency of probe translation to the clinic by identifying and qualifying reliable biomarkers that can be validated in our future clinical trial designs with an emphasis on those for cancer staging.