Real-time Intraoperative Imaging of Cancer Biomarkers and Peripheral Nerves

项目概要/摘要： 黑色素瘤的早期诊断和治疗对于最大限度地减少其发病率和死亡率至关重要， 预后差，中位生存期不到1年。淋巴结转移的存在是至关重要的 因此，准确的识别对于在早期黑素瘤中准确分期黑素瘤是至关重要的。 阶段。前哨淋巴结（SLN）标测程序目前受到缺乏术中 可视化工具，可以帮助准确确定疾病扩散，并从相邻的 重要的神经和血管结构因此，更新的、变革性的和临床转化的外科手术 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， 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之外，并从相邻的正常神经中描绘出这些。超小型（小于10- nm）由黑素瘤靶向肽和PET放射性标记物修饰的荧光核-壳二氧化硅纳米颗粒 （124 I）代表一个这样的平台。一项基于早期αvβ3整合素的首次人体临床试验- 靶向颗粒探针康奈尔点（C点）显示出有利的药代动力学（PK）特征和体积 转移性黑色素瘤患者的肾排泄。扩展至自发性SLN标测研究 在黑色素瘤小型猪模型中，发现这种双模态整合素靶向颗粒特异性检测， 通过PET和图像引导定位和区分不同肿瘤负荷的荷癌淋巴结 术中近红外（NIR）荧光成像，后一种工具证实了光学活性结节 转移的直接可视化和病理相关性。为了提高和更准确地 在术中确定分子癌症表型，我们将利用这些整合素靶向 平台结合新开发的光谱独特的近红外CW 800染料含有C'和许多 更明亮的硅酸铝颗粒产品，AC'点携带肽靶向第二个公认的 癌症标志物，黑皮质素-1受体（MC 1-R），黑色素瘤细胞生长的促进剂。临床翻译 整合素靶向平台将用于为这些新一代MC 1类的开发提供信息， R瞄准C和AC点。本方案的目的是：（1）开发并验证一系列CW 800含染料 基于用于荧光标记的设计策略， 基于cRGDY的平台;（2）评估杂交124 I-αMSH-PEG-PEG的肿瘤选择性蓄积和PK特征。 表达MC 1-R的异种移植物、基因工程小鼠模型中的CW 800-C'点候选物，以及 黑色素瘤小型猪模型;（3）开发光谱不同的含NIR染料的产品， 同时区分病变淋巴结组织和正常组织类型上的不同病理标志物 用于术中光学成像引导;和（4）进行IND使能研究，用于临床试验开发。 这项工作的一个长期目标是通过识别 并鉴定可靠的生物标志物，这些生物标志物可以在我们未来的临床试验设计中得到验证，重点是 那些是癌症分期的

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.