Quantitative multiphoton microscopy for non-invasive diagnosis of melanoma

 描述（由申请人提供）：摘要：多光子显微镜（MPM）是一种非线性激光扫描显微镜技术，具有高三维分辨率，快速成像能力和无标记分子对比度。几种内源性组织成分可以可视化，包括胶原蛋白（通过二次谐波产生，SHG），还原型烟酰胺腺嘌呤二核苷酸（NADH），黄素腺苷二核苷酸（FAD），角蛋白，黑色素和弹性蛋白纤维（通过双光子激发荧光，TPEF）。在不需要外源标记的情况下生成组织结构和组成的高分辨率图像的能力使得MPM成像特别适合于表征体内浅表组织。本临床成像提案的目的是评价体内多光子显微镜提供定量光学终点的能力，其具有足够高的预测能力，以可靠地区分三组色素性病变：常见痣、非典型痣和黑色素瘤。该框架是基于我们从15个病变（14例患者）研究中获得的初步结果，在该研究中，我们确定了三种与TPEF和SHG信号相关的光学生物标志物，并将这些体内分子特征与传统的离体组织病理学标准相关联。将MPM生物标志物组合以获得区分普通痣（MMI = 0-1）、非典型痣（MMI = 1-4）和黑色素瘤（MMI = 5-8）的定量9点数字指数（多光子黑色素瘤指数，MMI）（p<0.05）。我们现在提出了一个有把握的前瞻性临床试验，遵循这些有希望的结果，以确定是否MMI，或类似的MPM衍生指数，可以可靠地用于临床设置。我们希望我们的研究结果将提供一个有效的决策终点，以提高临床诊断的准确性，常见痣，非典型痣和黑色素瘤。此外，我们的努力，在体内MPM衍生的对比与传统的组织病理学，预计将导致新的见解的起源黑色素瘤的外观和进展。我们的长期目标是确定定量临床终点的正确组合，以改善临床诊断，指导有效治疗，并消除不必要的活检，同时增加需要切除的病变的识别。

 DESCRIPTION (provided by applicant): Summary: Multiphoton microscopy (MPM) is a nonlinear laser scanning microscopy technique that features high three-dimensional resolution, fast imaging capabilities and label-free molecular contrast. Several endogenous tissue components can be visualized, including collagen (through second-harmonic generation, SHG), reduced nicotinamide adenine dinucleotide (NADH), flavin adenosine dinucleotide (FAD), keratin, melanin and elastin fibers (through two-photon excited fluorescence, TPEF). The ability to generate high- resolution images of tissue structure and composition without the need for exogenous labels makes MPM imaging particularly well-suited for characterizing superficial tissues in vivo. The purpose of this clinical imaging proposal is to evaluate the ability of in viv multiphoton microscopy to provide quantitative optical endpoints with sufficiently high predictive power to reliably distinguish between pigmented lesions in three groups: common nevi, atypical nevi and melanoma. The framework is based on our preliminary results obtained from a 15-lesion (14 patient) study where we identified three optical biomarkers related to TPEF and SHG signals and correlated these in vivo molecular features with conventional ex vivo histopathologic criteria. MPM biomarkers were combined to obtain a quantitative, 9-point numerical index (multi-photon melanoma index, MMI) that distinguished between common nevi (MMI = 0-1), atypical nevi (MMI = 1-4) and melanoma (MMI = 5-8) (p<0.05). We now propose a powered prospective clinical trial that follows on these promising results in order to determine whether the MMI, or similar MPM-derived index, can be reliably used in a clinical setting. We expect our results will provide a validated decision-making endpoint to increase clinical diagnosis accuracy of common nevi, atypical nevi and melanoma. In addition, our effort to correlate in vivo MPM-derived contrast with conventional histopathology is expected to lead to new insight regarding the origins of melanoma appearance and progression. Our long-term goal is to identify the right combination of quantitative clinical endpoints that would improve clinical diagnoses, guide effective treatment, and eliminate unnecessary biopsies while increasing identification of lesions requiring removal.