Multi-modality optical molecular imaging for melanoma tumor margin assessment

描述(申请人提供)：与任何其他类型的癌症相比，黑色素瘤的发病率和死亡率增长最快(5，6)。目前对所有阶段的黑色素瘤的治疗包括手术，通过肉眼检查和术后组织病理学评估原发灶的边缘。然而，对于消除局部复发和/或转移(3，12，13)所需的手术切缘的宽度仍然存在争议，这些情况对患者有严重的后果。此外，通过组织病理学实现的术后无肿瘤切缘评估有很大的局限性，包括1。 主观性，2.对切缘的监测只在切除的一小部分上进行，3.手术和手术成功的反馈之间的时间(即，确认切缘为负)可能需要数天时间。光学高分辨率3D分子成像提供了一种定量、非侵入性和实时评估边缘的方法。为此，这项建议侧重于基于定量分子和结构替代生物标记物的多模式光学方法来评估黑色素瘤肿瘤边缘。将使用两种分子成像方法。第一种是泵浦-探测显微镜，这是一种非线性光学方法，可以实现颜料的定量分子对比，包括真黑素和黄色素(12)。这种方法特别适用于黑色素瘤的诊断，因为黑素和黑色素在黑色素瘤的恶性进展中起着重要作用。第二种方法是泵浦-探测非线性相位色散光谱(PP-NLDS)，它通过探测交叉相位调制引起的相位变化来产生补充信息，这种普遍存在的效应提供了结构信息(35，36)，以及分子重定向效应(即光学克尔效应)，提供了对水环境(20，32)扩散性质的敏感性。其他结构信息将从多光子自发荧光、二次谐波产生和光学相干显微镜(OCM)中获得。该提案的第一个目标是开发一个综合的、多模式的系统。其战略是修改现有的定制显微镜，以纳入PP-NLDS和OCM，两者都是用相同的仪器检测到的。目标2将专注于利用人类移植皮肤小鼠模型制定适当的利润率评估标准。我们预计边缘呈阳性的病变将包含多种标记的组合，包括向真皮色素的转变、真皮微卫星、组织结构紊乱、细胞内环境的差异和细胞核增大。Aim 3将使用身体和接受手术的患者的人类样本来验证该方法。所有的成像结果将与组织病理学相关联，以验证该方法。这项工作的成功完成将为一种提供定量生物标记物来评估黑色素瘤肿瘤边缘的工具铺平道路，从而在不改变公认的临床范例的情况下，提高第一次手术成功的机会和及时。

DESCRIPTION (provided by applicant): Melanoma has the fastest growing incidence and mortality rates compared to any other type of cancer (5, 6). Current treatment for all stages of melanoma includes surgery with margins of the primary lesions assessed via visual inspection and post-operative histopathology. However, controversy still exists on the width of the surgical margins needed to eliminate local reoccurrence and/or in-transit metastases (3, 12, 13), conditions that have severe consequences for patients. In addition, post-operative assessment of tumor-free margins, achieved via histopathology, has significant limitations, including 1. It is subjective, 2. Surveillance of the margins is performed on just a small portion of the excision, and 3. The time between surgery and feedback on its success (i.e., confirmation of negative margins) can take days. Optical high-resolution, 3D, molecular imaging provides a means by which margins can be assessed quantitatively, non-invasively, and in real time. To this end, this proposal focuses on a multi-modality optical approach to assess melanoma tumor margins based on quantitative molecular and structural surrogate biomarkers. Two molecular imaging methods will be used. The first is pump-probe microscopy, a nonlinear optical method that achieves quantitative molecular contrast of pigments, including eumelanin and phoemelanin (12). This method is uniquely suited for diagnosing melanoma since eu- and pheo-melanin play an important role in the malignant progression of this disease (12). The second method, pump-probe nonlinear phase dispersion spectroscopy (PP-NLDS), yields complementary information by probing phase changes resulting from cross phase modulation, a ubiquitous effect that provides structural information (35, 36), and molecular reorientation effects (i.e., optical Kerr effect), which provides sensitivity to the diffusive properties of aqueous environments (20, 32). Additional structural information will be obtained from multiphoton autofluorescence, second harmonic generation, and optical coherence microscopy (OCM). The first aim of the proposal is to develop a combined, multi-modality system. The strategy is to modify an existing custom-built microscope to incorporate PP-NLDS and OCM, both detected with the same instrumentation. Aim 2 will focus on developing proper criteria for margin assessment using a human grafted skin mouse model. We expect that lesions with positive margins will contain a combination of markers including a shift toward eumlanic pigment, dermal microsatellite, disorganized tissue architecture, differences in intracellular environments, and enlarged cell nuclei. Aim 3 will validate the approach using human specimens from cadavers and patients undergoing surgery. All imaging results will be correlated with histopathology to validate the approach. Successful completion of this work will pave the way for an instrument that provides quantitative biomarkers to assess melanoma tumor margins, thereby improving the chances of surgical success in the first excision and in a timely manner, without altering accepted clinical paradigms.