Fluorescence Spectroscopy to Detect Oral Neoplasia

描述（由申请人提供）： 口腔癌是世界范围内的一个主要健康问题。早期疾病的患者有更好的治愈机会和功能结果，但大多数患者存在晚期肿瘤。早期发现可改善口腔癌患者的预后。该提案的目标是开发一种新技术，荧光光谱法，用于非侵入性的，早期检测口腔肿瘤和评估与口腔癌变相关的分子变化。我们将检验这一假设，即在癌变过程中产生的生物化学和组织形态学的变化导致口腔粘膜的光学特性的改变。在目标1中，我们将根据使用荧光和反射光谱的临床试验结果开发诊断算法，并确定非侵入性识别和区分异型增生和早期癌与良性病变和正常粘膜的灵敏度和特异性。我们将在四个不同的源-检测器分离处获得荧光和反射光谱，这些分离从跨越上皮和浅表基质的不同深度采样信息。在目标2中，我们将使用人类口腔组织的短期培养来研究炎性和肿瘤性病变的荧光光谱变化的生物学基础。将正常和异常组织的宽视野横切片保存在培养物中，进行活体显微镜检查，以进行自体荧光显微镜检查。我们将比较正常、异型增生和癌变口腔粘膜的自发荧光模式;变化将与使用荧光光谱法在体内测量的变化相关。我们将通过比较自发荧光模式与荧光团的免疫组织化学模式（包括胶原交联、NADH、FAD、细胞角蛋白和卟啉）以及吸收和散射发色团（如血红蛋白），探索哪些发色团负责口腔粘膜荧光。有了这些信息，我们将评估荧光光谱作为癌症进展的中间终点生物标志物的潜力。在具体目标3中，我们将开发数学模型来描述口腔组织的荧光特性，以提取与肿瘤调制的主要发色团的相对贡献。我们将使用从该模型中提取的参数来开发直接基于组织生物化学和形态学变化的诊断算法，这些变化可以使用荧光进行探测。这项研究计划的成功完成将提供一个临床工具，可以大大提高早期发现和监测口腔肿瘤。非侵入性评估口腔粘膜分子变化的技术可以增强口腔肿瘤发生和治疗中涉及的遗传机制的临床和转化研究。

DESCRIPTION (provided by applicant): Oral cancer is a major health problem worldwide. Patients with early disease have better chances for cure and functional outcome, yet most patients present with advanced tumors. Early detection improves outcomes for oral cancer patients. The goal of this proposal is to develop a new technology, fluorescence spectroscopy, for non-invasive, early detection of oral cavity neoplasia and assessment of molecular changes associated with oral carcinogenesis. We will test the hypothesis that changes in biochemistry and tissue morphology produced during carcinogenesis result in alterations in the optical properties of oral mucosa. In Aim 1, we will develop diagnostic algorithms based on results of clinical trials using fluorescence and reflectance spectroscopy, and determine the sensitivity and specificity to non-invasively identify and distinguish dysplasia and early carcinoma from benign lesions and normal mucosa. We will obtain fluorescence and reflectance spectra at four different source-detector separations which sample information from different depths spanning the epithelium and superficial stroma. In Aim 2, we will investigate the biological basis for changes in fluorescence spectra of inflammatory and neoplastic lesions using short-term culture of human oral tissue. Vital microscopy using widefield Transverse slices of normal and abnormal tissue will be maintained in culture for examination with autofluorescence microscopy. We will compare the pattern of autofluorescence of normal, dysplastic and cancerous oral mucosa; changes will be related to those measured in vivo using fluorescence spectroscopy. We will explore which chromophores are responsible for oral mucosa fluorescence by comparing autofluorescence patterns to immunohistochemical patterns of fluorophores, including collagen crosslinks, NADH, FAD, cytokeratins and porphyrin, and absorbing and scattering chromophores such as hemoglobin. With this information, we will evaluate the potential of fluorescence spectroscopy as an intermediate endpoint biomarker of cancer progression. In Specific Aim 3, we will develop mathematical models to describe the fluorescence properties of oral tissue, to extract the relative contributions of principle chromophores modulated with neoplasia. We will use parameters extracted from this model to develop diagnostic algorithms based directly on alterations in tissue biochemistry and morphology that can be probed using fluorescence. Successful completion of this research program will provide a clinical tool that could dramatically improve early detection and monitoring of oral neoplasia. Technology to non-invasively assess molecular changes in oral mucosa could augment clinical and translational research of genetic mechanisms involved in carcinogenesis and treatment of oral neoplasia.