RNA from Brush Cytology to Detect Squamous Cell Carcinoma

描述（由申请人提供）：鳞状细胞癌是一种累及包括胃肠道和气道粘膜在内的部位的癌症，是迄今为止口腔癌的主要形式。特别是，虽然口腔鳞状细胞癌（OSCC）通常以口腔中可见的病变开始，但在该可治愈阶段对疾病的早期检测是有限的。这可能是由于大量其他更无害的口腔病变的发生，可以类似于口腔鳞状细胞癌，以及手术活检的必要性，以准确检测和诊断疾病。口腔刷检细胞学可用于从可疑病变中收集细胞而无需手术，这反过来又可用于恶性变化的显微镜分析，但准确性降低。我们提出了一种病变评估的方法，使用这种相同的非侵入性程序，刷细胞学，但提供RNA，使病变粘膜的基因表达分析。我们已经证明，这种方法，相反的先入为主的想法，这将是不可行的，提供高质量的RNA能够允许从正常粘膜的仓鼠口腔癌模型中的口腔鳞癌的分化。我们将利用最近的研究，这些研究表明，在手术活检组织中，基于全局基因表达的变化，可以开发出能够准确诊断恶性疾病的分类预测因子。这项研究将确定是否可以分析来自细胞学的RNA，以产生可用于检测OSCC的全球基因表达特征。前瞻性OSCC患者，在接受标准活检诊断OSCC之前，将进行刷口腔细胞学检查，使我们能够开发，然后验证，识别OSCC的基因表达特征。一个直接的结果将是一个非侵入性的方法，以准确地检测和有助于口腔鳞状细胞癌的诊断，准备在其他诊所进一步验证。接下来，我们将应用同样的方法来开发一个基于基因表达的分类预测器，用于鳞状细胞癌的最早期、中度至重度异型增生和原位癌。在分析来自癌前口腔病变的刷检细胞学的RNA后，我们将使用针对异质性样本优化的统计方法，基于一小部分基因的基因表达来开发这些早期恶性肿瘤的类预测因子。最后，我们将开始确定使用刷检细胞学RNA检测和诊断口腔癌前病变和OSCC是否上级OralCDx系统。1 公共卫生相关性：这些结果应该为绝大多数口腔癌的检测提供一种非侵入性的方法，即使是在疾病的最早阶段也能得到验证。它将准备在其他诊所与其他研究者一起进行测试。这应该可以早期发现口腔癌，并可以立即挽救生命。这一发现将有力地支持来自刷检细胞学方法的RNA用于其他胃肠道和气道通道癌症。这项研究还将确定非侵入性刷检细胞学方法检测口腔癌前病变的可行性，并提供一个原型预测。通过一些修改，例如使用市售的室温RNA稳定溶液，这将使我们更接近一种经济，非侵入性的口腔癌筛查形式。

DESCRIPTION (provided by applicant): Squamous cell carcinoma, a cancer which afflicts sites including the gastrointestinal and airway mucosa, is by far the predominant form of oral cancer. Paradoxically, while oral squamous cell carcinoma, OSCC, typically initiates as visible lesions in the oral cavity, early detection of the disease at this curable stage is limited. This is likely due to the high number of other more innocuous oral lesions that occur that can resemble OSCC, and the necessity for a surgical biopsy for accurate detection and diagnosis of the disease. Brush oral cytology can be used to collect cells from suspect lesions without surgery, which in turn can be used for microscopic analysis of malignant changes but with decreased accuracy. We propose a method of lesion evaluation that uses this same noninvasive procedure, brush cytology, but to provide RNA to allow gene expression analysis of the lesion mucosa. We have shown that this methodology, contrary to preconceived ideas that it would not be feasible, provided high quality RNA capable of allowing the differentiation of OSCC from normal mucosa in a hamster oral cancer model. We will take advantage of recent studies that have shown in surgically biopsied tissue that class predictors, based on changes in global gene expression, can be developed that can accurately diagnose malignant disease. The study proposed will determine if RNA from cytology can be analyzed to produce a global gene expression signature that can be used to detect OSCC. Prospective OSCC patients, prior to undergoing standard biopsy diagnosis for OSCC, will be subjected to brush oral cytology to allow us to develop, and then validate, a gene expression signature that identifies OSCC. An immediate result will be a noninvasive methodology to accurately detect and contribute to the diagnosis of OSCC, ready to be further validated in other clinics. Next, we will apply this same approach to develop a gene expression based class predictor for the earliest stage of squamous cell carcinoma, moderate to severe dysplasia and carcinoma in situ. After analysis of RNA from brush cytology from pre-cancerous oral lesions, we will use statistical methodologies optimized for heterogeneous samples to develop a class predictor for these early malignancies based on gene expression of a small set of genes. In the end, we will start to determine if the detection and diagnosis of oral premalignancies and OSCC using RNA from brush cytology is superior to the OralCDx system. 1 PUBLIC HEALTH RELEVANCE: These results should deliver a noninvasive method for detection of the vast majority of oral cancers that is validated against even the earliest stages of the disease. It will be ready to be tested in additional clinics with other investigators. This should allow the earlier detection of oral cancer and could save lives immediately. This finding would strongly support the usage of the RNA from brush cytology approach with other gastrointestinal and airway passage cancers. This study would also determine the feasibility of noninvasive brush cytology methods to detect oral premalignancies and deliver a prototype predictor. With some modifications, such as the usage of commercially available room temperature RNA stabilization solutions, this would bring us much closer to an economical, noninvasive form of oral cancer screening.