RNA from Brush Cytology to Detect Squamous Cell Carcinoma

描述（由申请人提供）：鳞状细胞癌是一种包括胃肠道和气道粘膜在内的部位的癌症，是目前口腔癌的主要形式。矛盾的是，虽然口腔鳞状细胞癌（OSCC）通常以可见的口腔病变开始，但在这个可治愈的阶段，这种疾病的早期发现是有限的。这可能是由于大量其他类似OSCC的无害口腔病变，以及需要进行手术活检以准确检测和诊断该疾病。刷式口腔细胞学可用于收集可疑病变的细胞，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。我们提出一种病变评估方法，使用相同的非侵入性程序，刷细胞学，但提供RNA以允许病变粘膜的基因表达分析。我们已经证明，与先入为主的不可行的想法相反，这种方法提供了高质量的RNA，能够使仓鼠口腔癌模型中的OSCC从正常粘膜分化。我们将利用最近的研究，这些研究表明，在手术活检组织中，基于整体基因表达的变化，可以开发出能够准确诊断恶性疾病的分类预测因子。提出的研究将确定是否可以分析细胞学中的RNA以产生可用于检测OSCC的全局基因表达特征。前瞻性OSCC患者，在接受标准的OSCC活检诊断之前，将接受口腔刷细胞学检查，以使我们能够开发并验证识别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，准确地检测和有助于OSCC的诊断，准备在其他诊所进一步验证。接下来，我们将采用相同的方法开发基于基因表达的分级预测器，用于早期鳞状细胞癌、中度至重度非典型增生和原位癌。在对癌前口腔病变的刷细胞学RNA进行分析后，我们将使用针对异质样本优化的统计方法，基于一小组基因的基因表达来开发这些早期恶性肿瘤的分类预测器。最后，我们将开始确定使用刷细胞学RNA检测和诊断口腔恶性前肿瘤和OSCC是否优于OralCDx系统。1

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.