Phenotype and Gene Expression in Odontogenic Tumors

描述（由申请人提供）：人类基因组的解开使我们对调节和决定正常和病理组织发育的分子遗传控制的认识迅速发展。虽然在简单遗传性疾病领域取得了快速和实质性的进展，但对复杂疾病（包括导致和驱动肿瘤发生的分子事件）的了解却少得多。同时分析大量基因的分子生物学技术的发展使得有效和高效地评估不同组织和/或肿瘤之间的基因表达成为可能。这些强大的分子技术允许新的实验到正常和病理组织的表型和基因型之间的关系。以往的研究表明，RAS基因的过度表达和/或突变的牙源性肿瘤和肿瘤的发生在干细胞定位的位置。我们推测，异质性牙源性肿瘤表型是由于RAS和/或WNT通路下游的多个基因的差异表达。我们建议通过确定具有表型多样性病变的人类牙源性肿瘤，包括成釉细胞瘤、牙瘤、牙源性角化囊肿和不太常见的肿瘤类型来验证这一假设。将使用临床、组织学和成像技术确定所有肿瘤的详细表型特征。肿瘤之间差异基因表达的表征将使用打印的cDNA微阵列来完成，以评估16，000个小鼠或人类基因。将使用激光捕获显微镜和微阵列评价肿瘤内特定细胞群的差异基因表达。将对RAS和p53基因进行突变测序，以发现肿瘤发生的分子启动子。使用最先进的生物信息学分析微阵列数据，以鉴定与市售参比RNA相比过度表达和表达不足的基因，从而允许进行类似研究的实验室之间的标准化。基因数据将及时放在我们的网站上进行数据共享。这些研究将首次提供人类牙源性肿瘤的详细突变和基因图谱研究。

DESCRIPTION (provided by applicant): Unraveling the human genome has lead to rapid advances in our knowledge of the molecular genetic controls that regulate and determine both normal and pathological tissue development. While progress has been rapid and substantial in the areas of simple hereditary conditions, far less has been learned of complex conditions including the molecular events leading to and driving tumorigenesis. The development of molecular biological techniques that allow analysis of large numbers of genes simultaneously now makes it possible to effectively and efficiently evaluate gene expression between different tissues and/or neoplasms. These powerful molecular techniques allow novel experimentation into the relationship between the phenotype and genotype of both normal and pathological tissues. Previous studies show overexpression of and/or mutation of RAS genes in odontogenic tumors and the genesis of tumors in locations where stem cells are localized. We hypothesize that the heterogeneous odontogenic tumor phenotypes result from differential expression of multiple genes that are downstream of the RAS and/or WNT pathways. We propose to test this hypotheses by ascertaining odontogenic tumors from humans having phenotypically diverse lesions including ameloblastomas, odontomas, odontogenic keratocysts and less common tumor types. Detailed phenotype profiles will be determined on all tumors using clinical, histological, and imaging techniques. Characterization of differential gene expression between tumors will be accomplished using cDNA microarrays printed to evaluate 16,000 mouse or human genes. Differential gene expression of specific cell populations within tumors will be evaluated using Laser Capture Microscopy and microarrays. RAS and p53 genes will be sequenced for mutations to discover molecular initiators of tumorigenesis. Microarray data is analyzed using state of the art bioinformatics to identify over and underexpressed genes compared with a commercially available reference RNA that allows standardization between laboratories conducting similar research. Gene data will be placed on our web site in a timely fashion for data sharing. These studies will provide the first detailed mutational and gene profile studies of human odontogenic tumors.