Phenotype and Gene Expression in Odontogenic Tumors

描述（由申请人提供）：解开人类基因组导致我们对调节和决定正常和病理组织发育的分子遗传控制知识的快速进步。虽然在简单遗传条件方面取得了迅速和实质性的进展，但对复杂条件，包括导致和驱动肿瘤发生的分子事件的了解却少得多。分子生物学技术的发展使得同时分析大量基因成为可能，这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对正常和病理组织的表型和基因型之间的关系进行新的实验。先前的研究表明，RAS基因在牙源性肿瘤中过表达和/或突变，并在干细胞定位的位置发生肿瘤。我们假设，不同的牙源性肿瘤表型是由RAS和/或WNT通路下游的多个基因的差异表达引起的。我们建议通过确定具有不同表型病变的人的牙源性肿瘤来验证这一假设，这些病变包括成釉细胞瘤、牙瘤、牙源性角化囊肿和不常见的肿瘤类型。详细的表型档案将确定所有肿瘤使用临床，组织学和影像学技术。肿瘤间差异基因表达的表征将使用cDNA微阵列来完成，以评估16,000个小鼠或人类基因。肿瘤内特定细胞群的差异基因表达将使用激光捕获显微镜和微阵列进行评估。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因子。使用最先进的生物信息学技术对微阵列数据进行分析，以识别过度表达和表达不足的基因，并与商业上可用的参考RNA进行比较，从而允许实验室之间进行类似研究的标准化。基因数据将被及时地放在我们的网站上，以供数据共享。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.