Identification of Biomarkers for Testicular Cancer

描述（由申请人提供）： 睾丸生殖细胞肿瘤（TGCT）的发病率在过去40年中翻了一番。据报道，高加索人群的年增长率为3-6%。睾丸癌是影响14-40岁男性的最常见恶性肿瘤，占该年龄段诊断的所有恶性肿瘤的60%。尽管治愈率很高，但它们是该年龄组中实体瘤死亡的最常见原因。除了癌基因的激活，肿瘤抑制基因（TSGs）的失活已被证明在肿瘤发生中发挥重要作用。根据修正后的Knudson两击假说，肿瘤抑制基因（TSGs）的完全失活通常涉及两个遗传或表观遗传事件：1）含有一个亲本等位基因的大染色体DNA区域的丢失或重组和使第二个等位基因失活的局部突变事件2）一个等位基因中的局部突变和另一个等位基因中的甲基化3）一个等位基因中的洛缺失和另一个等位基因中的甲基化4）双等位基因甲基化我们最近发现了几个癌症特异性甲基化基因的一个强大的方法，夫妇概率搜索算法与药理学解蔽策略的无偏和精确的全球定位的肿瘤特异性甲基化基因。我们还验证了超过50个基因的高通量检测（QMSP，定量甲基化特异性PCR），用于以高灵敏度和特异性对几种类型的癌症进行稳健分析。与此同时，我们率先使用单核苷酸多态性（SNP）阵列分析癌症杂合性缺失（洛），并进行了证明的主要研究，使用基于SNP的阵列诊断膀胱癌的尿沉渣。我们现在提出以下目标：1。使用超过10，000个标记的单核苷酸多态性（SNP）阵列杂交分析不同阶段睾丸癌的染色体丢失模式。 2.通过一种稳健的方法发现睾丸癌特异性甲基化基因，该方法将整个人类基因组中的概率搜索算法与癌细胞系中已建立的药理学解蔽策略相结合。 3.比较不同阶段睾丸癌的基因组和表观基因组模式。在完成我们的前两个具体目标后，我们将确定涉及睾丸癌的遗传和表观遗传改变的关键区域中的一个或多个基因。此外，这些区域的识别以及遗传和表观遗传改变的比较将为睾丸癌进展的生物学提供重要的信息。功能性研究可以提供对生殖细胞永生化的第一步和随后的癌症进展的见解。最终，SNP标记和甲基化标记可能会导致睾丸癌的新诊断，监测和治疗方法。此外，这种全面的分析将使我们了解睾丸癌发展的生物学。

DESCRIPTION (provided by applicant): The incidence of testicular germ cell tumors (TGCT) has doubled in the past 40 years. An annual increase of 3-6% is reported for Caucasian populations. Testicular cancer is the most common malignancy affecting males aged 14-40 and accounting for up to 60% of all malignancies diagnosed at this age. Despite a high cure rate, they represent the most frequent cause of death from solid tumors in this age group. In addition to oncogene activation, the inactivation of tumor suppressor genes (TSGs) has been shown to play an important role in tumorigenesis. According to the revised Knudson's two-hit hypothesis, full inactivation of a tumor suppressor gene (TSGs) often involves two genetic or epigenetic events: 1) the loss or recombination of large chromosomal DNA regions containing one parental allele and a localized mutational event inactivating the second allele 2) localized mutation in one allele and methylation in another allele 3) LOH in one allele and methylation in another allele 4) biallelic methylation. We have recently discovered several cancer specific methylated genes by a robust approach that couples probabilistic search algorithms with the pharmacologic unmasking strategy for unbiased and precise global localization of tumor-specific methylated genes. We also validated a highthroughput assay for more than 50 genes (QMSP, quantitative methylation specific PCR) for robust analysis of several types of cancer with high sensitivity and specificity. In parallel, we have pioneered the use of single nucleotide polymorphism (SNP) arrays for analyzing cancer loss-of-heterozygosity (LOH) and performed a proof of principal study using SNP based arrays for the diagnosis of bladder cancer in urine sediment. We now propose the following aims: 1. To analyze patterns of chromosomal loss in testicular cancers from different stages using single nucleotide polymorphism (SNP) array hybridization of over 10,000 markers. 2. To discovery of testicular cancer specific methylated genes by a robust approach that couples probabilistic search algorithms in the entire human genome with an established pharmacologic unmasking strategy in cancer cell lines. 3. To compare the genomic and epigenomic patterns in different stages of testicular cancer. After carrying our first 2 specific aims, we will identify the gene or genes in a critical area of genetic and epigenetic alterations involved in testicular cancer. In addition, identification of these area and comparison of genetic and epigenetic alterations will shed important light on the biology of testicular cancer progression. Functional studies may provide insights into the first steps of germ cell immortilization and subsequent progression in cancer. Ultimately, SNP markers and methylation markers may lead to new diagnostic, monitoring and therapeutic approaches in testicular cancer. In addition this comprehensive analysis will lead us to understand the biology of testicular cancer development.