Influence of Sex-Hormones in Meningiomas Pathogenesis

项目摘要 脑膜瘤患病率的显著女性偏倚，35-44岁患者的峰值比率超过3：1， 年龄和减少之后，很大程度上是无法解释的。已发表的临床证据强烈支持以下关系： 脑膜瘤和性激素;然而，机制尚不清楚。大多数激素受体表达研究 在最近脑膜瘤的遗传特征之前几十年进行的，现在已经被分类为 五个“基因组亚群”，按基因表达和解剖位置聚类。这些基因组亚群是 其特征在于在7个基因（AKT 1，KLF 4，POLR 2A，TRAF 7，PI 3 KCA， SMARCB 1和SMO），除了先前确定的最普遍的驱动因素之外，NF 2的损失， >80%的脑膜瘤。尽管所有的脑膜瘤都显示出女性的优势，但相关的临床数据超过2500例， 从我们的基因特征队列中发现，特别是两个亚组，KLF 4和POLR 2A， 独立地显示出在女性中不成比例的更大优势。了解亚组突变是否 对性激素的不同生理反应的结果可能为他莫昔芬失败的临床试验提供见解。 （II期）和米非司酮（III期），其显示出激动性或非显著的肿瘤反应性， 分别在脑膜瘤中。不幸的是，这两项试验都没有严格评估性激素受体或基因突变， status.我假设脑膜瘤驱动突变直接调节性激素信号网络， 基因表达的方式，有利于脑膜瘤的生长，通过一种机制，可预见的不同， 不同亚群的肿瘤。基于我的初步证据表明：（1）雌激素受体的表达 脑膜瘤中ER β和G蛋白偶联雌激素受体1蛋白与性激素表达的差异 受体蛋白跨越亚群;（2）人类脑膜对雌激素的双相生长反应，不分性别;（3） 激素和抗激素暴露后细胞活力以及雌激素反应元件反式激活的变化， 由复发性突变POLR 2AQ 403 K（占该基因中所有突变的> 85%）、KLF 4K 409 Q和 NF 2KO，如通过荧光素酶报告基因测定与正常脑膜和彼此比较所评估的;和（4）富集 通过增殖POLR 2AQ 403 K原代细胞来表达雌激素信号。据我所知，一项综合基因组研究 分类性激素受体转录激活和对染色质构象的影响，以前没有 在脑膜瘤中进行。我将通过两个目标来解决我的假设：具体目标1：调查关系 脑膜瘤基因组亚群与性激素受体状态之间的关系 （由Yale Pathology构建）并通过自动定量分析（ERα，ER β， GPER、AR和PGR）。具体目标2：阐明脑膜瘤基因组亚群突变是否调节性激素 受体转录活性通过进行ChIP-seq，RNA-seq，和基于酶的报告基因分析，对处理过的 含有各种脑膜瘤驱动突变的细胞。

PROJECT SUMMARY The remarkable female bias in the prevalence of meningiomas, with a peak ratio exceeding 3:1 in patients 35-44 years of age and decreasing thereafter, is largely unexplained. Published clinical evidence strongly supports a relationship between meningioma and sex hormones; however, the mechanism is unknown. The majority of hormone receptor expression studies were conducted decades before the recent genetic characterization of meningiomas, which have now been classified into five “genomic subgroups” that cluster by gene expression and anatomical location. These genomic subgroups are characterized by mutually exclusive somatic coding driver mutations in 7 genes (AKT1, KLF4, POLR2A, TRAF7, PI3KCA, SMARCB1, and SMO), which, in addition to the previously established most prevalent driver, the loss of NF2, account for >80% of all meningiomas. Although, all meningiomas show a female predominance, correlated clinical data of over 2,500 meningiomas from our genetically characterized cohort identified that two subgroups in particular, KLF4 and POLR2A, independently show a disproportionately greater predominance in females. Understanding whether the subgroup mutation results in distinct physiological response to sex hormones may provide insight into the failed clinical trials of tamoxifen (phase II) and mifepristone (phase III), which showed either an agonistic or non-significant tumor responsiveness, respectively, in meningiomas. Unfortunately, neither trial rigorously assessed sex-hormone receptor or genomic mutation status. I hypothesize that meningioma driver mutations directly modulate the sex-hormone signaling network to alter gene expression in a manner that is favorable to meningioma growth, by a mechanism that differs predictably between tumors of different subgroups. Based on my preliminary evidence demonstrating: (1) expression of the estrogen receptor  (ER) and G protein-coupled estrogen receptor (GPER1) proteins in meningiomas and variable expression of sex-hormone receptor proteins across subgroups; (2) biphasic growth response of human meninges to estrogen, regardless of sex; (3) changes in cell viability as well as estrogen-response element transactivation upon hormonal and anti-hormonal exposure, conferred by the recurrent mutations POLR2AQ403K (accounting for > 85% of all mutations in this gene), KLF4K409Q, and NF2KO, as assessed by luciferase reporter assay compared with normal meninges and with each other; and (4) enrichment of estrogen signaling by proliferating POLR2AQ403K primary cells. To my knowledge, an integrated genomic study classifying sex-hormone receptor transcriptional activation and influence on chromatin conformation has not previously been undertaken in meningiomas. I will address my hypothesis by two aims: Specific Aim 1: Investigate the relationship between meningioma genomic subgroups and sex-hormone receptor status by creating a Tumor Tissue Microarray (constructed by Yale Pathology) and assessing receptor protein expression by automated quantitative analysis (ERα, ER, GPER, AR, and PGR). Specific Aim 2: Elucidate whether meningioma genomic subgroup mutations modulate sex-hormone receptor transcriptional activity by conducting ChIP-seq, RNA-seq, and luciferase-based reporter assays on hormone-treated cells containing various meningioma driver mutations.