Influence of Sex-Hormones in Meningiomas Pathogenesis

项目总结 脑膜瘤患病率中明显的女性偏向，35-44岁脑膜瘤患者的峰值比率超过3：1 年龄和之后的下降，在很大程度上是无法解释的。已发表的临床证据有力地支持了 脑膜瘤和性激素；然而，其机制尚不清楚。大多数激素受体表达研究 是在脑膜瘤最近的基因特征之前几十年进行的，现在被归类为 根据基因表达和解剖位置聚在一起的五个“基因组亚群”。这些基因组亚群是 以7个基因(AKT1、KLF4、POLR2A、TRAF7、PI3KCA、 SMARCB1和SMO)，除了先前建立的最普遍的驱动因素外，NF2的丧失也占了 占所有脑膜瘤的80%。尽管所有脑膜瘤都以女性为主，但相关的临床数据超过2,500 来自我们的遗传特征队列的脑膜瘤确定了两个亚组，特别是KLF4和POLR2A， 在女性中独立表现出不成比例的更大优势。理解亚群突变是否 对性激素的不同生理反应结果可能为他莫昔芬失败的临床试验提供洞察 (II期)和米非司酮(III期)，这两种药物显示出激活性或无明显的肿瘤反应， 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组突变。 状态。我假设脑膜瘤驱动因素的突变直接调节性激素信号网络来改变 以一种有利于脑膜瘤生长的方式进行基因表达，其机制与可预测的 不同亚组的肿瘤。根据我的初步证据显示：(1)雌激素受体的表达 脑膜瘤中(ER)和G蛋白偶联雌激素受体1蛋白与性激素表达的变化 跨亚组的受体蛋白；(2)人类脑膜对雌激素的双相生长反应，不分性别；(3) 荷尔蒙和非荷尔蒙暴露后细胞活力以及雌激素反应元件反式激活的变化 由重复突变POLR2AQ403K(占该基因所有突变的85%)、KLF4K409Q和 用荧光素酶报告法测NF2KO，比较正常脑膜和彼此之间的差异；以及(4)浓缩 POLR2AQ403K原代细胞增殖对雌激素信号转导的影响。据我所知，一项完整的基因组研究 对性激素受体转录激活和对染色质构象的影响进行分类以前没有过 是在脑膜瘤中进行的。我将通过两个目标来解释我的假设：具体目标1：调查 脑膜瘤基因组亚群与性激素受体状态之间的关系 (由耶鲁病理学构建)，并通过自动定量分析(ERα，ER， GPER、AR和PGR)。特定目标2：阐明脑膜瘤基因组亚群突变是否调节性激素 激素处理后受体转录活性的ChIP-seq、rna-seq和荧光素酶报告分析 含有各种脑膜瘤驱动基因突变的细胞。

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.