Project 1: Sex-specific developmental epigenetics in gliomagenesis

项目摘要 胶质母细胞瘤（GBM）是最具破坏性的脑癌形式。在接下来的一年里，大约22000人 美国人将患上GBM，几乎相同数量的人将死于GBM。而GBM既发生在男性身上， 我们可以可靠地预测，在22，000例新病例中，8，500例为女性，其余13，500例为女性。 病例为男性。此外，虽然明年女性GBM患者的中位生存期预计将是 大约22个月，男性则接近16个月。这些一致的分子基础 发病率和存活率的显著性别差异无法解释。在没有解释的情况下， 不可能完全知道在实验室中建模GBM和在实验室中治疗GBM的意义是什么。 诊所确定GBM性别差异的遗传和表观遗传机制 发病率和生存率是该项目的重点。我们最近发表了一项对GBM患者成像的分析， 转录组和生存率，我们确定女性GBM患者表现出更大的反应， 目前的标准治疗，并且它们的存活率与细胞因子的组分表达高度相关。 整合素信号通路。相比之下，男性GBM患者对当前治疗表现出较不稳健的反应， 它们的存活似乎更有效地由细胞周期调节机制的表达水平决定。 这些数据不仅为GBM生物学中的性别差异提供了新的见解，他们还表明性别特异性 针对支持女性和男性生存的途径可能会改善所有患者的结局。 由于性别差异，健康和疾病方面的性别差异在一生中不断累积。性 在受精时开始的分化，涉及遗传和表观遗传机制，以及 循环性激素的急性作用。我们已经建立了研究性别差异的小鼠模型 在GBM。在这里，我们将使用我们创新的Cas-9适应已建立的四核心基因型模型， 测量性染色体补体和性腺分泌物对性别差异的不同贡献 在GBM生物学上。再加上gRNA和其他遗传构建体的子宫内电穿孔，我们将是独一无二的。 定位于评估染色质结构和特定基因表达的性别特异性变化如何介导 GBM的性别差异。我们有两个目的来解决GBM发病率的性别差异这一假设， 和结果是在子宫内性分化的早期阶段确定的（目的1），并涉及性别特异性 整合素和细胞周期调控途径中基因表达和活性的模式化（Aim 2）。的各个阶段 这项工作，我们将纳入有关性别分化和铁代谢的具体问题（项目2）， 小胶质细胞功能（项目3）。这些研究将为我们提供关键信息， GBM性别差异的分子基础和实施性别特异性治疗的途径 GBM和其他癌症在发病率和结局方面表现出性别差异。

PROJECT SUMMARY Glioblastoma (GBM) is the most devastating form of brain cancer. In the next year, approximately 22,000 Americans will develop GBM and nearly the same number will die from it. While GBM occurs in both males and females, we can reliably predict that of the 22,000 new cases, 8,500 will be in females while the remaining 13,500 cases will be in males. Moreover, while the median survival for female GBM patients next year is expected to be approximately 22 months, for males it will be closer to 16 months. The molecular bases for these consistent and significant sex differences in incidence and survival are unexplained. In the absence of an explanation, it is impossible to fully know what the implications are for modeling GBM in the laboratory and for treating GBM in the clinic. Defining the genetic and epigenetic mechanisms that underlie sex differences in GBM incidence and survival is the focus of this project. We recently published an analysis of GBM patient imaging, transcriptomes, and survival in which we determined that female GBM patients exhibit greater response to the current standard treatments and that their survival is highly correlated with expression of components of the integrin signaling pathway. In contrast, male GBM patients exhibit less robust response to current treatment and their survival appears to be more potently determined by expression levels of the cell cycle regulatory machinery. These data not only provide new insights into sex differences in GBM biology, they suggest that sex-specific targeting of pathways that support survival in females and males could lead to improved outcomes for all patients. Sex differences in health and disease accrue throughout life as a consequence of sexual differentiation. Sexual differentiation, which begins at the time of fertilization, involves genetic and epigenetic mechanisms, as well as the acute actions of circulating sex hormones. We have developed murine models for studying sex differences in GBM. Here, we will use our innovative Cas-9 adaptation of the established four-core genotypes model for measuring the distinct contributions of sex chromosome complement and gonadal secretions to sex differences in GBM biology. Coupled with in utero electroporation of gRNAs and other genetic constructs, we will be uniquely positioned to assess how sex-specific changes in chromatin structure and expression of specific genes mediate the sex differences in GBM. We have two aims to address the hypothesis that sex differences in GBM incidence and outcome are determined at early stages of in utero sexual differentiation (Aim 1) and involve sex-specific patterning in gene expression and activity in integrin and cell cycle regulatory pathways (Aim 2). At all stages of this work we will incorporate specific questions about sexual differentiation and iron metabolism (Project 2) and microglia function (Project 3). Together these studies will provide critical information in our effort to understand the molecular basis for sex differences in GBM and a path for the implementation of sex-specific treatment for GBM and other cancers that exhibit sex differences in incidence and outcome.