Pathogenic mechanisms of sinonasal sarcoma, a novel gender dimorphic cancer

一种新型性别二态性癌症——鼻窦肉瘤的发病机制

• 批准号: 10089419
• 负责人: Margaret Mary Chou
• 金额: $ 20.57万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10089419
• 关键词: Affect; Agonist; Anchorage-Independent Growth; Apoptotic; Area; Aromatase Inhibitors; Biological Assay; C-terminal; Carcinoma; Cell Proliferation; Cell Survival; Cells; ChIP-seq; Chimera organism; Chromatin; Clustered Regularly Interspaced Short Palindromic Repeats; DNA Binding Domain; Data; Data Set; Estrogen Antagonists; Estrogen Receptor alpha; Estrogen Receptor beta; Estrogens; Expression Profiling; FDA approved; Face; Female; Fibroblasts; Fulvestrant; Gender; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Goals; Growth; Histologic; Histology; Human; In Vitro; Incidence; Invasive Lesion; Lead; Malignant - descriptor; Malignant Neoplasms; Measures; Mediating; Mediator of activation protein; Messenger RNA; Molecular; Molecular Abnormality; Monitor; Morbidity - disease rate; Mus; Nature; Nose; Oncology; Operative Surgical Procedures; PAX3 gene; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Phenotype; Phosphorylation; Play; Recurrence; Reporter; Reporting; Role; Sampling; Signal Transduction; Tamoxifen; Testing; Time; Transcription Coactivator; Woman; Work; Xenograft procedure; base; cancer invasiveness; dimorphism; experience; gene induction; genome-wide; improved; in vivo; indexing; insight; leukemia; male; malignant breast neoplasm; men; middle age; mouse model; new therapeutic target; novel; programs; response; sarcoma; therapeutic target; transcription factor; transcriptome; transcriptome sequencing; tumor; tumor growth; tumorigenesis

PROJECT SUMMARY Bi-phenotypic sinonasal sarcoma (SNS) is a newly identified sarcoma that affects women three times more frequently than men. The only treatment for this highly invasive lesion is disfiguring facial surgery. We recently identified a novel fusion in SNS, which creates a novel chimera PAX3-MAML3 that fuses the DNA-binding domain of the PAX3 transcription factor with the Mastermind-like 3 (MAML3) transcriptional co-activator. The transcriptional program that PAX3-MAML3 triggers to elicit malignant transformation and invasive growth, and the basis of the gender dimorphism of SNS is completely uncharacterized. Preliminary data demonstrate that expression of PAX3-MAML3 is sufficient to drive formation of tumors that histologically mimic SNS, and to induce gene expression patterns observed in SNS. Notably, PAX3-MAML3 induced expression of estrogen receptor β (ERβ) and phosphorylation of ERα, and stimulated ER activity. Furthermore, tumors formed by xenografted PAX3-MAML3-expressing cells were significantly larger in female than male recipient mice. Transcriptome analysis of primary SNS tumors confirmed activation of an estrogen response signature in vivo. These results lead us to hypothesize that the ER plays an essential role in PAX3-MAML3's pathogenic activity in SNS, and that this underlies the gender dimorphic nature of this cancer. We further posit that anti- estrogens might serve as a complementary or even alternative approach to disfiguring surgery for the treatment of this highly invasive cancer. This will be tested through the following: Aim I: Determine role of ER in PAX3-MAML3 pathogenesis in vitro and in vivo We will examine the effects of ER inactivation, using CRISPR and anti-estrogenic agents (i.e. tamoxifen, fulvestrant, and aromatase inhibitor), on multiple aspects of PAX3-MAML3-induced transformation: (A) In vitro, we will monitor effects on cell proliferation and survival, anchorage-independent growth, and invasiveness. (B) In vivo, we will test the contribution of ER and estrogen signaling to PAX3-MAML3-mediated tumorigenesis. We will assess the effects of ER inactivation (by CRISPR- mediated depletion of ERα/ERβ or by treatment of mice with anti-estrogenic agents), comparing tumor growth in male vs. female mice. Aim II: Determine how PAX3-MAML3 and ER coordinate gene expression to drive invasive growth We will: (A) define the transcriptional program induced by PAX3-MAML3 by RNA-seq, and determine how it is modulated by ER (through use of ER agonists, and CRISPR-mediated deletion of ERα/β); and (B) define the PAX3-MAML3 and ER chromatin occupancy landscapes to gain a mechanistic understanding of how PAX3-MAML3 and ER coordinate this transcriptional program. Transcriptome and ChIP- seq data sets will be intersected to distinguish direct from indirect target genes of PAX3-MAML3 and ER, and to gain insight into cooperative effects on transcription. A longterm goal of this integrated analysis is to facilitate identification of PAX3-MAML3/ER targets and their downstream pathways that are essential for the transformed, invasive phenotype, and may ultimately serve as additional therapeutic targets. !

项目摘要 双表型鼻窦肉瘤是一种新发现的肉瘤，女性患者的发病率是男性的三倍 往往比男人。这种高度侵袭性病变的唯一治疗方法是面部毁容手术。我们最近 在SNS中发现了一种新的融合，它产生了一种新的嵌合体PAX 3-MAML 3， PAX 3转录因子的结构域与Mastermind样3（MAML 3）转录共激活因子。的 PAX 3-MAML 3触发以引发恶性转化和侵袭性生长的转录程序，和 SNS的性别二态性的基础是完全没有特征的。初步数据显示， PAX 3-MAML 3的表达足以驱动组织学上模拟SNS的肿瘤的形成， 在SNS中观察到诱导基因表达模式。值得注意的是，PAX 3-MAML 3诱导雌激素的表达， 受体β（ERβ）和ERα的磷酸化，并刺激ER活性。此外，由 异种移植的PAX 3-MAML 3表达细胞在雌性受体小鼠中显著大于雄性受体小鼠。 原发性SNS肿瘤的转录组分析证实了体内雌激素应答信号的激活。 这些结果使我们假设ER在PAX 3-MAML 3的致病活性中起重要作用 在SNS中，这是这种癌症的性别二态性的基础。我们进一步强调，反 雌激素可能作为一种补充，甚至替代方法，以毁容手术， 治疗这种高度侵袭性的癌症。这将通过以下方面加以检验： ER在体外和体内PAX 3-MAML 3发病机制中的作用 CRISPR和抗雌激素药物（即他莫昔芬、氟维司群和芳香酶抑制剂），在多个方面 PAX 3-MAML 3诱导的转化：（A）在体外，我们将监测对细胞增殖和存活的影响， 不依赖锚定的生长和侵袭性。(B)在体内，我们将测试ER和雌激素的贡献 信号传导至PAX 3-MAML 3介导的肿瘤发生。我们将评估ER失活的影响（通过CRISPR- 介导的ERα/ERβ耗竭或通过用抗雌激素剂治疗小鼠），比较肿瘤生长 在雄性和雌性小鼠中。目的II：确定PAX 3-MAML 3和ER如何协调基因表达， 我们将：（A）通过RNA-seq定义PAX 3-MAML 3诱导的转录程序， 并确定它是如何被ER调节的（通过使用ER激动剂，以及CRISPR介导的 ERα/β）;和（B）定义PAX 3-MAML 3和ER染色质占用景观，以获得对PAX 3-MAML 3和ER染色质占用景观的机制分析。 了解PAX 3-MAML 3和ER如何协调这一转录程序。转录组和ChIP- 将对seq数据集进行测序以区分PAX 3-MAML 3和ER的直接靶基因和间接靶基因，以及 以深入了解转录的协同效应。这一综合分析的长期目标是促进 确定PAX 3-MAML 3/ER靶点及其下游通路，这些靶点对于 转化的侵袭性表型，并可最终用作额外的治疗靶点。 !