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 双型鼻瘤肉瘤（SNS）是一个新近识别的肉瘤，影响女性三倍 经常比男人。这种高度侵入性病变的唯一治疗方法是毁容面部手术。我们最近 确定了SNS中的一种新颖的融合，该融合创建了一种新型的Chimera Pax3-MAML3，它融合了DNA结合 PAX3转录因子的域，具有策划者样3（MAML3）转录共激活因子。这 PAX3-MAML3触发的转录程序会引起恶性转化和侵入性增长，并且 SNS的性别二态性的基础是完全没有特征的。初步数据表明 PAX3-MAML3的表达足以驱动组织学模仿SN的肿瘤的形成，并 诱导SNS观察到的基因表达模式。值得注意的是，PAX3-MAML3诱导的雌激素表达 受体β（ERβ）和ERα的磷酸化，并刺激ER活性。此外，由 在女性患者中，异种移植的PAX3-MAML3表达细胞明显大于男性受体小鼠。 原代SNS肿瘤的转录组分析证实了体内雌激素反应签名的激活。 这些结果使我们假设ER在PAX3-MAML3的致病活性中起着至关重要的作用 在社交媒体中，这是该癌症的性别二态性的基础。我们进一步的海报，反 - 雌激素可能是一种完整的，甚至是毁灭手术的替代方法 治疗这种高度侵入性的癌症。这将通过以下测试：目标I：确定的作用 在体外和体内PAX3-MAML3发病机理中，我们将使用ER失活的影响，使用 在多个方面 PAX3-MAML3诱导的转化：（a）在体外，我们将监视对细胞增殖和存活的影响， 独立于锚固的生长和侵入性。 （b）在体内，我们将测试ER和雌激素的贡献 向PAX3-MAML3介导的肿瘤发生信号。我们将评估ER失活的影响（通过CRISPR- ERα/ERβ的介导的部署或用抗雌激素治疗小鼠，比较肿瘤生长 在雄性与雌鼠。 AIM II：确定PAX3-MAML3和ER坐标基因表达如何表达 驱动侵入性增长我们将：（a）定义由RNA-Seq pax3-MAML3引起的转录程序， 并确定如何通过ER调节（通过使用ER激动剂，以及CRISPR介导的删除 ERα/β）; （b）定义PAX3-MAML3和ER染色质占用范围以获得机械 了解PAX3-MAML3和ER如何协调此转录程序。转录组和芯片 - SEQ数据集将相交以区分直接与PAX3-MAML3和ER的间接靶基因，以及 洞悉对转录的合作影响。这种综合分析的长期目标是促进 识别PAX3-MAML3/ER目标及其下游途径，这对于 转化的，侵入性的表型，最终可能充当其他治疗靶标。 呢