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

基本信息

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
项目类别：
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

项目摘要

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. !

项目概要双表型鼻窦肉瘤 (SNS) 是一种新发现的肉瘤，对女性的影响是女性的三倍这种高度侵入性病变的唯一治疗方法是毁容面部手术。在 SNS 中发现了一种新的融合，它创造了一种新的嵌合体 PAX3-MAML3，它融合了 DNA 结合 PAX3 转录因子的结构域与 Mastermind-like 3 (MAML3) 转录共激活因子。 PAX3-MAML3 触发的转录程序引发恶性转化和侵袭性生长，以及初步数据表明，SNS 性别二态性的基础完全未知。 PAX3-MAML3 的表达足以驱动组织学上模仿 SNS 的肿瘤的形成，并诱导 SNS 中观察到的基因表达模式值得注意的是，PAX3-MAML3 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，肿瘤形成。异种移植的表达 PAX3-MAML3 的细胞在雌性受体小鼠中明显大于雄性受体小鼠。原发性 SNS 肿瘤的转录组分析证实了体内雌激素反应特征的激活。这些结果使我们相信 ER 在 PAX3-MAML3 的致病活性中发挥着重要作用在 SNS 中，这构成了这种癌症的性别二态性本质。雌激素可能作为毁容手术的补充甚至替代方法这种高度侵袭性癌症的治疗将通过以下方面进行测试：目标 I：确定作用。 ER 在体外和体内 PAX3-MAML3 发病机制中的作用我们将使用以下方法检查 ER 失活的影响 CRISPR 和抗雌激素药物（即他莫昔芬、氟维司群和芳香酶抑制剂）的多个方面 PAX3-MAML3 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， (B) 在体内，我们将测试 ER 和雌激素的贡献。我们将评估 ER 失活的影响（通过 CRISPR-）。介导的 ERα/ERβ 消耗或通过用抗雌激素药物治疗小鼠），比较肿瘤生长目标 II：确定 PAX3-MAML3 和 ER 如何协调基因表达驱动侵入性生长我们将：(A) 通过 RNA-seq 定义 PAX3-MAML3 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 介导的删除 (B) 定义 PAX3-MAML3 和 ER 染色质占用景观以获得机制了解 PAX3-MAML3 和 ER 如何协调该转录程序和 ChIP- seq 数据集将被交叉以区分 PAX3-MAML3 和 ER 的直接目标基因和间接目标基因，并且深入了解转录的协同效应。这种综合分析的长期目标是促进。鉴定 PAX3-MAML3/ER 靶点及其下游通路，这对于转化的侵袭性表型，最终可能作为额外的治疗靶点。！