The role and regulation of Hippo pathway in sarcomagenesis

Hippo通路在肉瘤发生中的作用及调控

基本信息

批准号：
10524097
负责人：
Tzipora Sarah Karin Eisinger
金额：
$ 8.05万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-03-01 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10524097
关键词：
AMOT gene Acetylation Adipose tissue Adult Amputation Binding Binding Sites CRISPR/Cas technology Carcinoma Cell Line Cell Nucleus Cell Proliferation Cells ChIP-seq Complex Consensus DNA Binding Data Deacetylase Development Diagnosis Distal Epigenetic Process Gene Chips Gene Expression Genetic Genetic Transcription Genetically Engineered Mouse Goals Growth Histology Histones Human In Vitro Institutes KRASG12D Knowledge Libraries Lysine Malignant Fibrous Histiocytoma Malignant Neoplasms Mediating Mediator of activation protein Mesenchymal Mesenchymal Cell Neoplasm Methylation Modeling Modification Molecular Mus Muscle Muscle satellite cell Mutate Myomatous neoplasm NF1 gene Nucleic Acid Regulatory Sequences Operative Surgical Procedures Outcome Pathway interactions Patients Peptide Hydrolases Persons Process Promoter Regions Protein Isoforms Proteins Radiation Regulation Role Sampling Signal Transduction Skeletal Muscle Skeletal Muscle Neoplasm Soft tissue sarcoma Testing Therapeutic Therapeutic Intervention Tissues Transferase Ubiquitin Work base cancer cell clinically actionable inhibitor interest mouse model new therapeutic target novel p65 precursor cell predictive marker prevent progenitor programs promoter response sarcoma targeted treatment therapeutic target transcription factor tumor tumor initiation tumorigenesis

项目摘要

Project Summary/Abstract Soft tissue sarcomas are an aggressive group of mesenchymal malignancies diagnosed in 200,000 people per year worldwide. Unlike in epithelial cancers, where novel targeted therapies have had a dramatic effect on patient survival, the treatment approach for mesenchymal tumors including sarcomas has not changed significantly in 25 years. Our recent work revealed that deregulation of the Hippo pathway enhances sarcomagenesis in the aggressive muscle tumor, Undifferentiated pleomorphic sarcoma (UPS). UPS is a commonly diagnosed and metastatic sarcoma subtype frequently found in adult muscle tissues. We have observed that loss of Angiomotin (AMOT), a crucial mediator of Hippo-associated growth restriction, is required for UPS sarcomagenesis. AMOT is highly expressed in differentiated human muscle tissue but is silenced in UPS and other sarcomas. Ectopic re-expression of the p130 isoform of AMOT significantly inhibits sarcoma cell proliferation in vitro. This finding is consistent with the only known function of AMOT in cancer cells, which is to sequester the Hippo pathway effector YAP1 and facilitate its degradation. YAP1 is a pro-proliferation transcriptional regulator whose deletion in an autochthonous mouse model of UPS significantly decreased tumorigenesis. Together these data suggest that AMOT loss promotes YAP-mediated sarcomagenesis in muscle-derived UPS. We next investigated the downstream effects of YAP1 expression in UPS by microarray gene expression studies of control and Yap1-deficient murine tumors. We found that Yap1 controls NF-κB signaling in UPS by suppressing expression of Usp31, a negative regulator of NF-κB activity. Furthermore, using ChIP-seq of patient samples we found that NF-κB signaling is substantially upregulated in human UPS. Consistent with these findings, UPS cell proliferation is highly sensitive to NF-κB inhibition. Based on these findings in Specific Aim1 we will determine how AMOT loss is controlled in UPS and if this process is required for tumor initiation in soft tissue sarcomas. Next we will define the mechanism by which YAP1 suppresses USP31 expression in Specific Aim 2. We will investigate whether YAP1 directly binds to the promoter region of USP31, preventing its transcription. Loss of YAP1 restores expression of USP31, a peptidase that removes activation specific ubiquitin modifications from lysine 63 in TRAF molecules upstream of p65, thereby inactivating NF-κB. The role of NF-κB in normal skeletal muscle progenitors, the putative cell of origin of UPS, is to promote proliferation and prevent differentiation. In Specific Aim3 we will determine which YAP1- dependent NF-κB targets are necessary for regulating either or both of these processes. The goal of this proposal is to test the hypothesis that deregulated Hippo signaling promotes sarcomagenesis via suppression of AMOT, resulting in YAP1/NF-κB-associated proliferation and inhibition of differentiation. Ultimately, the purpose of these studies is to identify clinically actionable therapeutic targets to advance treatment for skeletal muscle UPS patients.

项目概要/摘要软组织肉瘤是一组侵袭性间叶性恶性肿瘤，诊断人数达 200,000 人与全球每年的上皮癌不同，新型靶向疗法取得了巨大的成功。对患者生存的影响，包括肉瘤在内的间质肿瘤的治疗方法尚未我们最近的工作表明，Hippo 通路的放松管制增强了这一点。侵袭性肌肉肿瘤中的肉瘤发生，未分化多形性肉瘤（UPS）是一种。常见于成人肌肉组织中常见的诊断和转移性肉瘤亚型。观察到血管动蛋白 (AMOT) 的丧失是必要的，血管动蛋白是 Hippo 相关生长受限的关键介质 AMOT 在分化的人类肌肉组织中高表达，但在 UPS 肉瘤发生中沉默。 AMOT 的 p130 亚型异位重新表达可显着抑制肉瘤。这一发现与 AMOT 在癌细胞中的唯一已知功能一致。是隔离 Hippo 途径效应子 YAP1 并促进其降解 YAP1 是一种促增殖剂。转录调节因子，其在 UPS 小鼠模型中的缺失显着减少这些数据共同表明，AMOT 缺失促进 YAP 介导的肉瘤发生。接下来我们通过微阵列研究了 YAP1 表达在 UPS 中的下游效应。对照和 Yap1 缺陷小鼠肿瘤的基因表达研究我们发现 Yap1 控制 NF-κB。通过抑制 Usp31（NF-κB 活性的负调节因子）的表达来调节 UPS 中的信号传导。此外，使用患者样本的 ChIP-seq，我们发现 NF-κB 信号在人类 UPS 中显着上调。与这些发现一致，UPS 细胞增殖对 NF-κB 抑制高度敏感。具体目标 1 中的调查结果我们将确定 UPS 中如何控制 AMOT 损失以及是否需要此过程接下来我们将定义 YAP1 抑制肿瘤的机制。具体目标2中USP31的表达。我们将研究YAP1是否直接结合到启动子区域阻止 USP31，YAP1 转录的丢失会恢复 USP31 的表达，USP31 是一种去除肽酶。激活 p65 上游 TRAF 分子中赖氨酸 63 的特异性泛素修饰，从而 NF-κB 失活 NF-κB 在正常骨骼肌祖细胞（UPS 的假定起源细胞）中的作用，是促进增殖并防止分化，在Specific Aim3中我们将确定哪个YAP1-。依赖的 NF-κB 靶标对于调节这两个过程中的一个或两个过程是必要的。该提案旨在检验以下假设：解除对 Hippo 信号传导的管制，通过抑制促进肉瘤发生 AMOT，导致 YAP1/NF-κB 相关增殖和分化抑制。这些研究的目的是确定临床上可行的治疗靶点，以推进骨骼肌疾病的治疗肌肉UPS患者。