The role and regulation of Hippo pathway in sarcomagenesis

Hippo通路在肉瘤发生中的作用及调控

• 批准号: 10738329
• 负责人: Tzipora Sarah Karin Eisinger
• 金额: $ 7.88万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 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/10738329
• 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; KRASG12D; Knowledge; Libraries; Lysine; Malignant Fibrous Histiocytoma; Malignant Neoplasms; Mediating; Mediator; 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; Proliferating; 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; 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

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.

项目总结/摘要 软组织肉瘤是一种侵袭性的间叶恶性肿瘤， 全球每年。与上皮癌不同，新的靶向治疗在上皮癌中具有显著的 对患者生存率的影响，包括肉瘤在内的间叶肿瘤的治疗方法还没有 25年来发生了巨大变化。我们最近的研究表明，Hippo通路的失调增强了 侵袭性肌肉肿瘤中的肉瘤发生，未分化多形性肉瘤（UPS）。UPS是一个 常见于成人肌肉组织的诊断和转移性肉瘤亚型。我们有 观察到血管动素（AMOT）的损失，一个重要的介导海马相关的生长限制，是必要的 UPS肉瘤形成AMOT在分化的人肌肉组织中高度表达，但在分化的人肌肉组织中沉默。 UPS和其他肉瘤。AMOT的p130亚型的异位再表达显著抑制肉瘤 体外细胞增殖。这一发现与AMOT在癌细胞中唯一已知的功能一致， 是隔离Hippo途径效应子YAP 1并促进其降解。YAP 1是一种促增殖因子， 在UPS的本地小鼠模型中缺失的转录调节因子显著降低 肿瘤发生总之，这些数据表明，AMOT的缺失促进了YAP介导的肉瘤发生。 肌肉来源的UPS。接下来，我们通过微阵列研究了UPS中YAP 1表达的下游效应。 对照和Yap 1缺陷型鼠肿瘤的基因表达研究。我们发现Yap 1调控NF-κB 通过抑制Usp 31的表达，Usp 31是NF-κB活性的负调节剂，从而在UPS中发挥信号传导作用。此外，委员会认为， 使用患者样品的ChIP-seq，我们发现NF-κB信号在人UPS中显著上调。 与这些发现一致，UPS细胞增殖对NF-κB抑制高度敏感。基于这些 具体目标1中的调查结果，我们将确定如何在UPS中控制AMOT损耗，以及是否需要此过程 用于软组织肉瘤的肿瘤起始。接下来，我们将定义YAP 1抑制 Specific Aim 2中的USP 31表达。我们将研究YAP 1是否直接与启动子区结合， USP 31，阻止其转录。YAP 1的缺失恢复了USP 31的表达，USP 31是一种肽酶， 从p65上游的TRAF分子中的赖氨酸63活化特异性泛素修饰，从而 使NF-κB失活。NF-κB在正常骨骼肌祖细胞中的作用， 是促进增殖和防止分化。在具体目标3中，我们将确定哪一个YAP 1- 依赖性NF-κB靶点对于调节这些过程中的一个或两个是必需的。这个目标 一个提议是检验Hippo信号通过抑制促进肉瘤发生的假说 AMOT的表达，导致YAP 1/NF-κ B相关的增殖和分化抑制。最终 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。