The Role of the Fructose-1,6-Bisphoshatase 2 and c-Myc Interaction in Sarcoma Progression

果糖-1,6-双磷酸酶 2 和 c-Myc 相互作用在肉瘤进展中的作用

• 批准号: 10536204
• 负责人: Bailey Nance
• 金额: $ 4.68万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10536204
• 关键词: Adult; Affect; Alanine; Alleles; Attenuated; Binding; Binding Sites; Biogenesis; Biological Assay; CDK4 gene; Cartilage; Cell Cycle; Cell Cycle Regulation; Cell Fractionation; Cell Nucleus; Cells; Co-Immunoprecipitations; Complement; Complex; Connective Tissue; Cytoplasm; Data; Dependence; Disease; Down-Regulation; Enzymes; Excision; Exhibits; Exposure to; Fatty acid glycerol esters; Fructose; Fructose-1,6-Bisphosphatase; Gene Expression; Genetic; Genetic Transcription; Gluconeogenesis; Glucose; Growth; Heterogeneity; Histologic; Hypoxia; In Vitro; Malignant Neoplasms; Maps; Measures; Mediating; Mesenchymal Cell Neoplasm; Metabolic; Metabolism; Methods; Mitochondria; Modeling; Molecular; Molecular Conformation; Mus; Muscle; N-terminal; Nuclear; Nuclear Export; Nuclear Translocation; Operative Surgical Procedures; Oxidative Phosphorylation; Patients; Play; Protein Isoforms; Proteins; Recurrence; Research; Resistance; Role; Scanning; Signal Transduction; Site; Site-Directed Mutagenesis; Soft tissue sarcoma; Stimulus; Structure; Therapeutic; analytical ultracentrifugation; aurora kinase A; c-myc Genes; cdc Genes; chemotherapy; combat; confocal imaging; conventional therapy; dimer; effective therapy; epigenetic silencing; experimental study; functional outcomes; genetic signature; glucose metabolism; glucose uptake; improved; in vivo; inhibitor; insight; mouse model; mutant; novel; novel therapeutics; prediction algorithm; promoter; protein protein interaction; restoration; sarcoma; sedimentation velocity; small molecule inhibitor; transcriptome sequencing; treatment response; tumor; tumor growth

PROJECT SUMMARY Soft tissue sarcomas (STSs) are diverse mesenchymal tumors that occur within connective tissues such as muscle, fat, and cartilage. STS is a highly heterogeneous malignancy with over 70 genetic and histological subtypes. In addition, high rates of recurrence and lack of effective treatment for patients emphasize the need to identify novel therapeutic vulnerabilities common to multiple STS subtypes. Metabolically, STS tumors consistently exhibit high rates of glucose uptake and robust hypoxia gene signatures in patients. Therefore, we focused on fructose-1,6-bisphosphatase (FBP), a rate-limiting enzyme in gluconeogenesis, to combat the typical glycolytic dependence of STS. In previous studies, we showed that FBP2, the muscle-specific isoform of FBP, is severely downregulated in several prevalent STS subtypes. Surprisingly, FBP2 re-expression in STS in vivo opposes sarcoma progression through two spatially distinct mechanisms in the cytoplasm and the nucleus. FBP2 restoration decreases cytosolic glycolytic flux while nuclear FBP2 directly binds to c-Myc, a transcriptional regulator of growth and metabolism. This interaction attenuates c-Myc-dependent expression of TFAM, a key regulator of mitochondrial biogenesis, thereby inhibiting oxidative phosphorylation. These mechanisms, at least in part, contribute to the ability of re-expressed FBP2 to suppress tumor growth in murine STS models. However, my preliminary data suggest that nuclear FBP2 may also directly regulate other c-Myc cell cycle target genes CDK4 and AURKA to contribute to STS suppression. Moreover, how FBP2 regulates c-Myc and the sites of FBP2/c-Myc binding are not known. Recent studies suggest that FBP2 oligomerization and conformation may also play an important role in FBP2's nuclear-to-cytosolic shuttling, and reveal new binding sites for other proteins in the nucleus like c-Myc. Therefore, I hypothesize that the exposed N-terminal regions of tetrameric FBP2 complexes bind directly to c-Myc, contributing to STS suppression by inhibiting both TFAM and cell cycle regulators. In Aim 1, I will map the sites of interaction between FBP2 and c-Myc and determine whether the tetrameric state of FBP2 is necessary for binding. In Aim 2, I will explore the functional outcomes of the FBP2/c-Myc interaction in sarcoma, including the regulation of cell cycle genes AURKA and CDK4. Together, this proposal will provide more insight into the recently discovered nuclear functions of FBP2 to create novel therapies for a diverse set of genetically heterogeneous sarcomas.

项目摘要 软组织肉瘤（STS）是发生在结缔组织中的多种间叶肿瘤， 肌肉脂肪和软骨STS是一种高度异质性的恶性肿瘤， 亚型此外，高复发率和患者缺乏有效治疗强调了需要 以确定多种STS亚型共有的新的治疗弱点。代谢方面，STS肿瘤 患者始终表现出高葡萄糖吸收率和强大的缺氧基因特征。所以我们 专注于果糖-1，6-二磷酸酶（FBP），这是一种在植物异生中的限速酶，以对抗典型的 STS的糖酵解依赖性。在以前的研究中，我们发现，肌肉特异性的FBP亚型FBP2， 在几种流行的STS亚型中严重下调。令人惊讶的是，体内STS中的FBP2再表达 通过细胞质和细胞核中两种空间上不同的机制阻止肉瘤进展。FBP2 恢复减少胞质糖酵解通量，而核FBP2直接结合到c-Myc，一个转录因子。 生长和新陈代谢的调节剂。这种相互作用减弱了TFAM的c-Myc依赖性表达，TFAM是一个关键的 线粒体生物发生的调节剂，从而抑制氧化磷酸化。这些机制，至少 在某种程度上，有助于重新表达的FBP2在鼠STS模型中抑制肿瘤生长的能力。然而，在这方面， 我的初步数据表明，核FBP2也可能直接调控其他c-Myc细胞周期靶基因 CDK4和AURKA有助于STS抑制。此外，FBP2如何调节c-Myc和 FBP2/c-Myc结合未知。最近的研究表明，FBP2的寡聚化和构象可能 在FBP2的核-胞质穿梭中也起重要作用，并揭示了其他蛋白质的新结合位点 像c-Myc一样。因此，我假设四聚体FBP2的暴露的N-末端区域 复合物直接与c-Myc结合，通过抑制TFAM和细胞增殖来抑制STS。 循环调节器。在目标1中，我将绘制FBP2和c-Myc之间相互作用的位点，并确定是否 FBP2的四聚体状态是结合所必需的。在目标2中，我将探讨 FBP2/c-Myc在肉瘤中的相互作用，包括对细胞周期基因AURKA和CDK4的调节。在一起， 该提案将为最近发现的FBP2核功能提供更多见解，以创造新的 一系列遗传异质性肉瘤的治疗方法。