K-Ras sumoylation in cell proliferation and transformation

细胞增殖和转化中的 K-Ras 修饰

• 批准号: 9924462
• 负责人: Yuan Chen
• 金额: $ 49.75万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-15 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9924462
• 关键词: Acetylation; Animal Model; Antineoplastic Agents; Autophagocytosis; Biochemical; Biological Models; Cell Proliferation; Cell Survival; Cell physiology; Cells; Development; Ectopic Expression; Enzymes; Excision; Family; Glycine; Growth; Guanosine Triphosphate Phosphohydrolases; In Vitro; KRAS2 gene; Lead; Lysine; MAP3K1 gene; MEKs; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of pancreas; Mammalian Cell; Mediating; Methylation; Modification; Molecular; Monomeric GTP-Binding Proteins; Monoubiquitination; Mutation; Oncogenic; Pancreas; Patients; Phosphorylation; Post-Translational Protein Processing; Process; Proteins; RAS genes; Regulatory Pathway; Research; Resistance; Role; Series; Signal Transduction; Sumoylation Pathway; System; Testing; Tumor-Derived; Ubiquitin; Ubiquitination; Xenograft procedure; anti-cancer; base; cancer therapy; cell growth; cell immortalization; cell motility; cell transformation; design; farnesylation; gene product; in vivo; inhibitor/antagonist; insight; interest; isopeptidase; migration; mouse model; mutant; novel; palmitoylation; pancreatic cancer cells; pre-clinical; protein protein interaction; ral Guanine Nucleotide Exchange Factor; ras GTPase-Activating Proteins; ras Proteins; small molecule inhibitor; sulfoenolpyruvate; targeted treatment; therapeutic development; therapy development; tumor xenograft; tumorigenesis; ubiquitin-protein ligase

ABSTRACT Ras proteins are GTPases that activate multiple signaling cascades, which modulate cell survival, proliferation, migration, and differentiation. Ras proteins are regulated by almost all known post-translational modifications, including farnesylation, palmitoylation, phosphorylation, ubiquitination, and acetylation. However, it remains unknown whether Ras proteins are modified by sumoylation. We recently discovered that all three forms of Ras protein are SUMO3-modified. Our further studies reveal that lysine 42 (K42) is the primary residue for sumoylation, whereas PIASγ and MEKK1 are candidate E3 ligases that sumoylate Ras. Sumoylation is essential for activating K-Ras and its downstream signaling. Furthermore, we have discovered a series of SUMO E1 inhibitors that target SUMO conjugation both in vitro and in vivo. The lead compounds 27A and SB-264 potently inhibit sumoylation, but not ubiquitination or neddylation. Significantly, these compounds display excellent bioactivities toward killing transformed pancreatic cells with K-Ras mutations at glycine 12 or glycine 13 and suppress their migration. Given the role of sumoylation in controlling functions of key cellular regulators and the paramount importance of K- Ras in cell proliferation and transformation, we hypothesize that sumoylation promotes K-Ras activity and that the SUMO regulatory pathway is an excellent target for development of anti- cancer drugs. Three Specific Aims are proposed to test the validity of the hypothesis. We will determine the mechanism by which sumoylation regulates K-Ras activation, elucidate the regulatory network that controls K-Ras sumoylation, and investigate the efficacy of SUMO E1 inhibitors in suppressing proliferation of pancreatic cancer growth in preclinical mouse models. Further characterization of K-Ras sumoylation promises to have a profound impact on our understanding of the fundamental cellular signaling mechanisms that regulate cell growth. Given the discovery of novel SUMO E1 inhibitors, our proposed studies will help to accelerate the development of therapeutic applications that target ubiquitin-like modifications. !

抽象的 Ras 蛋白是 GTP 酶，可激活多个信号级联，从而调节细胞 生存、增殖、迁移和分化。 Ras 蛋白受几乎所有 已知的翻译后修饰，包括法呢基化、棕榈酰化、磷酸化、 泛素化和乙酰化。然而，Ras蛋白是否被修饰仍不清楚 通过苏木化作用。我们最近发现所有三种形式的 Ras 蛋白均经过 SUMO3 修饰。 我们的进一步研究表明，赖氨酸 42 (K42) 是苏酰化的主要残基，而 PIASγ 和 MEKK1 是可苏酰化 Ras 的候选 E3 连接酶。苏酰化作用对于 激活 K-Ras 及其下游信号传导。此外，我们还发现了一系列 SUMO E1 抑制剂在体外和体内均以 SUMO 结合为目标。铅化合物 27A和SB-264有效抑制苏酰化，但不抑制泛素化或neddylation。显著地， 这些化合物显示出优异的生物活性，可杀死转化的胰腺细胞 K-Ras 在甘氨酸 12 或甘氨酸 13 处发生突变并抑制其迁移。鉴于角色 sumoylation 在控制关键细胞调节因子的功能中以及 K-的至关重要性 Ras 在细胞增殖和转化中的作用，我们假设 sumoylation 促进 K-Ras 活性并且 SUMO 调控途径是开发抗- 癌症药物。提出了三个具体目标来检验假设的有效性。我们将 确定 sumoylation 调节 K-Ras 激活的机制，阐明 控制 K-Ras sumoylation 的调控网络，并研究 SUMO E1 的功效 抑制临床前小鼠模型中胰腺癌生长增殖的抑制剂。 K-Ras 苏酰化的进一步表征有望对我们的研究产生深远的影响 了解调节细胞生长的基本细胞信号传导机制。给定 新型 SUMO E1 抑制剂的发现，我们提出的研究将有助于加速 开发针对泛素样修饰的治疗应用。 ！