Structure and Function of the SHOC2 Holophosphatase Complex in RAS-driven Cancer

SHOC2 全磷酸酶复合物在 RAS 驱动的癌症中的结构和功能

• 批准号: 10662750
• 负责人: Jason Kwon
• 金额: $ 14.72万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2023-12-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10662750
• 关键词: Acceleration; Amino Acid Substitution; Binding; Biological Assay; Biology; Biophysics; CRISPR/Cas technology; Cancer cell line; Cell Therapy; Cell membrane; Clinical; Complex; Cycloheximide; Data; Development; Drug resistance; Educational process of instructing; Enzymes; Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor; Future; Genetic Screening; Goals; Half-Life; Homologous Gene; Image; Immunofluorescence Immunologic; Immunoprecipitation; In Vitro; KRAS2 gene; Knowledge; Laboratories; Lead; Libraries; Lysine; MEKs; Malignant Neoplasms; Malignant neoplasm of pancreas; Maps; Mass Spectrum Analysis; Measures; Mediating; Mitogen-Activated Protein Kinase Inhibitor; Mitogen-Activated Protein Kinases; Modification; Muscle; Mutate; Mutation; Oncogenic; Organoids; Outcome Study; Pathway interactions; Patients; Phase; Positioning Attribute; Post-Translational Protein Processing; Protein Dephosphorylation; Protein Family; Protein Inhibition; Proteins; Proteomics; RAF1 gene; RAS driven cancer; Recurrence; Reporter; Reporting; Research; Resistance; Scaffolding Protein; Signal Transduction; Site; Specificity; Structural Models; Structure; Substrate Specificity; Surface; Therapeutic; Training; Ubiquitin; Ubiquitination; Variant; Work; cancer cell; crosslink; experience; inhibitor; insight; leukemia; loss of function; melanoma; member; mutant; mutation screening; novel; novel strategies; programs; protein function; protein protein interaction; protein structure function; ras Proteins; rational design; refractory cancer; resistance mechanism; response; scaffold; small molecule; structural biology; targeted treatment; therapeutic development; therapeutic target; tumor progression

Project Summary/Abstract: RAS proteins are found to be frequently mutated in clinical cases of cancer. Effective targeted therapies against RAS and its downstream pathways have been highly sought after. Among the RAS effectors, the RAF/mitogen-activated protein kinase (MAPK) cascade has been shown to be important for propagating oncogenic signaling downstream of RAS. Many promising targeted therapies have been developed that directly inhibit mutant KRAS and MAPK members but have been limited due to resistance mechanisms. SHOC2 has been recently found to be a potent synthetic lethal vulnerability in various RAS mutant contexts, including RAS-driven leukemia/melanoma and facilitates cancer resistance to numerous RAS/MAPK inhibitors. SHOC2 is a scaffold that binds RAS and PP1C to form a ternary holophosphatase complex which dephosphorylates inhibitory ‘S259’ on RAF proteins and enables MAPK signaling. To actualize SHOC2 as a therapeutic target and inform future efforts to rationally design inhibitors against the complex, a deep understanding of SHOC2 structural features that govern complex assembly and function is needed. We have now resolved the SHOC2 complex structure and have profiled the functional consequence of nearly all possible amino acid substitutions at every position of SHOC2 through deep mutational scanning. These findings highlight novel structural features of SHOC2 that may be important to therapeutically target in RAS mutant cancers. The studies outlined in this proposal will build on this work and aim to (1) define and validate the structural features of SHOC2 holophosphatase complex specificity toward RAF substrate, and (2) identify novel post-translational modifications that regulate SHOC2 function. The training I receive in the K99 phase will provide me the necessary expertise in structural biology, proteomics, and ubiquitin biology as well as equip me with teaching/lab management experience so that I can effectively lead my own independent research program. The K99/R00 will be instrumental in helping me achieve my long-term goal, to establish my laboratory focused on studying novel RAS-associated vulnerabilities and drug resistance mechanisms via systematic approaches in cell signaling and protein structure-function.

项目摘要/摘要： 发现在癌症的临床病例中，发现RAS蛋白经常突变。有效的靶向疗法 反对RAS及其下游途径之后闻起来很高。在RAS效应中， RAF/有丝分裂原激活的蛋白激酶（MAPK）级联反应已被证明对于传播很重要 RAS下游的致癌信号传导。已经开发了许多承诺的有针对性疗法 直接抑制突变的KRAS和MAPK成员，但由于电阻机制而受到限制。 最近发现SHOC2是各种RAS突变体中潜在的合成致命脆弱性 环境，包括RAS驱动的白血病/黑色素瘤，并促进癌症对众多RAS/MAPK的抗性 抑制剂。 SHOC2是一种结合Ras和Pp1c的支架，形成了三元群磷酸酶络合物，该复合酶配合物 在RAF蛋白上脱磷酸化抑制性“ S259”，并启用MAPK信号传导。将Shoc2实现为 治疗目标并为未来的努力提供理性设计抑制剂，以抵抗该综合体，这是一个深刻的 需要了解SHOC2结构特征，这些结构特征需要控制复杂的组装和功能。我们有 现在解决了Shoc2复合结构，并介绍了几乎所有的功能结果 通过深突变扫描，可能在Shoc2的每个位置取代了氨基酸的取代。 这些发现突出了Shoc2的新结构特征，这对于治疗可能很重要 在Ras突变癌中。该提案中概述的研究将以这项工作为基础，并旨在（1）定义和 验证Shoc2求合酶对RAF底物的复杂特异性的结构特征，（2） 确定新型的翻译后修饰，以调节SHOC2功能。我在K99中接受的培训 阶段还将为我提供结构生物学，蛋白质组学和泛素生物学的必要专业知识 作为我的教学/实验室管理经验，我可以有效地领导自己的独立 研究计划。 K99/R00将有助于我实现长期目标，以确定我的 实验室致力于研究新型RAS相关的脆弱性和耐药性机制 细胞信号传导和蛋白质结构功能的系统方法。