Spatio-temporal regulation of mTORC1 signaling in normal and disease states

正常和疾病状态下 mTORC1 信号传导的时空调节

基本信息

批准号：
10174962
负责人：
Roberto Zoncu
金额：
$ 31.4万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
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：
2019-07-25 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10174962
关键词：
Affect Affinity Amino Acids Attenuated Autophagocytosis Binding Biochemical Cancer Cell Growth Catabolic Process Cells Cellular Metabolic Process Chemicals Complex Coupled Covalent Interaction Cytoplasm Development Diabetes Mellitus Disease Dissociation Engineering FRAP1 gene Generations Glucose Goals Growth Growth Factor Guanosine Image In Vitro Knowledge Lead Lipids Lymphoma Lysosomes Malignant Neoplasms Maps Measures Membrane Metabolic Metabolism Molecular Mutagenesis Mutation Nerve Degeneration Non-Insulin-Dependent Diabetes Mellitus Nucleotide Biosynthesis Nucleotides Nutrient Oncogenic Output Oxygen Process Protein Biosynthesis Protein Kinase Regulation Renal Cell Carcinoma Research Role Signal Transduction Site Stimulus Structure Surface System Testing Therapeutic Work base comparative efficacy deletion analysis detection of nutrient driving force falls human disease in vitro Assay inhibitor/antagonist innovation insight kinase inhibitor lipid biosynthesis mutant novel novel strategies programs reconstitution recruit repaired response scaffold screening small molecule spatiotemporal tool

项目摘要

PROJECT SUMMARY The molecular mechanisms through which cells sense nutrients remain largely unknown, but their elucidation is key to our understanding of metabolic regulation both in normal and disease states. At the center of nutrient sensing and growth regulation is an ancient protein kinase known as the mechanistic Target of Rapamycin Complex 1 (mTORC1). In response to the combined action of metabolic inputs such as nutrients, growth factors, energy and oxygen, mTORC1 translocates from the cytoplasm to the surface of lysosomes, where it becomes activated. Accumulating evidence indicates that aberrant mTORC1 activation at the lysosome could be a driving force in diseases ranging from cancer to type-2 diabetes to neurodegeneration. Lysosomal translocation and activation of mTORC1 requires the heterodimeric Rag guanosine triphosphatases (GTPases), which together with the pentameric Ragulator complex, form a nutrient-regulated scaffolding complex that physically anchors mTORC1 to the lysosomal surface. Combining dynamic imaging in cells with biochemical reconstitution and structural approaches, we recently discovered that the Ragulator-Rag complex is not static but is rather actively remodeled by nutrients, leading to spatial cycling of the Rag GTPases between the lysosomal surface and the cytoplasm. In turn, Rag cycling places a limit on the efficiency of mTORC1 capture and may facilitate its inactivation when nutrient levels fall. Importantly, Rag cycling is altered by cancer-specific mutations that affect mTORC1 signaling. Based on these findings, we hypothesize that spatial-temporal regulation of mTORC1 scaffolding is a novel and unrecognized mechanism to modulate the potency and selectivity of mTORC1 signaling responses, and that its disruption may drive the aberrant growth of mTORC1-driven cancers, including renal cell carcinoma and lymphoma. We will test this hypothesis via two highly complementary and innovative research aims. First, we will employ structure-guided mutagenesis to dissect the mechanisms that govern the assembly of the mTORC1- scaffolding complex in response to changing nutrient inputs. Second, we will characterize the mechanism of action of new-generation compounds we recently discovered, which block the assembly of the lysosomal mTORC1 scaffolding complex, and determine their ability to inhibit the metabolism and growth of mTORC1- driven cancers. Collectively, the proposed studies will generate new knowledge on the spatial-temporal regulation of mTORC1 signaling, and point the way to novel strategies to manipulate mTORC1 signaling in both normal and disease states.

项目摘要细胞感知养分的分子机制在很大程度上未知，但它们的分子机制阐明是我们对正常和疾病状态中代谢调节的理解的关键。在中心营养感应和生长调节是一种古老的蛋白激酶，称为雷帕霉素复合物1（MTORC1）。响应代谢输入（例如营养素）的联合作用，生长因子，能量和氧，mtorc1从细胞质转移到溶酶体表面，它被激活的地方。积累的证据表明，MTORC1在溶酶体可能是从癌症到2型糖尿病再到神经变性的疾病的驱动力。 MTORC1的溶酶体易位和激活需要异二聚体RAG鸟苷三磷酸酶（GTPases）与五聚式间发仪配合物一起形成营养素调节将MTORC1物理锚定在溶酶体表面的脚手架复合物。结合动态成像具有生化重构和结构方法的细胞，我们最近发现ragulator-rag 复合物不是静态的，而是由营养物质积极重塑，导致抹布的空间循环溶酶体表面和细胞质之间的GTP酶。反过来，抹布骑自行车对 MTORC1捕获的效率，并在养分水平下降时可能促进其失活。重要的是，抹布循环会因影响MTORC1信号传导的癌症特异性突变而改变。基于这些发现，我们假设MTORC1脚手架的时空调节是一种新颖且未认可的机制调节MTORC1信号响应的效力和选择性，并且其破坏可能会驱动 MTORC1驱动的癌症的异常生长，包括肾细胞癌和淋巴瘤。我们将通过两个高度互补和创新的研究目的来检验这一假设。首先，我们会的采用结构引导的诱变来剖析控制MTORC1-组装的机制脚手架复合物响应不断变化的养分输入。其次，我们将表征我们最近发现的新一代化合物的作用，该化合物阻断了溶酶体的组装 MTORC1脚手架复合物，并确定它们抑制MTORC1-的代谢和生长的能力驱动的癌症。总的来说，拟议的研究将产生有关时空调节的新知识 MTORC1信号传导，并指向采取新型策略来操纵MTORC1信号的方法疾病状态。