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：
10408711
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/10408711
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 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.

项目概要细胞感知营养物质的分子机制仍然很大程度上未知，但它们的阐明是我们理解正常和疾病状态下代谢调节的关键。在中心营养感应和生长调节是一种古老的蛋白激酶，被称为机械靶标雷帕霉素复合物 1 (mTORC1)。为了响应营养物质等代谢输入的综合作用，生长因子、能量和氧气，mTORC1从细胞质易位到溶酶体表面，它被激活的地方。越来越多的证据表明 mTORC1 的异常激活溶酶体可能是癌症、2 型糖尿病和神经退行性疾病等疾病的驱动力。 mTORC1 的溶酶体易位和激活需要异二聚体 Rag 鸟苷三磷酸酶 (GTPase) 与五聚 Ragulator 复合物一起形成营养调节因子将 mTORC1 物理锚定到溶酶体表面的支架复合物。结合动态成像细胞与生化重建和结构方法，我们最近发现 Ragulator-Rag 复合体不是静态的，而是通过营养物质积极重塑，导致 Rag 的空间循环溶酶体表面和细胞质之间的 GTP 酶。反过来，抹布骑行也限制了 mTORC1 捕获的效率，并可能在营养水平下降时促进其失活。重要的是，拉格影响 mTORC1 信号传导的癌症特异性突变会改变循环。基于这些发现，我们假设 mTORC1 支架的时空调节是一种新颖且未被认识的机制调节 mTORC1 信号反应的效力和选择性，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标来检验这一假设。首先，我们将采用结构引导诱变来剖析控制 mTORC1- 组装的机制脚手架复合体响应不断变化的营养输入。其次，我们将描述其机制我们最近发现的新一代化合物的作用，可以阻止溶酶体的组装 mTORC1支架复合物，并测定其抑制mTORC1-代谢和生长的能力驱动癌症。总的来说，所提出的研究将产生关于时空调控的新知识 mTORC1 信号传导，并为在正常和正常情况下操纵 mTORC1 信号传导的新策略指明了道路疾病状态。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The Lysosome at the Intersection of Cellular Growth and Destruction.

溶酶体位于细胞生长和破坏的交叉点。

DOI：
10.1016/j.devcel.2020.06.010
发表时间：
2020-07-20
期刊：
Developmental cell
影响因子：
11.8
作者：
Shin HR;Zoncu R
通讯作者：
Zoncu R

NPC1-mTORC1 Signaling Couples Cholesterol Sensing to Organelle Homeostasis and Is a Targetable Pathway in Niemann-Pick Type C.

NPC1-mTORC1 信号传导将胆固醇感应与细胞器稳态结合起来，是 Niemann-Pick C 型中的一个靶向途径。

DOI：
发表时间：
2021
期刊：
影响因子：
11.8
作者：
Davis, Oliver B;Shin, Hijai R;Lim, Chun;Wu, Emma Y;Kukurugya, Matthew;Maher, Claire F;Perera, Rushika M;Ordonez, M Paulina;Zoncu, Roberto
通讯作者：
Zoncu, Roberto

Picking the arginine lock on PQLC2 cycling.

撬开 PQLC2 循环中的精氨酸锁。