Quantitative Analysis of Autophagy in Single Cells: Differential Response to Amino Acid and Glucose Starvation.
Quantitative Analysis of Autophagy in Single Cells: Differential Response to Amino Acid and Glucose Starvation.
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单细胞自噬的定量分析：对氨基酸和葡萄糖饥饿的差异反应。
DOI：
10.1101/2023.12.01.569679
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
MacKeigan,JeffreyP
中科院分区：
文献类型：
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作者：
Martin,KatieR;Celano,StephanieL;Sheldon,RyanD;Jones,RussellG;MacKeigan,JeffreyP
自噬是一种高度保守的细胞内循环过程，细胞质内容物通过该过程在溶酶体中降解。这一过程发生在低水平的组成;然而，它是诱导强烈的应激反应，特别是饥饿的关键营养素，如氨基酸和葡萄糖。也就是说，这些输入的相对贡献是模糊的，许多饥饿介质定义不清或缺乏多种营养素。在这里，我们试图在正常生长条件下以及在特定氨基酸或葡萄糖饥饿的培养基中，在多个阶段和单个活细胞中生成自噬的定量目录。我们发现，在U2 OS细胞中，自噬是由氨基酸饥饿而不是葡萄糖饥饿诱导的，并且MTORC 1介导的ULK 1调节和自噬与氨基酸水平紧密相关。虽然自噬在氨基酸饥饿期间立即参与，但在持续饥饿期间自噬基因的转录上调标记的时期期间发生增强的反应。最后，我们证明了当营养恢复时，细胞立即恢复到其初始的低自噬状态，突出了自噬与环境条件之间的动态关系。除了在这里分享我们的发现外，我们还为其他对数学建模或以其他方式探索群体中单个细胞自噬感兴趣的人提供了高质量的数据资源。
Autophagy is a highly conserved, intracellular recycling process by which cytoplasmic contents are degraded in the lysosome. This process occurs at a low level constitutively; however, it is induced robustly in response to stressors, in particular, starvation of critical nutrients such as amino acids and glucose. That said, the relative contribution of these inputs is ambiguous and many starvation medias are poorly defined or devoid of multiple nutrients. Here, we sought to generate a quantitative catalog of autophagy across multiple stages and in single, living cells under normal growth conditions as well as in media starved specifically of amino acids or glucose. We found that autophagy is induced by starvation of amino acids, but not glucose, in U2OS cells, and that MTORC1-mediated ULK1 regulation and autophagy are tightly linked to amino acid levels. While autophagy is engaged immediately during amino acid starvation, a heightened response occurs during a period marked by transcriptional upregulation of autophagy genes during sustained starvation. Finally, we demonstrated that cells immediately return to their initial, low-autophagy state when nutrients are restored, highlighting the dynamic relationship between autophagy and environmental conditions. In addition to sharing our findings here, we provide our data as a high-quality resource for others interested in mathematical modeling or otherwise exploring autophagy in individual cells across a population.