Nanosensor-based monitoring of autophagy-associated lysosomal acidification in vivo
Nanosensor-based monitoring of autophagy-associated lysosomal acidification in vivo
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DOI：
10.1038/s41589-023-01364-9
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发表时间：
2023-06-15
影响因子：
14.8
通讯作者：
Heller, Daniel A.
中科院分区：
文献类型：
--
作者：
Kim, Mijin;Chen, Chen;Heller, Daniel A.
自噬是一种具有重要功能的细胞过程，其驱动神经退行性疾病和癌症。溶酶体过度酸化是自噬的标志。目前，溶酶体pH值是通过细胞培养中的荧光探针来测量的，但现有的方法不允许定量、瞬时或体内测量。在本研究中，我们开发了近红外光学纳米传感器，使用有机色心（碳纳米管上的共价sp（3）缺陷）来测量活细胞和体内自噬介导的内溶酶体过度酸化。纳米传感器定位于溶酶体，其中发射带响应于局部pH值的变化，使空间，动态和定量映射的细微变化，在溶酶体pH值。使用传感器，我们观察到细胞和肿瘤内的过度酸化管理mTORC 1和V-ATP酶调节剂，揭示溶酶体酸化反映的动力学S6 K去磷酸化和LC 3B脂化，而偏离p62降解。该传感器能够对自噬-溶酶体途径进行瞬时和体内监测。
Autophagy is a cellular process with important functions that drive neurodegenerative diseases and cancers. Lysosomal hyperacidification is a hallmark of autophagy. Lysosomal pH is currently measured by fluorescent probes in cell culture, but existing methods do not allow for quantitative, transient or in vivo measurements. In the present study, we developed near-infrared optical nanosensors using organic color centers (covalent sp(3) defects on carbon nanotubes) to measure autophagy-mediated endolysosomal hyperacidification in live cells and in vivo. The nanosensors localize to the lysosomes, where the emission band shifts in response to local pH, enabling spatial, dynamic and quantitative mapping of subtle changes in lysosomal pH. Using the sensor, we observed cellular and intratumoral hyperacidification on administration of mTORC1 and V-ATPase modulators, revealing that lysosomal acidification mirrors the dynamics of S6K dephosphorylation and LC3B lipidation while diverging from p62 degradation. This sensor enables the transient and in vivo monitoring of the autophagy-lysosomal pathway.