Lysine-selective molecular tweezers are cell penetrant and concentrate in lysosomes.
Lysine-selective molecular tweezers are cell penetrant and concentrate in lysosomes.
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DOI：
10.1038/s42003-021-02603-2
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发表时间：
2021-09-14
影响因子：
5.9
通讯作者：
Bitan G
中科院分区：
文献类型：
--
作者：
Li Z;Siddique I;Hadrović I;Kirupakaran A;Li J;Zhang Y;Klärner FG;Schrader T;Bitan G
赖氨酸选择性分子钳是一种很有前途的抗蛋白质病、病毒感染和细菌生物膜的候选药物。尽管在多种细胞和动物模型中证明了它们的功效，但关于它们的作用机制的重要问题，包括细胞迁移率和细胞内分布，迄今为止尚未得到回答。回答这些问题的主要障碍是迄今为止测试的主要化合物（称为CLR 01）的低固有荧光。在这里，我们解决这些问题，使用新的荧光标记的分子钳衍生物。我们表明，这些化合物在神经元和星形胶质细胞的内化，至少部分通过动力蛋白依赖的内吞作用。此外，我们证明了分子钳迅速集中在酸性隔间，主要是溶酶体。分子钳在溶酶体中的积累可以通过内体-溶酶体途径和通过自噬-溶酶体途径发生。此外，通过可视化分子钳，溶酶体和tau聚集体的共定位，我们表明溶酶体可能是分子钳的细胞内抗淀粉样蛋白活性的主要位点。这些发现对分子镊子在体内的作用机制具有重要意义，解释了低剂量化合物的给药如何在需要时达到高有效浓度，并支持这些化合物作为目前无法治愈的蛋白质病药物的发展。Li等设计并合成了荧光标记的分子钳，以探索它们在脑细胞（包括神经元和星形胶质细胞）中的内化、细胞内定位和内吞机制。作者发现，MT在神经元和星形胶质细胞中内化，至少部分通过动力蛋白依赖性内吞作用，并迅速集中在酸性隔室，主要是溶酶体。
Lysine-selective molecular tweezers are promising drug candidates against proteinopathies, viral infection, and bacterial biofilm. Despite demonstration of their efficacy in multiple cellular and animal models, important questions regarding their mechanism of action, including cell penetrance and intracellular distribution, have not been answered to date. The main impediment to answering these questions has been the low intrinsic fluorescence of the main compound tested to date, called CLR01. Here, we address these questions using new fluorescently labeled molecular tweezers derivatives. We show that these compounds are internalized in neurons and astrocytes, at least partially through dynamin-dependent endocytosis. In addition, we demonstrate that the molecular tweezers concentrate rapidly in acidic compartments, primarily lysosomes. Accumulation of molecular tweezers in lysosomes may occur both through the endosomal-lysosomal pathway and via the autophagy-lysosome pathway. Moreover, by visualizing colocalization of molecular tweezers, lysosomes, and tau aggregates we show that lysosomes likely are the main site for the intracellular anti-amyloid activity of molecular tweezers. These findings have important implications for the mechanism of action of molecular tweezers in vivo, explaining how administration of low doses of the compounds achieves high effective concentrations where they are needed, and supporting the development of these compounds as drugs for currently cureless proteinopathies. Li et al. design and synthesize fluorescently labeled molecular tweezers to explore their internalization, intracellular localization, and mechanism of endocytosis in brain cells including neurons and astrocytes. The authors find that MTs are internalized in neurons and astrocytes, at least partially through dynamin-dependent endocytosis, and concentrate rapidly in acidic compartments, primarily lysosomes.
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DOI：
10.1007/978-1-4939-7811-3_24
发表时间：
2018
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Malik R;Di J;Nair G;Attar A;Taylor K;Teng E;Klärner FG;Schrader T;Bitan G
通讯作者：
Bitan G
影响因子：
5
作者：
Mason AJ;Hurst I;Malik R;Siddique I;Solomonov I;Sagi I;Klärner FG;Schrader T;Bitan G
通讯作者：
Bitan G
影响因子：
14.5
作者：
Collin, Ludovic;Bohrmann, Bernd;Grueninger, Fiona
通讯作者：
Grueninger, Fiona
影响因子：
15
作者：
Fokkens, M;Schrader, T;Klärner, FG
通讯作者：
Klärner, FG
影响因子：
12.6
作者：
Hu YB;Dammer EB;Ren RJ;Wang G
通讯作者：
Wang G