Lysosome-targeting chimaeras for degradation of extracellular proteins.
Lysosome-targeting chimaeras for degradation of extracellular proteins.
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DOI：
10.1038/s41586-020-2545-9
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发表时间：
2020-08
期刊：
影响因子：
64.8
通讯作者：
Bertozzi CR
中科院分区：
文献类型：
--
作者：
Banik SM;Pedram K;Wisnovsky S;Ahn G;Riley NM;Bertozzi CR
大多数针对单个蛋白质的治疗依赖于与目标蛋白质的特定活性调节相互作用，例如，酶抑制或配体阻断。然而，几种主要的治疗相关蛋白具有未知或不可接近的活性谱，因此不能被这种策略靶向。蛋白质降解平台，如蛋白水解靶向嵌合体（PROTACs）和其他（例如，dTAGs, Trim-Away，伴侣介导的自噬靶向和SNIPERs）已经开发出来，用于通常难以靶向的蛋白质；然而，这些方法涉及到对细胞内蛋白质降解机制的操作，因此基本上仅限于含有配体可以结合并募集必要细胞成分的细胞质结构域的蛋白质。细胞外蛋白和膜相关蛋白——40%的蛋白质编码基因的产物——是癌症、衰老相关疾病和自身免疫性疾病的关键因素，因此，选择性降解这些蛋白的一般策略有可能改善人类健康。在这里，我们利用结合细胞表面溶酶体穿梭受体和靶蛋白的细胞外结构域的偶联物，建立了细胞外和膜相关蛋白的靶向降解。这些最初的溶酶体靶向嵌合体，我们称之为lytac，由一个小分子或抗体融合到化学合成的糖肽配体上，这些糖肽配体是不依赖阳离子的甘露糖-6-磷酸受体（CI-M6PR）的激动剂。我们使用lytac开发了一种CRISPR干扰筛选，揭示了细胞系中ci - m6pr介导的货物内化的生化途径，并揭示了作为该途径先前未被识别但必不可少的组成部分的胞囊复合物。我们通过治疗相关蛋白的降解证明了该平台的范围，包括载脂蛋白E4、表皮生长因子受体、CD71和程序性死亡配体1。我们的研究结果建立了一个模块化的策略来指导分泌蛋白和膜蛋白的溶酶体降解，对生化研究和治疗具有广泛的意义。
The majority of therapies that target individual proteins rely on specific activitymodulating interactions with the target protein–for example, enzyme inhibition or ligand blocking. However, several major classes of therapeutically relevant proteins have unknown or inaccessible activity profiles and so cannot be targeted by such strategies. Protein-degradation platforms such as proteolysis-targeting chimaeras (PROTACs) and others (for example, dTAGs, Trim-Away, chaperone-mediated autophagy targeting and SNIPERs) have been developed for proteins that are typically difficult to target; however, these methods involve the manipulation of intracellular protein degradation machinery and are therefore fundamentally limited to proteins that contain cytosolic domains to which ligands can bind and recruit the requisite cellular components. Extracellular and membrane-associated proteins—the products of 40% of all protein-encoding genes–are key agents in cancer, ageing-related diseases and autoimmune disorders, and so a general strategy to selectively degrade these proteins has the potential to improve human health. Here we establish the targeted degradation of extracellular and membrane-associated proteins using conjugates that bind both a cell-surface lysosome-shuttling receptor and the extracellular domain of a target protein. These initial lysosome-targeting chimaeras, which we term LYTACs, consist of a small molecule or antibody fused to chemically synthesized glycopeptide ligands that are agonists of the cation-independent mannose-6-phosphate receptor (CI-M6PR). We use LYTACs to develop a CRISPR interference screen that reveals the biochemical pathway for CI-M6PR-mediated cargo internalization in cell lines, and uncover the exocyst complex as a previously unidentified–but essential–component of this pathway. We demonstrate the scope of this platform through the degradation of therapeutically relevant proteins, including apolipoprotein E4, epidermal growth factor receptor, CD71 and programmed death-ligand 1. Our results establish a modular strategy for directing secreted and membrane proteins for lysosomal degradation, with broad implications for biochemical research and for therapeutics.
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影响因子：
25
作者：
Fan, Xuelai;Jin, Wu Yang;Lu, Jie;Wang, Jin;Wang, Yu Tian
通讯作者：
Wang, Yu Tian
影响因子：
14.8
作者：
通讯作者：
--
影响因子：
7.7
作者：
Huang, Yongjian;Bharill, Shashank;Kuriyan, John
通讯作者：
Kuriyan, John
影响因子：
4.4
作者：
Cox, Juergen;Neuhauser, Nadin;Mann, Matthias
通讯作者：
Mann, Matthias
DOI：
10.1073/pnas.1516127112
发表时间：
2015-10-13
影响因子：
11.1
作者：
Kramer, Jessica R.;Onoa, Bibiana;Bertozzi, Carolyn R.
通讯作者：
Bertozzi, Carolyn R.