High-Throughput Screens To Identify Autophagy Inducers That Function by Disrupting Beclin 1/Bcl-2 Binding,ACS Chemical Biology

当前位置： X-MOL 学术 › ACS Chem. Biol. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

High-Throughput Screens To Identify Autophagy Inducers That Function by Disrupting Beclin 1/Bcl-2 Binding
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2018-06-07 00:00:00 , DOI: 10.1021/acschembio.8b00421
Wei-Chung Chiang ₁ , Yongjie Wei _{1,

2} , Yi-Chun Kuo ₃ , Shuguang Wei ₄ , Anwu Zhou ₄ , Zhongju Zou _{1,

2} , Jenna Yehl ₅ , Matthew J. Ranaghan ₅ , Adam Skepner ₅ , Joshua A. Bittker ₅ , Jose R. Perez ₅ , Bruce A. Posner ₄ , Beth Levine _{1,

2,

6}

Affiliation

Autophagy, a lysosomal degradation pathway, plays a crucial role in cellular homeostasis, development, immunity, tumor suppression, metabolism, prevention of neurodegeneration, and lifespan extension. Thus, pharmacological stimulation of autophagy may be an effective approach for preventing or treating certain human diseases and/or aging. We sought to establish a method for developing new chemical compounds that specifically induce autophagy. To do this, we developed two assays to identify compounds that target a key regulatory node of autophagy induction—specifically, the binding of Bcl-2 (a negative regulator of autophagy) to Beclin 1 (an allosteric modulator of the Beclin 1/VPS34 lipid kinase complex that functions in autophagy initiation). These assays use either a split-luciferase assay to measure Beclin 1/Bcl-2 binding in cells or an AlphaLISA assay to directly measure direct Beclin 1/Bcl-2 binding in vitro. We screened two different chemical compound libraries, comprising ∼300 K compounds, to identify small molecules that disrupt Beclin 1/Bcl-2 binding and induce autophagy. Three novel compounds were identified that directly inhibit Beclin 1/Bcl-2 interaction with an IC₅₀ in the micromolar range and increase autophagic flux. These compounds do not demonstrate significant cytotoxicity, and they exert selectivity for disruption of Bcl-2 binding to the BH3 domain of Beclin 1 compared with the BH3 domain of the pro-apoptotic Bcl-2 family members, Bax and Bim. Thus, we have identified candidate molecules that serve as lead templates for developing potent and selective Beclin 1/Bcl-2 inhibitors that may be clinically useful as autophagy-inducing agents.

中文翻译：

高通量筛选，以鉴定通过破坏Beclin 1 / Bcl-2结合起作用的自噬诱导物。

自噬是一种溶酶体降解途径，在细胞稳态，发育，免疫，肿瘤抑制，新陈代谢，神经变性的预防和寿命延长中起着至关重要的作用。因此，自噬的药理刺激可能是预防或治疗某些人类疾病和/或衰老的有效方法。我们试图建立一种方法来开发特异性诱导自噬的新化合物。为此，我们开发了两种检测方法来鉴定靶向自噬诱导关键调控节点的化合物，即Bcl-2（自噬的负调节剂）与Beclin 1（Beclin 1 / VPS34脂质的变构调节剂）结合在自噬启动中起作用的激酶复合物）。体外。我们筛选了包含〜300 K化合物的两个不同的化合物文库，以鉴定破坏Beclin 1 / Bcl-2结合并诱导自噬的小分子。鉴定了三种新化合物，它们直接抑制Beclin 1 / Bcl-2与IC _50的相互作用在微摩尔范围内并增加自噬通量。这些化合物没有显示出明显的细胞毒性，与凋亡前Bcl-2家族成员Bax和Bim的BH3结构域相比，它们对破坏Beclin 1的BH3结构域的Bcl-2结合具有选择性。因此，我们已经确定了候选分子，它们可以用作开发有效和选择性的Beclin 1 / Bcl-2抑制剂的先导模板，这些抑制剂在临床上可作为自噬诱导剂。

更新日期：2018-06-07

点击分享查看原文

点击收藏

公开下载

阅读更多本刊最新论文本刊介绍/投稿指南11