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ACS Chem Biol DOI:10.1021/acschembio.8b00421

High-Throughput Screens To Identify Autophagy Inducers That Function by Disrupting Beclin 1/Bcl-2 Binding.

Publication TypeJournal Article
Year of Publication2018
AuthorsChiang, W-C, Wei, Y, Kuo, Y-C, Wei, S, Zhou, A, Zou, Z, Yehl, J, Ranaghan, MJ, Skepner, A, Bittker, JA, Perez, JR, Posner, BA, Levine, B
JournalACS Chem Biol
Volume13
Issue8
Pages2247-2260
Date Published2018 08 17
ISSN1554-8937
KeywordsAutophagy, Beclin-1, Cell Survival, Drug Evaluation, Preclinical, HeLa Cells, High-Throughput Screening Assays, Humans, Protein Binding, Protein Interaction Maps, Proto-Oncogene Proteins c-bcl-2
Abstract

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 IC 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.

DOI10.1021/acschembio.8b00421
Pubmed

http://www.ncbi.nlm.nih.gov/pubmed/29878747?dopt=Abstract

Alternate JournalACS Chem. Biol.
PubMed ID29878747
PubMed Central IDPMC6250051
Grant ListP30 CA142543 / CA / NCI NIH HHS / United States
R01 CA109618 / CA / NCI NIH HHS / United States
U19 AI109725 / AI / NIAID NIH HHS / United States