Page 3 - I. Chemical biology and drug discovery

Target Identification of a natural compound that ameliorates
atherosclerosis via autophagy-inducing activity

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Minjeong Ko , Dongjin Lee , Ju Yeon Lee , Jin Young Kim , Jong Shin Yoo and Ho Jeong Kwon 1,*
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1 Chemical Genomics Global Research Laboratory, Department of Biotechnology, College of Life Science & Biotechnology, Yonsei University, Seoul 120-749, Korea.
2 Korea Basic Science Institute, Cheongwon-Gun, Chungbuk 363-883, Korea. * Corresponding author: kwonhj@yonsei.ac.kr
BACKGROUND AIM
• Atherosclerosis is a life-threatening chronic inflammatory disease in which • This study provides new insights into the mechanism of an anti-atherosclerotic
natural compound in linking with autophagy.
blood vessels are blocked due to the accumulation of cholesterol and
plaque formation. • To uncover the mode of action of compound YCGX, a combination of drug affinity
• Autophagy in endothelial cell is involved in vascular lipid homeostasis responsive target stability (DARTS) and LC-MS/MS method was applied to identify the
target protein of compound YCGX. The target protein candidate was validated by
which can protect the atherosclerotic lesion formation. However, the DARTS western blot analysis.
deficiency of endothelial autophagy and malfunctions of macrophage • Further validation of interaction of compound with target protein was conducted with
autophagy trigger the atherosclerotic plaque formation. knockdown of the target gene resulting in increase of the autophagic features.
METHODS

Target identification using ‘DARTS (Drug Affinity Responsive Target Stability)-LC-MS/MS’
• DARTS is a new technology for protein target identification of non-labeled small molecules.
• Target protein bound with drug of interest has high stability against pronase treatment and
pronase can not easily degrade drug-bound proteins.

RESULTS

Figure 1 Figure 2 Figure 4
DMSO YCGX (5 µM)
YCGX
NT 24 48 72 24 48 72 (h)
p62
LC3B Ⅰ
Ⅱ
β-actin

Figure 3 Summary

Figure1. Compound YCGX induced autophagic flux in HUVECs
Figure 2. Compound YCGX inhibited foam cell formation via increasing lysosomal activities in RAW 264.7 macrophages
Figure 3. Target ID & VD through DARTS-LC-MS/MS
Figure 4. Is target protein H responsible for autophagy activity?
CONCLUSION REFERENCES ACKNOWLEDGEMENTS

• Compound YCGX induces autophagy in Supported by BK21 PLUS, NRF (MSIP;
HUVECs. • Lomenick et al. Target identification using drug 2015K1A1A2028365, 2018M3A9C4076477)
• Compound YCGX inhibited foam cell formation affinity responsive target stability (DARTS). Curr
Protoc Chem Biol. 2011
in oxLDL-induced RAW 264.7 macrophages via • Sergin et al. Exploiting macrophage autophagy-
inducing lysosome activity. lysosomal biogenesis as a therapy for atherosclerosis. Contact information
• Target proteins of compound YCGX is Nat. Commun. 2017
identified by DARTS LC-MS/MS and validated • Guo et al. The role of the LncRNA-FA2H-2-MLKL
via biological analysis. pathway in atherosclerosis by regulation of autophagy • Presenter : Minjeong Ko
komj0714@yonsei.ac.kr
• Target validation for protein H, the final target flux and inflammation through mTOR-dependent • Correspondence : Ho Jeong Kwon
protein of compound YCGX is underway. signaling. Cell Death Differ. 2019 kwonhj@yonsei.ac.kr