Page 62 - ebook

[A. Biochemistry/Molecular Biology] A-31

Drug evaluation based on phosphomimetic PDHA1 reveals

the complexity of activity–related cell death in A549 non-

small cell lung cancer cells

Ling Jin¹,², Minkyoung Cho², Sung-Jin Bae², Ki-Tae Ha¹,²*

¹Department of Korean Medical Science, School of Korean Medicine, Pusan National University, Yangsan 50612,

Korea, ²Korean Medical Research Center for Healthy Aging, Pusan National University, Yangsan 50612, Korea

Cancer cells predominantly generate energy via glycolysis, even in the presence of oxygen, to support abnormal

cell proliferation. Suppression of PDHA1 by PDK1 prevents the conversion of cytoplasmic pyruvate into Acetyl-CoA.
Several PDK inhibitors have been identified, but their clinical applications have not been successful for unclear

reasons. In this study, endogenous PDHA1 in A549 cells was silenced by the CRISPR/Cas9 system, and PDHA1WT
and PDHA13SD were transduced. Since PDHA13SD cannot be phosphorylated by PDKs, it was used to evaluate the

specific activity of PDK inhibitors. This study highlights that PDHA1WT and PDHA13SD A549 cells can be used as a
cell-based PDK inhibitor–distinction system to examine the relationship between PDH activity and cell death by

established PDK inhibitors. Leelamine, huzhangoside A and otobaphenol induced PDH activity-dependent apoptosis,
whereas AZD7545, VER-246608 and DCA effectively enhanced PDHA1 activity but little toxic to cancer cells.
Furthermore, the activity of phosphomimetic PDHA1 revealed the complexity of its regulation, which requires further

in-depth investigation.