Drug evaluation based on phosphomimetic PDHA1 reveals the complexity of activity-related cell death in A549 non-small cell lung cancer cells
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Cancer cells predominantly generate energy via glycolysis, evenin the presence of oxygen, to support abnormal cell proliferation. Suppression of PDHA1 by PDK1 prevents the conversionof cytoplasmic pyruvate into Acetyl-CoA. Several PDK inhibitorshave been identified, but their clinical applications havenot been successful for unclear reasons. In this study, endogenousPDHA1 in A549 cells was silenced by the CRISPR/Cas9system, and PDHA1WT and PDHA13SD were transduced. SincePDHA13SD cannot be phosphorylated by PDKs, it was used toevaluate the specific activity of PDK inhibitors. This study highlightsthat PDHA1WT and PDHA13SD A549 cells can be used asa cell-based PDK inhibitor-distinction system to examine therelationship between PDH activity and cell death by establishedPDK inhibitors. Leelamine, huzhangoside A and otobaphenolinduced PDH activity-dependent apoptosis, whereas AZD7545,VER-246608 and DCA effectively enhanced PDHA1 activitybut little toxic to cancer cells. Furthermore, the activity of phosphomimeticPDHA1 revealed the complexity of its regulation,which requires further in-depth investigation.
Abstract
Cancer cells predominantly generate energy via glycolysis, evenin the presence of oxygen, to support abnormal cell proliferation. Suppression of PDHA1 by PDK1 prevents the conversionof cytoplasmic pyruvate into Acetyl-CoA. Several PDK inhibitorshave been identified, but their clinical applications havenot been successful for unclear reasons. In this study, endogenousPDHA1 in A549 cells was silenced by the CRISPR/Cas9system, and PDHA1WT and PDHA13SD were transduced. SincePDHA13SD cannot be phosphorylated by PDKs, it was used toevaluate the specific activity of PDK inhibitors. This study highlightsthat PDHA1WT and PDHA13SD A549 cells can be used asa cell-based PDK inhibitor-distinction system to examine therelationship between PDH activity and cell death by establishedPDK inhibitors. Leelamine, huzhangoside A and otobaphenolinduced PDH activity-dependent apoptosis, whereas AZD7545,VER-246608 and DCA effectively enhanced PDHA1 activitybut little toxic to cancer cells. Furthermore, the activity of phosphomimeticPDHA1 revealed the complexity of its regulation,which requires further in-depth investigation.
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