We identify the Plasmodium falciparum acetyl-coenzyme A synthetase (PfAcAS) as a druggable target, using genetic and chemical validation. In vitro evolution of resistance with two antiplasmodial drug-like compounds (MMV019721 and MMV084978) selects for mutations in PfAcAS. Metabolic profiling of compound-treated parasites reveals changes in acetyl-CoA levels for both compounds. Genome editing confirms that mutations in PfAcAS are sufficient to confer resistance. Knockdown studies demonstrate that PfAcAS is essential for asexual growth, and partial knockdown induces hypersensitivity to both compounds. In vitro biochemical assays using recombinantly expressed PfAcAS validates that MMV019721 and MMV084978 directly inhibit the enzyme by preventing CoA and acetate binding, respectively. Immunolocalization studies reveal that PfAcAS is primarily localized to the nucleus. Functional studies demonstrate inhibition of histone acetylation in compound-treated wild-type, but not in resistant parasites. Our findings identify and validate PfAcAS as an essential, druggable target involved in the epigenetic regulation of gene expression.

我们使用遗传和化学验证将恶性疟原虫乙酰辅酶 A 合成酶 ( Pf AcAS )确定为药物靶点。两种抗疟原虫类药物化合物（MMV019721 和 MMV084978）的耐药性体外进化选择了Pf AcAS 中的突变。化合物处理的寄生虫的代谢分析揭示了两种化合物的乙酰辅酶 A 水平的变化。基因组编辑证实Pf AcAS 中的突变足以产生抗性。击倒研究表明Pf AcAS 对无性生长至关重要，部分击倒会导致对这两种化合物的超敏反应。体外使用重组表达的Pf AcAS 的生化测定证实 MMV019721 和 MMV084978 分别通过阻止 CoA 和乙酸结合来直接抑制酶。免疫定位研究表明Pf AcAS 主要定位于细胞核。功能研究表明在化合物处理的野生型中抑制了组蛋白乙酰化，但在抗性寄生虫中没有。我们的研究结果确定并验证了Pf AcAS 作为参与基因表达表观遗传调控的重要药物靶点。