Treatment of acute leukemia is challenging due to genetic heterogeneity between and even within patients. Leukemic stem cells (LSCs) are relatively drug-resistant and frequently lead to relapse. Their plasticity and capacity to adapt to extracellular stress, in which mitochondrial metabolism and autophagy play important roles, further complicates treatment. Genetic models of phosphatidylinositol-5-phosphate 4-kinase type 2 proteins (PIP4K2s) inhibition demonstrated the relevance of these enzymes in mitochondrial homeostasis and autophagic flux. Here, we uncover the cellular and molecular effects of THZ-P1-2, a pan-inhibitor of PIP4K2s, in acute leukemia cells. THZ-P1-2 reduced cell viability and induced DNA damage, apoptosis, loss of mitochondrial membrane potential, and accumulation of acidic vesicular organelles. Protein expression analysis revealed that THZ-P1-2 impaired autophagy flux. In addition, THZ-P1-2 induced cell differentiation and showed synergistic effects with venetoclax in resistant leukemic models. In primary leukemia cells, LC-MS/MS-based proteome analysis revealed that sensitivity to THZ-P1-2 was associated with mitochondrial metabolism, cell cycle, cell-of-origin, and the TP53 pathway. Minimal effects of THZ-P1-2 observed in healthy CD34⁺ cells suggested a favorable therapeutic window. Our study provides insight into pharmacological inhibition of PIP4Ks targeting mitochondrial homeostasis and autophagy shedding light on a new class of drugs for acute leukemias.

中文翻译：

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PIP4K2 抑制剂 THZ-P1-2 通过破坏线粒体稳态和自噬表现出抗白血病活性

由于患者之间甚至患者内部的遗传异质性，急性白血病的治疗具有挑战性。白血病干细胞 (LSCs) 具有相对的耐药性并经常导致复发。它们的可塑性和适应细胞外压力的能力，其中线粒体代谢和自噬发挥重要作用，使治疗进一步复杂化。磷脂酰肌醇-5-磷酸 4-激酶 2 型蛋白 (PIP4K2s) 抑制的遗传模型证明了这些酶在线粒体稳态和自噬通量中的相关性。在这里，我们揭示了 PIP4K2 的泛抑制剂 THZ-P1-2 在急性白血病细胞中的细胞和分子效应。THZ-P1-2 降低细胞活力并诱导 DNA 损伤、细胞凋亡、线粒体膜电位丧失和酸性水泡细胞器的积累。蛋白质表达分析显示 THZ-P1-2 削弱了自噬通量。此外，THZ-P1-2 诱导细胞分化并在耐药白血病模型中显示出与 venetoclax 的协同作用。在原发性白血病细胞中，基于 LC-MS/MS 的蛋白质组分析显示，对 THZ-P1-2 的敏感性与线粒体代谢、细胞周期、细胞来源和 TP53 通路有关。在健康 CD34 中观察到 THZ-P1-2 的最小影响⁺细胞表明有利的治疗窗口。我们的研究提供了对靶向线粒体稳态和自噬的 PIP4Ks 的药理学抑制作用的见解，从而揭示了一类新的急性白血病药物。