清大生醫學院生科系王雯靜教授（左一）與研究團隊成員

Professor Wen-Ching Wang of Department of Life Science, National Tsing Hua University, with research team member.

論文標題Paper Title

GSH as an A-A Type Allosteric Activator of PKM2: Modulating Cancer Cell Homeostasis and Ferroptosis Susceptibility

學術期刊刊名Journal Name

先進科學Advanced Science

研究團隊 Research Team

主持人Principle Investigator：

王雯靜 教授  Wen-Ching Wang, Professor

國立清華大學生命科學系/分子與細胞生物研究所

Department of Life Science

Institute of Molecular and Cellular Biology

 National Tsing Hua University (NTHU).

參與者Participants:：

陳粲然 博士 Tsan-Jan Chen

國立清華大學分子與細胞生物研究所

Institute of Molecular and Cellular Biology, National Tsing Hua University (NTHU).

龔行健 講座教授Hsing-Jien Kung, Chair Professor

台北醫學大學/UC Davis團隊

Taipei Medical University (TMU)/ University of California, Davis(UC Davis)

楊慕華 副校長Muh-Hwa Yang, Vice President

國立陽明交通大學

National Yang Ming Chiao Tung University(NYCU)

林峻宇 副教授Chun-Yu Lin, Associate Professor

國立陽明交通大學

National Yang Ming Chiao Tung University(NYCU)

鄭美玲 教授Mei-Ling Cheng, Professor

長庚大學

Chang Gung University(CGU)

駱啟仁 博士 Chi-Jen Lo

長庚大學

Chang Gung University(CGU)

平岡泰 Yasushi Hiraoka

大阪大學

Osaka University

原口德子 Tokuko Haraguchi

大阪大學

Osaka University

摘要

Abstract

本研究鑑定出穀胱甘肽（GSH）為丙酮酸激酶 M2（PKM2）的一種內源性 A-A 型變構活化劑，透過在 A-A 介面結合，將其穩定於活性的四聚體構型。此 PKM2–GSH 的交互作用將 GSH 代謝與鐵死亡的調控連結起來。跨癌別的轉錄體分析顯示，GSH、SLC7A11、PKM2、糖解作用與鐵死亡途徑之間具有高度相關性。透過耗竭 GSH 並活化 PKM2，我們在依賴 PKM2 的癌症模型中增強了鐵死亡。此策略導致中心碳代謝與脂質代謝的顯著改變，破壞粒線體功能，並誘發鐵死亡型細胞死亡。在異種移植模型中，合併處理可顯著抑制腫瘤生長。較高的 PKM2 與 SLC7A11 表現量與較差的存活預後相關，顯示其作為鐵死亡導向治療生物標記的潛力。我們的研究結果揭示了 GSH 在細胞穩態中的雙重角色，並指出 PKM2–GSH–SLC7A11 軸為侵襲性癌症的潛在治療標的。

This study identifies glutathione (GSH) as an endogenous A-A type allosteric activator of pyruvate kinase M2 (PKM2), stabilizing it in its active tetrameric form through binding at the A-A interface. This PKM2-GSH interaction links GSH metabolism to ferroptosis regulation. Transcriptomic analyses across cancers demonstrate strong correlations between GSH, SLC7A11, PKM2, glycolysis, and ferroptosis pathways. By depleting GSH and activating PKM2, we enhance ferroptosis in PKM2-dependent cancer models. This approach leads to significant changes in central carbon and lipid metabolism, disrupts mitochondrial function, and drives ferroptotic cell death. The combined treatment markedly suppresses tumor growth in a xenograft model. Elevated PKM2 and SLC7A11 expression levels correlate with poorer survival outcomes, indicating their potential as biomarkers for ferroptosis-based therapy. Our findings demonstrate a dual role for GSH in cellular homeostasis and identify the PKM2-GSH-SLC7A11 axis as a therapeutic target for aggressive cancers.

研究成果Result/Contributions

PKM2 受到精細且複雜的變構調控，並且是癌症代謝的核心驅動因子。腫瘤細胞利用 PKM2 重新編程葡萄糖的利用方式，將碳流由三羧酸循環轉向乳酸生成與戊糖磷酸途徑—這種被稱為瓦氏效應（Warburg effect）的代謝適應，有助於支持細胞的快速增殖。

在過去幾年中，我們發現 PKM2 透過與 GSH 的交互作用，在調控鐵死亡型細胞死亡中扮演一個先前未被重視的角色，並建立了中心碳代謝與脂質活性氧（ROS）恆定之間的機制性連結。我們鑑定出 GSH 為一種先前未被發現的異位活化劑，能提升 PKM2 的酵素活性並穩定其活性四聚體構型。對 PKM2–GSH 複合體的結構分析顯示，GSH 佔據了位於寡聚化介面的一個結合位點。值得注意的是，此區域與小分子 PKM2 活化劑 DASA-58 的結合位點重疊，顯示兩者可能具有共同的異構調控機制。

利用 erastin（SLC7A11 的抑制劑）進行藥理性方式耗竭細胞內 GSH，會降低 PKM2 的活性並在 SAS 細胞中誘發鐵死亡。重要的是，在 GSH 被耗竭的條件下合併使用 DASA-58，會進一步增強鐵死亡效應。此細胞死亡表型可被鐵死亡抑制劑特異性挽救，但不會被針對細胞凋亡或壞死性凋亡的抑制劑所影響。此外，在小鼠異種移植模型中，DASA-58 可增強 erastin 的腫瘤抑制效果。

綜合而言，這些發現揭示了一條先前未被認識的 PKM2–GSH 軸線參與鐵死亡調控，並指出 PKM2 與 SLC7A11 為具潛力的癌症生物標記與治療標的。

PKM2 undergoes intricate allosteric regulation and is a central driver of cancer metabolism. Tumor cells exploit PKM2 to rewire glucose utilization, channeling carbon away from the tricarboxylic acid cycle toward lactate production and the pentose phosphate pathway—an adaptation known as the Warburg effect that supports rapid proliferation.

Over the past year, we have uncovered an unappreciated role for PKM2 in controlling ferroptotic cell death through its interaction with GSH, establishing a mechanistic link between central carbon metabolism and lipid ROS homeostasis. We identified GSH as a previously unrecognized allosteric activator that boosts PKM2 enzymatic activity and stabilizes its active tetrameric state. Structural analysis of the PKM2–GSH complex reveals that GSH occupies a pocket at the oligomerization interface. Notably, this region overlaps with the binding site of DASA-58, a small-molecule PKM2 activator, indicating a shared allosteric mechanism.

Pharmacological depletion of intracellular GSH using erastin, an inhibitor of SLC7A11, diminishes PKM2 activity and triggers ferroptotic cell death in SAS cells. Importantly, co-treatment with DASA-58 under GSH-depleted conditions further amplifies ferroptosis. This phenotype is specifically rescued by ferroptosis inhibitors, but not by inhibitors targeting apoptosis or necroptosis. Moreover, in mouse xenograft models, DASA-58 enhances the tumor-suppressive effects of erastin.

Together, these findings define a previously unrecognized PKM2–GSH axis in ferroptosis regulation and position PKM2 and SLC7A11 as promising biomarkers and therapeutic targets in cancer.

論文連結 Paper Link

https://advanced.onlinelibrary.wiley.com/doi/full/10.1002/advs.202519368

自由健康網：https://health.ltn.com.tw/article/breakingnews/5286902

中時新聞網：https://www.chinatimes.com/realtimenews/20251223003015-260418?chdtv

聯合新聞網：https://udn.com/news/story/6928/9220661?from=searchresult

中央社：https://www.cna.com.tw/news/ahel/202512230101.aspx

台灣好新聞：https://reurl.cc/bNeoQ6

大紀元：https://www.epochtimes.com/b5/25/12/23/n14660882.htm

科技新報：https://technews.tw/2025/12/23/tsinghua-university-research-breaks-through-the-weaknesses-of-cancer-cells/

福爾摩沙網：https://formosalive.com/archives/230439

好報新聞網：https://www.hbnews.com.tw/Report/News/e0107b032s07

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