Purpose

5′ adenosine monophosphate-activated kinase (AMPK) is an essential regulator of cellular energy homeostasis and has been associated with different pathologies, including cancer. Precisely defining the biological role of AMPK necessitates the availability of a potent and selective inhibitor.

Methods

High-throughput screening and chemical optimization were performed to identify a novel AMPK inhibitor. Cell proliferation and mechanistic assays, as well as gene expression analysis and chromatin immunoprecipitation were used to investigate the cellular impact as well as the crosstalk between lipid metabolism and androgen signaling in prostate cancer models. Also, fatty acid turnover was determined by examining lipid droplet formation.

Results

We identified BAY-3827 as a novel and potent AMPK inhibitor with additional activity against ribosomal 6 kinase (RSK) family members. It displays strong anti-proliferative effects in androgen-dependent prostate cancer cell lines. Analysis of genes involved in AMPK signaling revealed that the expression of those encoding 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGCR), fatty acid synthase (FASN) and 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 2 (PFKFB2), all of which are involved in lipid metabolism, was strongly upregulated by androgen in responsive models. Chromatin immunoprecipitation DNA-sequencing (ChIP-seq) analysis identified several androgen receptor (AR) binding peaks in the HMGCR and PFKFB2 genes. BAY-3827 strongly down-regulated the expression of lipase E (LIPE), cAMP-dependent protein kinase type II-beta regulatory subunit (PRKAR2B) and serine-threonine kinase AKT3 in responsive prostate cancer cell lines. Also, the expression of members of the carnitine palmitoyl-transferase 1 (CPT1) family was inhibited by BAY-3827, and this was paralleled by impaired lipid flux.

Conclusions

The availability of the potent inhibitor BAY-3827 will contribute to a better understanding of the role of AMPK signaling in cancer, especially in prostate cancer.

中文翻译：

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有效的 AMPK 抑制剂 BAY-3827 在雄激素依赖性前列腺癌模型中显示出强大的功效

目的

5' 腺苷单磷酸活化激酶 (AMPK) 是细胞能量稳态的重要调节剂，并与包括癌症在内的不同病理有关。准确定义 AMPK 的生物学作用需要有效和选择性抑制剂的可用性。

方法

进行高通量筛选和化学优化以鉴定新型 AMPK 抑制剂。细胞增殖和机制测定，以及基因表达分析和染色质免疫沉淀用于研究前列腺癌模型中的细胞影响以及脂质代谢和雄激素信号传导之间的串扰。此外，通过检查脂滴形成来确定脂肪酸周转率。

结果

我们将 BAY-3827 鉴定为一种新型有效的 AMPK 抑制剂，对核糖体 6 激酶 (RSK) 家族成员具有额外的活性。它在雄激素依赖性前列腺癌细胞系中显示出强大的抗增殖作用。对参与 AMPK 信号传导的基因的分析表明，编码 3-羟基-3-甲基-戊二酰辅酶 A 还原酶 (HMGCR)、脂肪酸合酶 (FASN) 和 6-磷酸果糖-2-激酶/果糖-2 的那些基因的表达， 6-双磷酸酶 2 (PFKFB2)，所有这些都参与脂质代谢，在响应模型中被雄激素强烈上调。染色质免疫沉淀 DNA 测序 (ChIP-seq) 分析确定了 HMGCR 和 PFKFB2 基因中的几个雄激素受体 (AR) 结合峰。BAY-3827 强烈下调脂肪酶 E (LIPE) 的表达，反应性前列腺癌细胞系中的 cAMP 依赖性蛋白激酶 II-β 调节亚基 (PRKAR2B) 和丝氨酸-苏氨酸激酶 AKT3。此外，肉碱棕榈酰转移酶 1 (CPT1) 家族成员的表达受到 BAY-3827 的抑制，同时脂质通量受损。

结论

有效抑制剂 BAY-3827 的可用性将有助于更好地了解 AMPK 信号在癌症中的作用，尤其是在前列腺癌中。