最近，铜及其转运蛋白铜转运蛋白1（CTR1）在肿瘤发生中扮演主导角色的研究得到了探索。然而，CTR1的上游调控和铜螯合剂在恶性肿瘤中的联合干预仍然不清楚。我们采用CRISPR/Cas9基因组筛查方法，鉴定了CTR1上游激酶；采用免疫荧光检测CTR1定位；进行体外激酶酶解和质谱实验探测CTR1的磷酸化；进行泛素化实验验证CTR1的稳定性；进行集落形成、EdU标记、Annexin V-FITC/PI基于凋亡实验来检测药物对细胞生长和凋亡的影响；并使用异种移植小鼠模型来研究药物在体内的作用。 我们发现，CTR1经历AMPK介导的磷酸化作用，促进CTR1的稳定和膜转运，并通过影响Nedd4l的相互作用，从而增加CTR1在乳腺癌中的致癌作用。重要的是，AMPK的激动剂丢陟羧酸可显著增强CTR1水平，并导致AMPK激动剂和铜螯合剂联合治疗乳腺癌。 我们的发现不仅揭示了通过AMPK介导CTR1磷酸化和稳定性来实现能量响应和铜摄取的交互作用，而且还强调了通过AMPK激动剂和铜螯合剂的联合使用来对抗乳腺癌的策略。 版权所有©2022。作者（们）在Springer Nature Limited独家许可下发表。

Predominant roles of copper and its transporter, copper transporter 1 (CTR1), in tumorigenesis have been explored recently; however, the upstream regulation of CTR1 and combinational intervention of copper chelators in malignancies remain largely unclear.CRISPR/Cas9-based kinome screening was used to identify the CTR1 upstream kinases. Immunofluorescence assays were utilised to detect CTR1 localisation. In vitro kinase assays and mass spectrometry were performed to detect CTR1 phosphorylation. Ubiquitination assays were performed to validate CTR1 stability. Colony formation, EdU labelling, Annexin V-FITC/PI-based apoptosis assays were carried out to detect the drug effect on cell growth and apoptosis. Xenografted mouse models were employed to investigate drug effects in vivo.We identify that CTR1 undergoes AMPK-mediated phosphorylation, which enhances CTR1 stabilisation and membrane translocation by affecting Nedd4l interaction, resulting in increased oncogenic roles in breast cancer. Importantly, activation of AMPK with its agonist metformin markedly enhances CTR1 levels, and leads to the combinational usage of AMPK agonists and copper chelators for breast cancer treatment.Our findings not only reveal the crosstalk between energy response and copper uptake via AMPK-mediated CTR1 phosphorylation and stability but also highlight the strategy to combat breast cancer by a combination of AMPK agonists and copper chelators.The connection between energy response and copper homoeostasis is linked by AMPK phosphorylating and stabilising CTR1, which provides a promising strategy to combat breast cancer by combining AMPK agonists and copper chelators.© 2022. The Author(s), under exclusive licence to Springer Nature Limited.