胰腺导管腺癌（PDAC）是一种高度转移的恶性肿瘤，目前尚无有效的抗转移疗法。在此，我们采用单细胞RNA测序和代谢组学分析来证明转移细胞高度表达粘着斑激酶（FAK），FAK通过重塑胆碱激酶α（CHKα）依赖性胆碱代谢来促进转移。我们设计了一种新型CHKα抑制剂CHKI-03，并在多个临床前模型中验证了其抑制转移的功效。经典和新合成的小分子抑制剂之前已被用来评估在各种动物模型中靶向 mTOR 和 CHKα 的治疗潜力。机制上，FAK激活mTOR及其下游HIF-1α，从而升高CHKα表达，促进PDAC细胞的增殖、迁移和侵袭，以及肿瘤的生长和转移。一致地，FAK 和 CHKα 的高表达水平与 PDAC 患者的不良预后相关。值得注意的是，CHK1-03 抑制 CHKα 表达，还抑制 mTORC1 磷酸化，破坏 mTORC1-CHKα 正反馈环。此外，CHKI-03与mTORC1抑制剂雷帕霉素联合使用可协同抑制PDX模型中的肿瘤生长和转移。 CHKI-03 和雷帕霉素的组合在吉西他滨耐药的 PDO 模型中表现出相当大的治疗效果。我们的研究结果揭示了 mTORC1-CHKα 环依赖性胆碱代谢重编程调节 PDAC 转移的关键机制，突出了这种新疗法治疗 PDAC 转移的治疗潜力。版权所有 © 2024 Elsevier B.V. 保留所有权利。

Pancreatic ductal adenocarcinoma (PDAC) is a highly metastatic malignancy for which there are currently no effective anti-metastatic therapies. Herein, we employed single-cell RNA sequencing and metabolomics analysis to demonstrate that metastatic cells highly express focal adhesion kinase (FAK), which promotes metastasis by remodeling choline kinase α (CHKα)-dependent choline metabolism. We designed a novel CHKα inhibitor, CHKI-03, and verified its efficacy in inhibiting metastasis in multiple preclinical models. Classical and newly synthesized small-molecule inhibitors have previously been used to assess the therapeutic potential of targeting mTOR and CHKα in various animal models. Mechanistically, FAK activated mTOR and its downstream HIF-1α, thereby elevating CHKα expression and promoting the proliferation, migration, and invasion of PDAC cells, as well as tumor growth and metastasis. Consistently, high expression levels of both FAK and CHKα are correlated with poor prognosis in patients with PDAC. Notably, CHK1-03 inhibited CHKα expression and also suppressed mTORC1 phosphorylation, disrupting the mTORC1-CHKα positive feedback loop. In addition, the combination of CHKI-03 and the mTORC1 inhibitor rapamycin synergistically inhibited tumor growth and metastasis in PDX models. The combination of CHKI-03 and rapamycin demonstrates considerable therapeutic efficacy in PDO models resistant to gemcitabine. Our findings reveal a pivotal mechanism underlying PDAC metastasis regulated by mTORC1-CHKα loop-dependent choline metabolism reprogramming, highlighting the therapeutic potential of this novel regimen for treating PDAC metastasis.Copyright © 2024 Elsevier B.V. All rights reserved.