CAD manipulates tumor intrinsic DHO/UBE4B/NF-κB pathway and fuels macrophage cross-talk, promoting HCC metastasis
Jiaomeng Pan, Mao Zhang, Dongning Rao, Junjie Ma, Xia Shen, Haokai Qin, Kun Gan, Jianhua Lin, Yingying Huang, Chen Sang, Juan Zhang, Jiaqiang Ma, Yingcheng Wu, Zheng Tang, Daming Gao, Qiang Gao, Liu‐Xiao Yang, Jia Fan
Abstract
BACKGROUND AND AIMS: Portal vein tumor thrombosis (PVTT), an indicator of clinical metastasis, significantly shortens the lifespan of patients with HCC, and no effective treatment has been established. We aimed to illustrate the mechanisms underlying PVTT formation and tumor metastasis and identify potential targets for clinical intervention. APPROACH AND RESULTS: Multi-omics data of 159 patients with HCC (including 37 cases with PVTT) was analyzed to identify contributors to PVTT formation and tumor metastasis. In vitro and in vivo experiments were performed to confirm the critical role of carbamoyl-phosphate synthetase 2, aspartate transcarbamylase, and dihydroorotase (CAD) in HCC metastasis. Metabolomics and transcriptomics techniques, single-cell RNA sequencing, combined with experimental verification, were complemented to illustrate mechanisms underlying CAD-induced pro-metastatic efficacy. Analysis of proteogenomic data of HCC cohort identified CAD as the key contributor to PVTT formation and tumor metastasis in HCC. Further experiments confirmed that high CAD expression could significantly promote HCC metastasis, and vice versa. Mechanistically, CAD manipulated de novo pyrimidine anabolism, leading to dihydroorotic acid (DHO) accumulation, which is directly bound to ubiquitination factor E4B (UBE4B). UBE4B subsequently regulated JAK1 ubiquitination and activated the NF-κB pathway to promote epithelial-mesenchymal transition of HCC cells. In addition, CAD generated an immunosuppressive milieu conducive to HCC metastasis by recruiting and reprogramming macrophages into a "pro-tumor" phenotype. Consequently, the metastatic capability of HCC was remarkably enhanced. CONCLUSIONS: Therapy targeting CAD may offer a promising approach to curb HCC metastasis by reducing tumor cells' metastatic potential and also shifting the tumor microenvironment toward a less pro-metastatic state.