Wang Menghui, Li Yueheng, Su Junhui, Dong Xinjue, Liu Ao, Yang Yuqi, Tang Xinyi, Chen Ruijie, Li QingQuan, Wang Hongshan, Xiao Hong
Department of Pharmacology, School of Pharmacy, Fudan University, Shanghai, 201203, P. R. China.
Department of Pancreatic Surgery, Zhongshan Hospital, Fudan University, Shanghai, P. R. China.
J Transl Med. 2025 Apr 11;23(1):435. doi: 10.1186/s12967-025-06333-5.
Many cases of advanced hepatocellular carcinoma (HCC) are resistant to the widely used drug sorafenib, which worsens prognosis. While many studies have explored how acquired resistance emerges during drug exposure, the mechanism underlying primary resistance before treatment still remain elusive.
Single-cell lineage tracing and RNA sequencing were performed to identify primary sorafenib-resistant lineages in HCC. Differential gene expression analysis was employed to identify the biomarkers of drug-resistant lineage cells. Cell viability and colony formation assays were adopted to assess the involvement of S100A14 on sorafenib resistance. Co-immunoprecipitation (CO-IP) and mass spectrometry analysis were conducted to uncover the downstream targets and regulatory mechanisms of S100A14 in primary resistance to sorafenib. In vivo mouse xenograft experiments were carried out to assess the effect of S100A14 or its interacting protein glutaminase (GLS) on primary resistance to sorafenib in HCC.
Single-cell lineage tracing identified a cluster of sorafenib primary resistant cells, and S100A14, a Ca-binding protein, was determined to be a critical biomarker for primary resistance to sorafenib. Knockdown of S100A14 significantly increases sorafenib treatment sensitivity in HCC cells. Mechanistically, S100A14 binds to GLS and blocks its phosphorylation at residues Y308 and S314, which in turn inhibits its ubiquitination and subsequent degradation. By stabilizing GLS, S100A14 reduces oxidative stress in HCC cells, thereby antagonizing sorafenib-induced apoptosis. Inhibiting S100A14 or GLS significantly improved sorafenib efficacy against xenograft tumors in vivo.
Our results demonstrate that S100A14 plays a pivotal role in promoting primary resistance to sorafenib by stabilizing GLS to reduce oxidative stress, and acts as a potential therapeutic target to enhance the efficacy of sorafenib in HCC patients.
许多晚期肝细胞癌（HCC）病例对广泛使用的药物索拉非尼耐药，这会使预后恶化。虽然许多研究探讨了在药物暴露期间获得性耐药是如何产生的，但治疗前原发性耐药的潜在机制仍不清楚。
进行单细胞谱系追踪和RNA测序，以鉴定HCC中对索拉非尼原发性耐药的谱系。采用差异基因表达分析来鉴定耐药谱系细胞的生物标志物。采用细胞活力和集落形成试验来评估S100A14对索拉非尼耐药的影响。进行免疫共沉淀（CO-IP）和质谱分析，以揭示S100A14在索拉非尼原发性耐药中的下游靶点和调控机制。进行体内小鼠异种移植实验，以评估S100A14或其相互作用蛋白谷氨酰胺酶（GLS）对HCC中索拉非尼原发性耐药的影响。
单细胞谱系追踪鉴定出一群对索拉非尼原发性耐药的细胞，并且确定一种钙结合蛋白S100A14是对索拉非尼原发性耐药的关键生物标志物。敲低S100A14可显著提高HCC细胞对索拉非尼治疗的敏感性。机制上，S100A14与GLS结合并阻断其Y308和S314位点的磷酸化，进而抑制其泛素化及随后的降解。通过稳定GLS，S100A14降低HCC细胞中的氧化应激，从而拮抗索拉非尼诱导的细胞凋亡。抑制S100A14或GLS可显著提高索拉非尼在体内对异种移植肿瘤的疗效。
我们的结果表明，S100A14通过稳定GLS以降低氧化应激，在促进对索拉非尼的原发性耐药中起关键作用，并作为一个潜在的治疗靶点，以提高索拉非尼在HCC患者中的疗效。
