Tsakiridis Evangelia E, Ahmadi Elham, Gautam Jaya, Hannah She Yi Ran, Fayyazi Russta, Lally James S V, Wang Simon, Di Pastena Fiorella, Valvano Celina M, Del Rosso Daniel, Biziotis Olga-Demetra, Meyers Brandon, Muti Paola, Tsakiridis Theodoros, Steinberg Gregory R
Centre for Metabolism, Obesity and Diabetes Research, McMaster University, 1280 Main Street West, Hamilton, ONT, L8S 4K1, Canada.
Division of Endocrinology and Metabolism, Department of Medicine, McMaster University, 1280 Main Street West, Hamilton, ONT, L8S 4K1, Canada.
JHEP Rep. 2025 Feb 14;7(5):101354. doi: 10.1016/j.jhepr.2025.101354. eCollection 2025 May.
BACKGROUND & AIMS: Metabolic dysfunction-associated steatohepatitis (MASH) is a growing cause of hepatocellular carcinoma (HCC) worldwide. The complex microenvironment of these tumors, characterized by metabolic dysfunction, hypoxia, steatosis, and fibrosis, limits the effectiveness of standard-of-care therapies, such as the multi-tyrosine kinase inhibitor lenvatinib (LEN). Salsalate (SAL), is a rheumatoid arthritis therapy that enhances fatty acid oxidation and reduces lipogenesis, fibrosis and cell proliferation pathways. We hypothesize that addition of SAL could improve the efficacy of LEN in MASH-HCC.
We assessed the efficacy of combination therapy using clinically relevant concentrations of LEN and SAL in human HCC cell models, orthotopic xenograft and MASH-HCC mouse models. In addition, assays assessing fatty acid oxidation and lipogenesis, protein immunoblotting and RNA-sequencing were used to understand mechanisms involved.
LEN + SAL synergistically suppressed the proliferation and clonogenic survival of cells ( ≤0.0001), prolonged survival in an orthotopic xenograft model ( = 0.02), and reduced angiogenesis, fibrosis, and steatosis ( ≤0.05) in a MASH-HCC model. These effects were associated with activation of AMPK and inhibition of the mTOR-HIF1α and Erk1/2 signaling pathways. RNA-sequencing analysis in both Hep3B cells and livers of the MASH-HCC mouse model revealed that SAL enhanced fatty acid oxidation and suppressed fibrosis and cell cycle progression, while LEN reduced angiogenesis with regulatory network analysis, suggesting a potential role for activating transcription factor 3 (ATF3) and ETS-proto-oncogene-1 (ETS-1).
These data indicate that combining LEN and SAL, which exert distinct effects leading to improvements in the liver microenvironment (steatosis, angiogenesis, and fibrosis) and inhibition of tumor proliferation, may have therapeutic potential for MASH-driven HCC.
Although rates of MASH-HCC are on the rise globally, standard-of-care multi-tyrosine kinase inhibitors and immunotherapy have limited efficacy in this HCC etiology. Metabolic targeting with SAL inhibits cancer growth kinetics while also alleviating drivers of MASH by increasing fatty acid oxidation and reducing lipogenesis and fibrosis. Combined LEN and SAL improved survival and MASH-HCC pathology in mouse models without adverse effects. Given that SAL is a safe, economical, and approved medication, this concept holds great translational potential that could provide a new treatment avenue for patients with unresected MASH-HCC.
代谢功能障碍相关脂肪性肝炎（MASH）是全球肝细胞癌（HCC）日益常见的病因。这些肿瘤的复杂微环境，其特征为代谢功能障碍、缺氧、脂肪变性和纤维化，限制了诸如多酪氨酸激酶抑制剂仑伐替尼（LEN）等标准治疗方法的疗效。双水杨酸酯（SAL）是一种用于治疗类风湿性关节炎的药物，可增强脂肪酸氧化并减少脂肪生成、纤维化和细胞增殖途径。我们假设添加SAL可提高LEN治疗MASH-HCC的疗效。
我们在人HCC细胞模型、原位异种移植模型和MASH-HCC小鼠模型中，使用临床相关浓度的LEN和SAL评估联合治疗的疗效。此外，还使用了评估脂肪酸氧化和脂肪生成的试验、蛋白质免疫印迹和RNA测序来了解其中涉及的机制。
LEN + SAL协同抑制细胞增殖和克隆形成存活（≤0.0001），延长原位异种移植模型中的生存期（P = 0.02），并减少MASH-HCC模型中的血管生成、纤维化和脂肪变性（≤0.05）。这些作用与AMPK激活以及mTOR-HIF1α和Erk1/2信号通路的抑制有关。对Hep3B细胞和MASH-HCC小鼠模型肝脏的RNA测序分析表明，SAL增强脂肪酸氧化并抑制纤维化和细胞周期进程，而LEN通过调控网络分析减少血管生成，提示激活转录因子3（ATF3）和ETS原癌基因-1（ETS-1）可能发挥作用。
这些数据表明，联合使用LEN和SAL可产生不同的作用，改善肝脏微环境（脂肪变性、血管生成和纤维化）并抑制肿瘤增殖，可能对MASH相关的HCC具有治疗潜力。
尽管全球MASH-HCC的发病率在上升，但标准治疗的多酪氨酸激酶抑制剂和免疫疗法在这种HCC病因中的疗效有限。用SAL进行代谢靶向可抑制癌症生长动力学，同时通过增加脂肪酸氧化和减少脂肪生成及纤维化来减轻MASH的驱动因素。联合使用LEN和SAL可改善小鼠模型的生存期和MASH-HCC病理状况，且无不良反应。鉴于SAL是一种安全、经济且已获批的药物，这一概念具有巨大的转化潜力，可为未切除的MASH-HCC患者提供新的治疗途径。
