使用瑞戈非尼（一种多激酶抑制剂）治疗转移性结直肠癌（mCRC）显示总体生存率略有改善，但与严重的毒性相关。因此，为了减少瑞戈非尼引起的毒性，我们将低浓度的瑞戈非尼与双重 JAK/HDAC 小分子抑制剂 (JAK/HDACi) 一起使用，以利用 JAK 和 HDAC 抑制的优势来增强抗肿瘤活性。使用CRC模型评估联合治疗的疗效和安全性。使用具有不同遗传背景的正常结肠和CRC细胞测试JAK/HDACi、瑞戈非尼及其组合的细胞毒性。进行了 Kinomic、ATAC-seq、RNA-seq、细胞周期和细胞凋亡分析，以评估受组合影响的细胞功能/分子改变。使用患者来源的异种移植物（PDX）和结直肠癌的实验转移模型评估了该组合的疗效。为了评估肿瘤、其微环境和免疫反应调节之间的相互作用，使用了 MC38 同基因小鼠。联合治疗降低了细胞活力； JAK、STAT3、EGFR 和其他关键激酶的磷酸化；并抑制组蛋白 H3K9、H4K8 和 α 微管蛋白的脱乙酰化。它诱导细胞周期停滞在 G0-G1 期和 CRC 细胞凋亡。全转录组分析表明，联合治疗调节参与细胞凋亡、细胞外基质-受体相互作用和粘着斑途径的分子。它可以协同减少 PDX 肿瘤的生长和实验性转移，并且在同基因小鼠模型中，该治疗增强了抗肿瘤免疫反应，CD45 和细胞毒性细胞的浸润增加就证明了这一点。药代动力学研究表明，联合用药可增加瑞戈非尼的生物利用度。联合治疗比单独使用瑞戈非尼或JAK/HDACi更有效，并且毒性最小。有必要进行一项临床试验来评估这种组合治疗 mCRC 的效果。© 2024。作者。

Treatment with regorafenib, a multiple-kinase inhibitor, to manage metastatic colorectal cancers (mCRCs) shows a modest improvement in overall survival but is associated with severe toxicities. Thus, to reduce regorafenib-induced toxicity, we used regorafenib at low concentration along with a dual JAK/HDAC small-molecule inhibitor (JAK/HDACi) to leverage the advantages of both JAK and HDAC inhibition to enhance antitumor activity. The therapeutic efficacy and safety of the combination treatment was evaluated with CRC models.The cytotoxicity of JAK/HDACi, regorafenib, and their combination were tested with normal colonic and CRC cells exhibiting various genetic backgrounds. Kinomic, ATAC-seq, RNA-seq, cell cycle, and apoptosis analyses were performed to evaluate the cellular functions/molecular alterations affected by the combination. Efficacy of the combination was assessed using patient-derived xenograft (PDX) and experimental metastasis models of CRC. To evaluate the interplay between tumor, its microenvironment, and modulation of immune response, MC38 syngeneic mice were utilized.The combination therapy decreased cell viability; phosphorylation of JAKs, STAT3, EGFR, and other key kinases; and inhibited deacetylation of histone H3K9, H4K8, and alpha tubulin proteins. It induced cell cycle arrest at G0-G1 phase and apoptosis of CRC cells. Whole transcriptomic analysis showed that combination treatment modulated molecules involved in apoptosis, extracellular matrix-receptor interaction, and focal adhesion pathways. It synergistically reduces PDX tumor growth and experimental metastasis, and, in a syngeneic mouse model, the treatment enhances the antitumor immune response as evidenced by higher infiltration of CD45 and cytotoxic cells. Pharmacokinetic studies showed that combination increased the bioavailability of regorafenib.The combination treatment was more effective than with regorafenib or JAK/HDACi alone, and had minimal toxicity. A clinical trial to evaluate this combination for treatment of mCRCs is warranted.© 2024. The Author(s).