Eloxx Pharmaceuticals, Watertown, MA, 02472, USA.
J Mol Med (Berl). 2023 Apr;101(4):375-385. doi: 10.1007/s00109-023-02291-x. Epub 2023 Feb 20.
Familial adenomatous polyposis (FAP) is a precancerous, colorectal disease characterized by hundreds to thousands of adenomatous polyps caused by mutations in the tumor suppressor gene adenomatous polyposis coli (APC). Approximately 30% of these mutations are premature termination codons (PTC), resulting in the production of a truncated, dysfunctional APC protein. Consequently, the β-catenin degradation complex fails to form in the cytoplasm, leading to elevated nuclear levels of β-catenin and unregulated β-catenin/wnt-pathway signaling. We present in vitro and in vivo data demonstrating that the novel macrolide, ZKN-0013, promotes read through of premature stop codons, leading to functional restoration of full-length APC protein. Human colorectal carcinoma SW403 and SW1417 cells harboring PTC mutations in the APC gene showed reduced levels of nuclear β-catenin and c-myc upon treatment with ZKN-0013, indicating that the macrolide-mediated read through of premature stop codons produced bioactive APC protein and inhibited the β-catenin/wnt-pathway. In a mouse model of adenomatous polyposis coli, treatment of APC mice with ZKN-0013 caused a significant decrease in intestinal polyps, adenomas, and associated anemia, resulting in increased survival. Immunohistochemistry revealed decreased nuclear β-catenin staining in the epithelial cells of the polyps in ZKN-0013-treated APC mice, confirming the impact on the β-catenin/wnt-pathway. These results indicate that ZKN-0013 may have therapeutic potential for the treatment of FAP caused by nonsense mutations in the APC gene. KEY MESSAGES: • ZKN-0013 inhibited the growth of human colon carcinoma cells with APC nonsense mutations. • ZKN-0013 promoted read through of premature stop codons in the APC gene. • In APC mice, ZKN-0013 treatment reduced intestinal polyps and their progression to adenomas. • ZKN-0013 treatment in APC mice resulted in reduced anemia and increased survival.
家族性腺瘤性息肉病（FAP）是一种癌前的结直肠疾病，其特征是由肿瘤抑制基因腺瘤性结肠息肉病基因（APC）的突变引起的数百到数千个腺瘤性息肉。这些突变约有 30%是提前终止密码子（PTC），导致产生截短的、功能失调的 APC 蛋白。因此，β-连环蛋白降解复合物在细胞质中无法形成，导致核内β-连环蛋白水平升高和β-连环蛋白/Wnt 信号通路的不受调节的信号转导。我们提供了体外和体内数据，证明新型大环内酯类药物 ZKN-0013 可促进提前终止密码子的通读，从而导致全长 APC 蛋白的功能恢复。携带 APC 基因中 PTC 突变的人结直肠癌细胞 SW403 和 SW1417 在用 ZKN-0013 处理后，核内β-连环蛋白和 c-myc 的水平降低，表明大环内酯类药物介导的提前终止密码子的通读产生了有生物活性的 APC 蛋白，并抑制了β-连环蛋白/Wnt 信号通路。在 APC 基因的腺瘤性结肠息肉病的小鼠模型中，用 ZKN-0013 治疗 APC 小鼠可显著减少肠息肉、腺瘤和相关的贫血，从而提高生存率。免疫组织化学显示，ZKN-0013 治疗的 APC 小鼠的息肉上皮细胞中的核β-连环蛋白染色减少，证实了对β-连环蛋白/Wnt 信号通路的影响。这些结果表明，ZKN-0013 可能具有治疗 APC 基因中无意义突变引起的 FAP 的治疗潜力。主要信息： • ZKN-0013 抑制 APC 无意义突变的人结肠癌细胞的生长。 • ZKN-0013 促进 APC 基因中提前终止密码子的通读。 • 在 APC 小鼠中，ZKN-0013 治疗减少了肠道息肉及其向腺瘤的进展。 • APC 小鼠中 ZKN-0013 的治疗导致贫血减少和生存率提高。
