Department of Gastroenterology and Oncology, Institute of Biomedical Sciences, Tokushima University Graduate School, 3-18-15 Kuramoto-cho, Tokushima, 770-8503, Japan.
Department of Community Medicine for Gastroenterology and Oncology, Tokushima University Graduate School of Biomedical Sciences, 3-18-15 Kuramoto-cho, Tokushima, 770-8503, Japan.
J Gastroenterol. 2024 Jan;59(1):11-23. doi: 10.1007/s00535-023-02052-0. Epub 2023 Nov 22.
Although the serrated-neoplasia pathway reportedly accounts for 15-30% of colorectal cancer (CRC), no studies on chemoprevention of sessile serrated lesions (SSLs) have been reported. We searched for effective compounds comprehensively from a large series of compounds by employing Connectivity Map (CMAP) analysis of SSL-specific gene expression profiles coupled with in vitro screening using SSL patient-derived organoids (PDOs), and validated their efficacy using a xenograft mouse model of SSL.
We generated SSL-specific gene signatures based on DNA microarray data, and applied them to CMAP analysis with 1309 FDA-approved compounds to select candidate compounds. We evaluated their inhibitory effects on SSL-PDOs using a cell viability assay. SSL-PDOs were orthotopically transplanted into NOG mice for in vivo evaluation. The signal transduction pathway was evaluated by gene expression profile and protein expression analysis.
We identified 221 compounds by employing CMAP analysis of SSL-specific signatures, which should cancel the gene signatures, and narrowed them down to 17 compounds. Cell viability assay using SSL-PDOs identified lansoprazole as having the lowest IC50 value (47 µM) among 17 compounds. When SSL-PDO was orthotopically transplanted into murine intestinal tract, the tumor grew gradually. Administration of lansoprazole to mice inhibited the growth of SSL xenograft whereas the tumor in control mice treated with vehicle alone grew gradually over time. The Ki67 index in xenograft lesions from the lansoprazole group was significantly lower compared with the control group. Cell cycle analysis of SSL-PDOs treated with lansoprazole exhibited a significant increase in G1 phase cell population. Microarray and protein analysis revealed that lansoprazole downregulated Skp2 expression and upregulated p27 expression in SSL-PDOs.
Our data strongly suggest that lansoprazole is the most effective chemopreventive agent against SSL, and that lansoprazole induces G1 cell cycle arrest by downregulating Skp2 and upregulating p27 in SSL cells.
虽然锯齿状肿瘤途径据称占结直肠癌（CRC）的 15-30%，但尚无关于 无蒂锯齿状病变（SSL）化学预防的研究。我们通过连接图谱（CMAP）分析 SSL 特异性基因表达谱与体外筛选相结合，从大量化合物中全面寻找有效的化合物，并用 SSL 患者来源的类器官（PDO）进行筛选，并使用 SSL 的异种移植小鼠模型验证其疗效。
我们基于 DNA 微阵列数据生成 SSL 特异性基因特征，并将其应用于 CMAP 分析，共分析了 1309 种 FDA 批准的化合物，以选择候选化合物。我们使用细胞活力测定法评估它们对 SSL-PDO 的抑制作用。将 SSL-PDO 原位移植到 NOG 小鼠中进行体内评价。通过基因表达谱和蛋白质表达分析评估信号转导途径。
我们通过对 SSL 特异性特征的 CMAP 分析鉴定出 221 种化合物，这些化合物应能消除基因特征，并将其缩小到 17 种化合物。使用 SSL-PDO 的细胞活力测定法鉴定出兰索拉唑是 17 种化合物中 IC50 值最低（47µM）的化合物。当 SSL-PDO 原位移植到小鼠肠道中时，肿瘤逐渐生长。兰索拉唑给药可抑制 SSL 异种移植物的生长，而单独用载体治疗的对照组肿瘤随时间推移逐渐生长。兰索拉唑组异种移植物病变的 Ki67 指数明显低于对照组。用兰索拉唑处理的 SSL-PDO 的细胞周期分析显示 G1 期细胞群显著增加。SSL-PDO 的微阵列和蛋白质分析显示，兰索拉唑下调了 SSL-PDO 中的 Skp2 表达并上调了 p27 表达。
我们的数据强烈表明，兰索拉唑是治疗 SSL 最有效的化学预防剂，兰索拉唑通过下调 Skp2 和上调 SSL 细胞中的 p27 诱导 G1 期细胞周期停滞。
