Tumor metastasis is a complex process. A better understanding of the mechanistic underpinnings of the metastatic cascade could uncover approaches to better prevent and target metastases. Here, we integrated transcriptomic profiling in primary esophageal squamous cell carcinoma (ESCC) and matched lymph node metastatic tissues and genome-wide CRISPR/Cas9 functional screening to identify the key drivers of cancer metastasis. Anoctamin 1 (ANO1) expression was found to be elevated in primary tumors and further increased in metastatic tissues. High expression of ANO1 was associated with poor patient survival, and ANO1 overexpression promoted cancer invasion and metastasis. Mechanistically, ANO1 inactivated the LXR pathway and increased intracellular cholesterol accumulation by interacting with JUN to repress the transcription of cholesterol hydroxylase, CYP27A1. In a paracrine manner, IL1β secreted from ANO1-expressing ESCC cancer cells activated NF-κB signaling in fibroblasts to stimulate production of the chemokine CCL1, which in turn enhanced invasion of ESCC cancer cells. Furthermore, molecular docking and in silico screening of 1,500,000 small molecules identified K786–4469 as a lead compound that targeted ANO1 and suppressed cancer metastasis in vitro and in vivo. Overall, these findings uncover a noncanonical function of ANO1 in regulating cholesterol metabolism and fibroblast function, revealing ANO1 as a potential prognostic biomarker and therapeutic target for metastatic ESCC. Significance: Metastatic cancer cells upregulate ANO1 to activate cell-intrinsic and -extrinsic mechanisms that alter cholesterol metabolism and stimulate fibroblasts, which can be targeted with ANO1 inhibitors to inhibit metastatic growth. See related commentary by Singh and Mehla, p. 1759

中文翻译：

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ANO1 重新编程胆固醇代谢和肿瘤微环境以促进癌症转移

肿瘤转移是一个复杂的过程。更好地了解转移级联的机制基础可以发现更好地预防和靶向转移的方法。在这里，我们整合了原发性食管鳞状细胞癌 (ESCC) 和匹配的淋巴结转移组织的转录组分析以及全基因组 CRISPR/Cas9 功能筛选，以确定癌症转移的关键驱动因素。Anoctamin 1 (ANO1) 的表达在原发性肿瘤中升高，在转移组织中进一步升高。ANO1的高表达与患者较差的生存率相关，ANO1的过度表达会促进癌症的侵袭和转移。从机制上讲，ANO1 通过与 JUN 相互作用抑制胆固醇羟化酶的转录，使 LXR 通路失活并增加细胞内胆固醇的积累，CYP27A1。以旁分泌方式，表达 ANO1 的 ESCC 癌细胞分泌的 IL1β 激活成纤维细胞中的 NF-κB 信号传导，刺激趋化因子 CCL1 的产生，从而增强 ESCC 癌细胞的侵袭。此外，对 1,500,000 个小分子进行分子对接和计算机筛选，确定 K786-4469 是一种先导化合物，可靶向 ANO1 并在体外和体内抑制癌症转移。总的来说，这些发现揭示了 ANO1 在调节胆固醇代谢和成纤维细胞功能中的非典型功能，揭示了 ANO1 作为转移性 ESCC 的潜在预后生物标志物和治疗靶点。意义：转移性癌细胞上调 ANO1，激活细胞内在和外在机制，改变胆固醇代谢并刺激成纤维细胞，ANO1 抑制剂可靶向抑制转移生长。参见 Singh 和 Mehla 的相关评论，第 17 页。第1759章