Inhibition of ALK-Signaling Overcomes STRN-ALK-Induced Downregulation of the Sodium Iodine Symporter and Restores Radioiodine Uptake in Thyroid Cells.
Inhibition of ALK-Signaling Overcomes STRN-ALK-Induced Downregulation of the Sodium Iodine Symporter and Restores Radioiodine Uptake in Thyroid Cells.
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抑制 ALK 信号传导可克服 STRN-ALK 诱导的钠碘同向转运蛋白下调，并恢复甲状腺细胞对放射性碘的摄取。
DOI：
10.1089/thy.2022.0533
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Nikiforov,YuriE
中科院分区：
文献类型：
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作者：
Nikitski,AlyaksandrV;Condello,Vincenzo;Divakaran,SaurabhS;Nikiforov,YuriE
背景：放射性碘（RAI）通常用于甲状腺癌治疗，尽管其治疗益处仅限于嗜碘肿瘤。RAI难治性疾病随着肿瘤去分化而发展，涉及钠-碘同向转运体（NIS）的丢失。由ALK融合驱动的甲状腺癌倾向于去分化，靶向ALK抑制是否可以增强这些肿瘤中的RAI摄取仍然未知。本研究的目的是确定ALK融合驱动的甲状腺癌进展过程中NIS的表达水平，评估ALK激活对NIS介导的RAI摄取的影响，并测试其modulation.Methods药理学选项：使用STRN-ALK驱动的甲状腺癌小鼠模型分析了NIS在ALK驱动的癌变不同阶段的表达。对于体外实验，使用PCCL 3正常甲状腺细胞产生了强力霉素诱导的STRN-ALK表达系统。通过定量逆转录聚合酶链反应、蛋白质印迹、免疫荧光、RNA测序和基因集途径分析评价了STRN-ALK诱导的效应。用131 I测量RAI摄取。治疗实验用FDA批准的ALK抑制剂（crizotinib和ceritinib）、MEK抑制剂selumetinib和JAK 1/2抑制剂ruxolitinib.Results：我们发现Nis下调发生在ALK驱动的甲状腺癌发生的早期，甚至在高分化癌阶段，在低分化甲状腺癌中完全丧失。甲状腺细胞中的急性STRN-ALK表达导致MAPK、JAK/STAT 3和PI 3 K/AKT/mTOR信号输出增加，与大多数甲状腺分化和碘代谢/转运基因（包括Slc 5a 5（Nis）、Foxe 1、Dio 1、Duox 1/2、Duoxa 2、Glis 3、Slc 5a 8和Tg）的ALK依赖性显著下调相关。此外，甲状腺细胞中的STRN-ALK表达诱导了膜NIS的显著损失和NIS介导的RAI摄取的四倍降低，这被ALK抑制剂克唑替尼和色瑞替尼逆转。此外，一个强大的剂量依赖性恢复NIS与其膜再分布在STRN-ALK表达的甲状腺细胞后，观察到抑制MAPK信号与司美替尼，表现出与JAK 1/2抑制剂ruxolitinib.Conclusions的累积效应：本临床前研究的结果表明，ALK融合诱导的NIS下调，RAI难治性的先决条件，可以在甲状腺细胞逆转，无论是直接抑制ALK或其下游信号通路。
Background:Radioiodine (RAI) is commonly used for thyroid cancer treatment, although its therapeutic benefits are restricted to iodine-avid tumors. The RAI-refractory disease develops with tumor dedifferentiation involving loss of sodium-iodine symporter (NIS). Thyroid cancers driven by ALK fusions are prone to dedifferentiation, and whether targeted ALK inhibition may enhance RAI uptake in these tumors remains unknown. The aim of this study was to determine the levels of NIS expression during the progression of ALK fusion-driven thyroid cancer, assess the effects of ALK activation on NIS-mediated RAI uptake, and test pharmacological options for its modulation.Methods:The expression of NIS at different stages of ALK-driven carcinogenesis was analyzed using a mouse model of STRN-ALK-driven thyroid cancer. Forin vitroexperiments, a system of doxycycline-inducible expression of STRN-ALK was generated using PCCL3 normal thyroid cells. The STRN-ALK-induced effects were evaluated with quantitative reverse transcription polymerase chain reaction, Western blot, immunofluorescence, RNA sequencing, and gene sets pathways analyses. RAI uptake was measured using131I. Treatment experiments were done with FDA-approved ALK inhibitors (crizotinib and ceritinib), MEK inhibitor selumetinib, and JAK1/2 inhibitor ruxolitinib.Results:We found thatNisdownregulation occurred early in ALK-driven thyroid carcinogenesis, even at the stage of well-differentiated cancer, with a complete loss in poorly differentiated thyroid carcinomas. Acute STRN-ALK expression in thyroid cells resulted in increased MAPK, JAK/STAT3, and PI3K/AKT/mTOR signaling outputs associated with significant ALK-dependent downregulation of the majority of thyroid differentiation and iodine metabolism/transport genes, includingSlc5a5(Nis),Foxe1,Dio1,Duox1/2,Duoxa2,Glis3,Slc5a8, andTg. Moreover, STRN-ALK expression in thyroid cells induced a significant loss of membranous NIS and a fourfold decrease of the NIS-mediated RAI uptake, which were reversed by ALK inhibitors crizotinib and ceritinib. In addition, a strong dose-dependent restoration of NIS with its membranous redistribution in STRN-ALK-expressing thyroid cells was observed after inhibition of MAPK signaling with selumetinib, which exhibited a cumulative effect with JAK1/2 inhibitor ruxolitinib.Conclusions:The findings of this preclinical study showed that ALK fusion-induced downregulation of NIS, the prerequisite of RAI refractoriness, could be reversed in thyroid cells by either direct inhibition of ALK or its downstream signaling pathways.