PhD Course in Reproductive and Developmental Sciences, University of Trieste, Trieste, Italy.
Department of Life Sciences, University of Trieste, Trieste, Italy.
Clin Pharmacol Ther. 2020 Aug;108(2):358-367. doi: 10.1002/cpt.1837. Epub 2020 May 9.
Ataxia telangiectasia (AT) and Aicardi-Goutières syndrome (AGS) are inherited disorders of immunity with prevalent neurological phenotype. Available treatments are only partially effective, and the prognosis is poor. Induced pluripotent stem cells (iPSCs) are obtained by reprogramming patient somatic cells, preserving the donor individual genetic heritage and creating patient-specific disease models, useful to investigate pathogenesis and drug effects and to develop precision therapies. The aim is to investigate the cytotoxicity of a panel of immunomodulators using iPSCs of patients with AT or different forms of AGS (AGS1, AGS2, and AGS7). iPSCs were obtained by reprogramming AT and AGS patients' cells and, as a control, the BJ normal human fibroblast line, using Sendai virus. Cytotoxic effects of two drugs proposed to treat respectively AT and AGS (dexamethasone and mepacrine) were tested by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay after 72 hours' exposure. Data were obtained also for other immunomodulatory drugs (thioguanine, mercaptopurine, thalidomide, and lenalidomide). Relative expression of genes involved in the tested drug pathways was analyzed. AGS7-derived iPSCs displayed altered viability when treated with a low dose of mepacrine and higher expression of cyclic guanosine monophosphate-adenosine monophosphate synthase, which is the main target for mepacrine action. AGS7-derived iPSCs were also more sensitive to thioguanine, while AGS2 and AT iPSCs were less sensitive to this medication than the BJ-iPSC. All iPSCs were equally sensitive to mercaptopurine and resistant to dexamethasone, thalidomide, and lenalidomide. This work establishes an innovative in vitro model that is useful to investigate the mechanisms of drugs potentially effective in AT and AGS.
共济失调毛细血管扩张症(AT)和 Aicardi-Goutières 综合征(AGS)是具有普遍神经表型的免疫遗传疾病。现有的治疗方法仅部分有效,预后较差。诱导多能干细胞(iPSC)通过重编程患者体细胞获得,保留供体个体的遗传遗产,并创建患者特异性疾病模型,可用于研究发病机制和药物作用,并开发精准治疗方法。目的是使用 AT 或不同形式的 AGS(AGS1、AGS2 和 AGS7)患者的 iPSC 研究一组免疫调节剂的细胞毒性。使用仙台病毒从 AT 和 AGS 患者的细胞中获得 iPSC,并作为对照从 BJ 正常人成纤维细胞系中获得 iPSC。用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐测定法在 72 小时暴露后测试两种拟用于治疗 AT 和 AGS 的药物(地塞米松和米帕林)的细胞毒性作用。还获得了其他免疫调节剂药物(硫鸟嘌呤、巯基嘌呤、沙利度胺和来那度胺)的数据。分析了参与测试药物途径的基因的相对表达。用低剂量米帕林处理 AGS7 衍生的 iPSC 时,其存活率发生改变,并且环鸟苷酸-腺苷酸合酶的表达增加,这是米帕林作用的主要靶标。AGS7 衍生的 iPSC 对硫鸟嘌呤也更敏感,而 AGS2 和 AT iPSC 比 BJ-iPSC 对这种药物的敏感性更低。所有 iPSC 对巯基嘌呤的敏感性相同,对地塞米松、沙利度胺和来那度胺均有耐药性。这项工作建立了一种创新的体外模型,可用于研究在 AT 和 AGS 中有效的潜在药物的作用机制。