Department of Neuromuscular Diseases, University College London Queen Square Institute of Neurology, Queen Square, London WC1N 3BG, UK.
The Francis Crick Institute, London NW1 1AT, UK.
Int J Mol Sci. 2021 Mar 23;22(6):3256. doi: 10.3390/ijms22063256.
Several studies have shown that human induced pluripotent stem cell (iPSC)-derivatives are essentially fetal in terms of their maturational status. Inducing ageing in iPSC-motor neuron (MN) models of amyotrophic lateral sclerosis (ALS) has the potential to capture pathology with higher fidelity and consequently improve translational success. We show here that the telomerase inhibitor BIBR1532, hypothesised to recapitulate the telomere attrition hallmark of ageing in iPSC-MNs, was in fact cytotoxic to feeder-free iPSCs when used at doses previously shown to be effective in iPSCs grown on a layer of mouse embryonic fibroblasts. Toxicity in feeder-free cultures was not rescued by co-treatment with Rho Kinase (ROCK) inhibitor (Y-27632). Moreover, the highest concentration of BIBR1532 compatible with continued iPSC culture proved insufficient to induce detectable telomerase inhibition. Our data suggest that direct toxicity by BIBR1532 is the most likely cause of iPSC death observed, and that culture methods may influence enhanced toxicity. Therefore, recapitulation of ageing hallmarks in iPSC-MNs, which might reveal novel and relevant human disease targets in ALS, is not achievable in feeder-free culture through the use of this small molecule telomerase inhibitor.
多项研究表明，从成熟状态来看，人类诱导多能干细胞（iPSC）衍生物本质上是胎儿。在肌萎缩侧索硬化症（ALS）的 iPSC-运动神经元（MN）模型中诱导衰老有可能更真实地捕捉到病理学特征，从而提高转化的成功率。我们在这里表明，端粒酶抑制剂 BIBR1532 假设可以再现 iPSC-MN 衰老的端粒损耗特征，当以前在一层小鼠胚胎成纤维细胞上生长的 iPSC 中证明有效的剂量使用时，实际上对无饲养层的 iPSC 具有细胞毒性。在无饲养层培养物中，与 Rho 激酶（ROCK）抑制剂（Y-27632）共同处理并不能挽救毒性。此外，与持续 iPSC 培养兼容的 BIBR1532 的最高浓度不足以诱导可检测到的端粒酶抑制。我们的数据表明，BIBR1532 的直接毒性很可能是观察到的 iPSC 死亡的最可能原因，并且培养方法可能会影响增强的毒性。因此，通过使用这种小分子端粒酶抑制剂，在无饲养层培养物中无法实现 iPSC-MN 衰老特征的再现，这可能会揭示 ALS 中新颖且相关的人类疾病靶点。
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