Exploring the factors underlying remyelination arrest by studying the post‐transcriptional regulatory mechanisms of cystatin F gene
Exploring the factors underlying remyelination arrest by studying the post‐transcriptional regulatory mechanisms of cystatin F gene
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DOI：
10.1111/jnc.15190
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发表时间：
2020-09
影响因子：
4.7
通讯作者：
Jiayi Li;W. Durose;J. Ito;A. Kakita;Y. Iguchi;M. Katsuno;Kazuo Kunisawa;Takeshi Shimizu;K. Ikenaka
中科院分区：
文献类型：
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作者：
Jiayi Li;W. Durose;J. Ito;A. Kakita;Y. Iguchi;M. Katsuno;Kazuo Kunisawa;Takeshi Shimizu;K. Ikenaka
髓鞘再生在决定脱髓鞘疾病的命运方面起着重要作用。然而，它在慢性疾病状态下被抑制。胱抑素 F 是一种类似木瓜蛋白酶的溶酶体半胱氨酸蛋白酶抑制剂，是脱髓鞘和髓鞘再生的重要调节剂。使用半合子蛋白脂质蛋白转基因4e（PLP4e/‐）小鼠（一种慢性脱髓鞘动物模型），我们发现胱抑素F mRNA表达在2.5月龄时被诱导，并在脱髓鞘早期阶段上调，但在慢性阶段显着下降。接下来我们研究了胱抑素 F 调节因子作为慢性脱髓鞘疾病中髓鞘再生停滞的潜在机制。我们使用 CysF-STOP-tetO::Iba-mtTA 小鼠模型，其中胱抑素 F 基因表达由四环素操纵子驱动。有趣的是，我们发现强制胱抑素 F mRNA 过度表达最终会减少。我们的研究结果表明胱抑素 F 表达在转录后受到调节。接下来，我们将胚胎致死、视力异常、果蝇样 RNA 结合蛋白 1 (ELAVL-1) 和 miR29a 分别确定为胱抑素 F mRNA 稳定和不稳定因子。这些作用在体外 NIH3T3 细胞中得到证实。使用人类多发性硬化症患者的死后斑块样本，我们还证实 ELAVL-1 表达与先前报道的胱抑素 F 表达模式高度相关。这些数据表明 ELAVL-1 和 miR29a 在调节胱抑素 F 表达中的重要作用。此外，它们还为脱髓鞘疾病的潜在治疗靶点提供了新的见解。
Remyelination plays an important role in determining the fate of demyelinating disorders. However, it is arrested during chronic disease states. Cystatin F, a papain‐like lysosomal cysteine proteinase inhibitor, is a crucial regulator of demyelination and remyelination. Using hemizygous proteolipid protein transgenic 4e (PLP4e/‐) mice, an animal model of chronic demyelination, we found that cystatin F mRNA expression was induced at 2.5 months of age and up‐regulated in the early phase of demyelination, but significantly decreased in the chronic phase. We next investigated cystatin F regulatory factors as potential mechanisms of remyelination arrest in chronic demyelinating disorders. We used the CysF‐STOP‐tetO::Iba‐mtTA mouse model, in which cystatin F gene expression is driven by the tetracycline operator. Interestingly, we found that forced cystatin F mRNA over‐expression was eventually decreased. Our findings show that cystatin F expression is modulated post‐transcriptionally. We next identified embryonic lethal, abnormal vision, drosophila like RNA‐binding protein 1 (ELAVL‐1), and miR29a as cystatin F mRNA stabilizing and destabilizing factors, respectively. These roles were confirmed in vitro in NIH3T3 cells. Using postmortem plaque samples from human multiple sclerosis patients, we also confirmed that ELAVL‐1 expression was highly correlated with the previously reported expression pattern of cystatin F. These data indicate the important roles of ELAVL‐1 and miR29a in regulating cystatin F expression. Furthermore, they provide new insights into potential therapeutic targets for demyelinating disorders.