CRISPR/CasRx-Mediated RNA Knockdown Reveals That ACE2 Is Involved in the Regulation of Oligodendroglial Cell Morphological Differentiation.
CRISPR/CasRx-Mediated RNA Knockdown Reveals That ACE2 Is Involved in the Regulation of Oligodendroglial Cell Morphological Differentiation.
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CRISPR/CasRx介导的RNA敲低揭示ACE 2参与调节少突胶质细胞形态分化
DOI：
10.3390/ncrna8030042
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发表时间：
2022-06-06
期刊：
影响因子：
4.3
通讯作者：
Yamauchi, Junji
中科院分区：
文献类型：
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作者：
Kato, Yukino;Tago, Kenji;Fukatsu, Shoya;Okabe, Miyu;Shirai, Remina;Oizumi, Hiroaki;Ohbuchi, Katsuya;Yamamoto, Masahiro;Mizoguchi, Kazushige;Miyamoto, Yuki;Yamauchi, Junji
血管紧张素转换酶2（ACE 2）在催化血管紧张素II转化为血管紧张素（1-7）中起作用，血管紧张素（1-7）通常会抵消肾素-血管紧张素系统。ACE 2不仅在外周组织如心脏和肾脏的细胞中表达，而且在中枢神经系统（CNS）的细胞中表达。此外，ACE 2作为严重急性呼吸综合征冠状病毒2（SARS-CoV-2）进入所需的受体，其结合导致ACE 2蛋白本身的内吞再循环和可能的降解。中枢神经系统中SARS-CoV-2的靶细胞之一是少突胶质细胞（少突胶质细胞），它们用称为髓磷脂膜的分化质膜包裹神经元轴突。在这里，我们第一次描述了ACE 2在FBD-102 b细胞中的作用，该细胞被用作少突胶质细胞的分化模型。出乎意料的是，用CasRx介导的gRNA或同源siRNA对ACE 2的RNA敲低促进了少突胶质细胞形态分化，其中分化和/或髓磷脂标志物蛋白的表达或磷酸化水平增加，表明ACE 2在形态分化中的负面作用。值得注意的是，ACE 2的细胞内区域优先与活性GTP结合形式的Ras相互作用。因此，在亲和沉淀试验中，ACE 2的敲低相对增加了GTP结合的Ras。事实上，Ras的抑制导致形态分化和标记蛋白的表达或磷酸化水平降低，证实了Ras在分化中的积极作用。这些结果表明ACE 2本身作为少突胶质细胞形态分化的负调节剂的作用，新添加ACE 2的少突胶质细胞形态发生的调节剂以及Ras结合蛋白的列表。这些发现可能有助于我们理解为什么SARS-CoV-2会在CNS中引起病理效应。
Angiotensin-converting enzyme 2 (ACE2) plays a role in catalyzing angiotensin II conversion to angiotensin (1–7), which often counteracts the renin-angiotensin system. ACE2 is expressed not only in the cells of peripheral tissues such as the heart and kidney, but also in those of the central nervous system (CNS). Additionally, ACE2 acts as the receptor required for the entry of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), whose binding leads to endocytotic recycling and possible degradation of the ACE2 proteins themselves. One of the target cells for SARS-CoV-2 in the CNS is oligodendrocytes (oligodendroglial cells), which wrap neuronal axons with their differentiated plasma membranes called myelin membranes. Here, for the first time, we describe the role of ACE2 in FBD-102b cells, which are used as the differentiation models of oligodendroglial cells. Unexpectedly, RNA knockdown of ACE2 with CasRx-mediated gRNA or the cognate siRNA promoted oligodendroglial cell morphological differentiation with increased expression or phosphorylation levels of differentiation and/or myelin marker proteins, suggesting the negative role of ACE2 in morphological differentiation. Notably, ACE2′s intracellular region preferentially interacted with the active GTP-bound form of Ras. Thus, knockdown of ACE2 relatively increased GTP-bound Ras in an affinity-precipitation assay. Indeed, inhibition of Ras resulted in decreasing both morphological differentiation and expression or phosphorylation levels of marker proteins, confirming the positive role of Ras in differentiation. These results indicate the role of ACE2 itself as a negative regulator of oligodendroglial cell morphological differentiation, newly adding ACE2 to the list of regulators of oligodendroglial morphogenesis as well as of Ras-binding proteins. These findings might help us to understand why SARS-CoV-2 causes pathological effects in the CNS.