Alternative oxidase encoded by sequence-optimized and chemically-modified RNA transfected into mammalian cells is catalytically active.
Alternative oxidase encoded by sequence-optimized and chemically-modified RNA transfected into mammalian cells is catalytically active.
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DOI：
10.1038/s41434-021-00235-z
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发表时间：
2022-12
期刊：
影响因子：
5.1
通讯作者：
Szibor, Marten
中科院分区：
文献类型：
--
作者：
Giordano, Luca;Aneja, Manish K.;Sommer, Natascha;Alebrahimdehkordi, Nasim;Seraji, Alireza;Weissmann, Norbert;Rudolph, Carsten;Plank, Christian;Jacobs, Howard T.;Szibor, Marten
植物和其他生物，但不是昆虫或脊椎动物，表达辅助呼吸酶交替氧化酶（AOX），当受损时绕过线粒体呼吸复合物III和/或IV。在哺乳动物模型中，来自玻璃海鞘的AOX的持续表达先前已被证明在缓解由呼吸链抑制产生的一些代谢应激同时加剧其他代谢应激方面是有效的。这意味着慢性AOX表达可能会改变或破坏协调适应性重塑所必需的代谢信号传导过程，表明其潜在的治疗用途可能仅限于急性病理，其中单疗程治疗就足够了。用于瞬时施用AOX的一种可能途径是编码AOX的核酸构建体。在这里，我们证明了AOX编码的化学修饰的RNA（cmRNA），序列优化的表达在哺乳动物细胞中，能够支持AOX表达在永生化小鼠胚胎成纤维细胞（iMEFs），人肺癌细胞（A549）和原代小鼠肺动脉平滑肌细胞（PASMCs）。AOX蛋白早在转染后3小时就可检测到，具有约4天的半衰期，并且具有催化活性，从而支持呼吸并防止呼吸抑制。我们的数据表明，AOX编码的cmRNA优化用于哺乳动物细胞代表了一个可行的路线，调查和可能治疗线粒体呼吸系统疾病。
Plants and other organisms, but not insects or vertebrates, express the auxiliary respiratory enzyme alternative oxidase (AOX) that bypasses mitochondrial respiratory complexes III and/or IV when impaired. Persistent expression of AOX from Ciona intestinalis in mammalian models has previously been shown to be effective in alleviating some metabolic stresses produced by respiratory chain inhibition while exacerbating others. This implies that chronic AOX expression may modify or disrupt metabolic signaling processes necessary to orchestrate adaptive remodeling, suggesting that its potential therapeutic use may be confined to acute pathologies, where a single course of treatment would suffice. One possible route for administering AOX transiently is AOX-encoding nucleic acid constructs. Here we demonstrate that AOX-encoding chemically-modified RNA (cmRNA), sequence-optimized for expression in mammalian cells, was able to support AOX expression in immortalized mouse embryonic fibroblasts (iMEFs), human lung carcinoma cells (A549) and primary mouse pulmonary arterial smooth muscle cells (PASMCs). AOX protein was detectable as early as 3 h after transfection, had a half-life of ~4 days and was catalytically active, thus supporting respiration and protecting against respiratory inhibition. Our data demonstrate that AOX-encoding cmRNA optimized for use in mammalian cells represents a viable route to investigate and possibly treat mitochondrial respiratory disorders.
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影响因子：
3.5
作者：
Humphrey, Dickon M.;Parsons, Richard B.;Hirth, Frank
通讯作者：
Hirth, Frank
DOI：
10.1534/g3.114.013946
发表时间：
2014-08-21
期刊：
G3 (Bethesda, Md.)
影响因子：
--
作者：
Kemppainen KK;Kemppainen E;Jacobs HT
通讯作者：
Jacobs HT
DOI：
10.1073/pnas.0810518105
发表时间：
2008-12-02
影响因子：
11.1
作者：
Perales-Clemente, Ester;Pilar Bayona-Bafaluy, Maria;Antonio Enriquez, Jose
通讯作者：
Antonio Enriquez, Jose
影响因子：
3.7
作者：
Lochmüller, H;Johns, T;Shoubridge, EA
通讯作者：
Shoubridge, EA
影响因子：
5.3
作者：
Andjelković A;Mordas A;Bruinsma L;Ketola A;Cannino G;Giordano L;Dhandapani PK;Szibor M;Dufour E;Jacobs HT
通讯作者：
Jacobs HT