A genetically encoded fluorescent biosensor for detecting itaconate with subcellular resolution in living macrophages.
A genetically encoded fluorescent biosensor for detecting itaconate with subcellular resolution in living macrophages.
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DOI：
10.1038/s41467-022-34306-5
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发表时间：
2022-11-04
影响因子：
16.6
通讯作者：
Li, Xinjian
中科院分区：
文献类型：
--
作者：
Sun, Pengkai;Zhang, Zhenxing;Wang, Bin;Liu, Caiyun;Chen, Chao;Liu, Ping;Li, Xinjian
衣康酸是一种新发现的内源性代谢物，在先天免疫反应中促进抗炎程序，但由于可用的衣康酸监测技术的局限性，其作用的确切机制仍然知之甚少。在这里，我们开发并验证了一种遗传编码的衣康酸荧光生物传感器BioITA，用于直接监测活巨噬细胞亚细胞区室中的衣康酸动态。利用BioITA，我们监测衣康酸在脂多糖（LPS）刺激下调节衣康酸运输和代谢的动态。此外，我们发现STING激活诱导衣康酸产生，并且将表达胞质BioITA的aav注射到小鼠体内，可以直接报道在lps注射小鼠的活化巨噬细胞中衣康酸水平升高。因此，BioITA能够在活的巨噬细胞中实现衣康酸的亚细胞分辨率成像。衣康酸已被确定为活化巨噬细胞产生的免疫调节代谢物，但缺乏在活细胞中检测衣康酸的方法。在这里，作者开发了一种名为BioITA的荧光生物传感器，用于检测活巨噬细胞亚细胞区室中的衣康酸。
Itaconate is a newly discovered endogenous metabolite promoting an anti-inflammatory program during innate immune response, but the precise mechanisms underlying its effect remains poorly understood owing primarily to the limitations of available itaconate-monitoring techniques. Here, we develop and validate a genetically encoded fluorescent itaconate biosensor, BioITA, for directly monitoring itaconate dynamics in subcellular compartments of living macrophages. Utilizing BioITA, we monitor the itaconate dynamics in response to lipopolysaccharide (LPS) stimulation in the context of modulating itaconate transportation and metabolism. Moreover, we show that STING activation induces itaconate production, and injection of AAVs expressing cytosolic BioITA into mice allows directly reporting elevation of itaconate level in activated macrophages derived from LPS-injected mice. Thus, BioITA enables subcellular resolution imaging of itaconate in living macrophages. Itaconate has been identified as an immunomodulatory metabolite produced by activated macrophages, but methods for detecting itaconate in live cells are lacking. Here, the authors develop a fluorescent biosensor named BioITA for detecting itaconate in subcellular compartments of living macrophages.
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影响因子：
29
作者：
Lampropoulou V;Sergushichev A;Bambouskova M;Nair S;Vincent EE;Loginicheva E;Cervantes-Barragan L;Ma X;Huang SC;Griss T;Weinheimer CJ;Khader S;Randolph GJ;Pearce EJ;Jones RG;Diwan A;Diamond MS;Artyomov MN
通讯作者：
Artyomov MN
影响因子：
6.7
作者：
Naujoks J;Tabeling C;Dill BD;Hoffmann C;Brown AS;Kunze M;Kempa S;Peter A;Mollenkopf HJ;Dorhoi A;Kershaw O;Gruber AD;Sander LE;Witzenrath M;Herold S;Nerlich A;Hocke AC;van Driel I;Suttorp N;Bedoui S;Hilbi H;Trost M;Opitz B
通讯作者：
Opitz B
影响因子：
29
作者：
Runtsch, Marah C.;Angiari, Stefano;O'Neill, Luke A. J.
通讯作者：
O'Neill, Luke A. J.
影响因子：
29
作者：
Hooftman, Alexander;Angiari, Stefano;O'Neill, Luke A. J.
通讯作者：
O'Neill, Luke A. J.
影响因子：
15
作者：
Strelko, Cheryl L.;Lu, Wenyun;Dufort, Fay J.;Seyfried, Thomas N.;Chiles, Thomas C.;Rabinowitz, Joshua D.;Roberts, Mary F.
通讯作者：
Roberts, Mary F.