Essential roles of exosome and circRNA_101093 on ferroptosis desensitization in lung adenocarcinoma.
Essential roles of exosome and circRNA_101093 on ferroptosis desensitization in lung adenocarcinoma.
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外泌体和circRNA_101093对肺腺癌铁死亡脱敏的重要作用
DOI：
10.1002/cac2.12275
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发表时间：
2022-04
影响因子：
16.2
通讯作者：
Wang, Jiayi
中科院分区：
文献类型：
--
作者：
Zhang, Xiao;Xu, Yunhua;Ma, Lifang;Yu, Keke;Niu, Yongjie;Xu, Xin;Shi, Yi;Guo, Susu;Xue, Xiangfei;Wang, Yikun;Qiu, Shiyu;Cui, Jiangtao;Wang, Hong;Tian, Xiaoting;Miao, Yayou;Meng, Fanyu;Qiao, Yongxia;Yu, Yongchun;Wang, Jiayi
关键词：
对铁死亡的抵抗力是一种由铁依赖性脂质过氧化物过度积累引起的受调节细胞死亡，最近与肺腺癌（LUAD）有关。细胞内抗氧化系统是防止铁死亡所必需的。本研究的目的是研究细胞外系统是否以及如何使 LUAD 细胞对铁死亡不敏感。本研究使用已建立的人肺成纤维细胞MRC-5、WI38和人LUAD H1650、PC9、H1975、H358、A549和H1299细胞系、LUAD的肿瘤和匹配的正常邻近组织以及来自健康个体和LUAD患者的血浆。采用免疫组织化学和免疫印迹分析蛋白质表达，采用定量逆转录PCR分析mRNA表达。测量细胞活力、细胞死亡和脂质活性氧的产生，以评估对铁死亡的反应。使用透射电子显微镜观察外泌体。使用点击化学标记和共焦显微镜检测花生四烯酸 (AA) 的定位。使用 RNA Pull-down、RNA 免疫沉淀和光活化核糖核苷增强交联和免疫沉淀方法检测 RNA 和蛋白质之间的相互作用。蛋白质组学分析用于研究RNA调节蛋白，代谢组学分析用于分析代谢物。使用细胞来源的异种移植物、患者来源的异种移植物、细胞植入肺内 LUAD 小鼠模型和 LUAD 患者的血浆/组织标本来验证分子机制。来自 LUAD 患者的血浆外泌体特异性降低脂质过氧化并使 LUAD 细胞对铁死亡不敏感。一个可能的解释是，外泌体 circRNA_101093 (cir93) 在 LUAD 中维持细胞内 cir93 的升高，以调节 AA，AA 是一种对铁死亡相关的质膜过氧化增加至关重要的多不饱和脂肪酸。从机制上讲，cir93 与脂肪酸结合蛋白 3 (FABP3) 相互作用并增加脂肪酸结合蛋白 3 (FABP3)，后者转运 AA 并促进其与牛磺酸的反应。因此，整体 AA 减少，而 N-花生四烯酰牛磺酸（NAT，AA 和牛磺酸的产物）被诱导。值得注意的是，NAT 在抑制 AA 掺入质膜中的作用也被揭示。在临床前体内模型中，减少外泌体可改善基于铁死亡的治疗。外泌体和 cir93 对于使 LUAD 细胞对铁死亡脱敏至关重要，阻断外泌体可能有助于未来的 LUAD 治疗。
Resistance to ferroptosis, a regulated cell death caused by iron‐dependent excessive accumulation of lipid peroxides, has recently been linked to lung adenocarcinoma (LUAD). Intracellular antioxidant systems are required for protection against ferroptosis. The purpose of the present study was to investigate whether and how extracellular system desensitizes LUAD cells to ferroptosis. Established human lung fibroblasts MRC‐5, WI38, and human LUAD H1650, PC9, H1975, H358, A549, and H1299 cell lines, tumor and matched normal adjacent tissues of LUAD, and plasma from healthy individuals and LUAD patients were used in this study. Immunohistochemistry and immunoblotting were used to analyze protein expression, and quantitative reverse transcription‐PCR was used to analyze mRNA expression. Cell viability, cell death, and the lipid reactive oxygen species generation were measured to evaluate the responses to ferroptosis. Exosomes were observed using transmission electron microscope. The localization of arachidonic acid (AA) was detected using click chemistry labeling followed by confocal microscopy. Interactions between RNAs and proteins were detected using RNA pull‐down, RNA immunoprecipitation and photoactivatable ribonucleoside‐enhanced crosslinking and immunoprecipitation methods. Proteomic analysis was used to investigate RNA‐regulated proteins, and metabolomic analysis was performed to analyze metabolites. Cell‐derived xenograft, patient‐derived xenograft, cell‐implanted intrapulmonary LUAD mouse models and plasma/tissue specimens from LUAD patients were used to validate the molecular mechanism. Plasma exosome from LUAD patients specifically reduced lipid peroxidation and desensitized LUAD cells to ferroptosis. A potential explanation is that exosomal circRNA_101093 (cir93) maintained an elevation in intracellular cir93 in LUAD to modulate AA, a poly‐unsaturated fatty acid critical for ferroptosis‐associated increased peroxidation in the plasma membrane. Mechanistically, cir93 interacted with and increased fatty acid‐binding protein 3 (FABP3), which transported AA and facilitated its reaction with taurine. Thus, global AA was reduced, whereas N‐arachidonoyl taurine (NAT, the product of AA and taurine) was induced. Notably, the role of NAT in suppressing AA incorporation into the plasma membrane was also revealed. In pre‐clinical in vivo models, reducing exosome improved ferroptosis‐based treatment. Exosome and cir93 are essential for desensitizing LUAD cells to ferroptosis, and blocking exosome may be helpful for future LUAD treatment.
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影响因子：
14.8
作者：
Kagan VE;Mao G;Qu F;Angeli JP;Doll S;Croix CS;Dar HH;Liu B;Tyurin VA;Ritov VB;Kapralov AA;Amoscato AA;Jiang J;Anthonymuthu T;Mohammadyani D;Yang Q;Proneth B;Klein-Seetharaman J;Watkins S;Bahar I;Greenberger J;Mallampalli RK;Stockwell BR;Tyurina YY;Conrad M;Bayır H
通讯作者：
Bayır H
影响因子：
9
作者：
Guo S;Chen Y;Yang Y;Zhang X;Ma L;Xue X;Qiao Y;Wang J
通讯作者：
Wang J
影响因子：
3.4
作者：
ANDERSTAM, B;VACA, C;HARMSRINGDAHL, M
通讯作者：
HARMSRINGDAHL, M
影响因子：
7.7
作者：
Liin, Sara I.;Larsson, Johan E.;Larson, H. Peter
通讯作者：
Larson, H. Peter
影响因子：
16.2
作者：
Liu, Haijing;Zhang, Bo;Sun, Zhifu
通讯作者：
Sun, Zhifu