LincR-PPP2R5C通过PPP2R5C/PP2A在过敏性哮喘中形成RNA-DNA三链体促进Th2细胞分化。
LincR-PPP2R5C Promotes Th2 Cell Differentiation Through PPP2R5C/PP2A by Forming an RNA-DNA Triplex in Allergic Asthma.
作者信息
Ji Ningfei, Chen Zhongqi, Wang Zhengxia, Sun Wei, Yuan Qi, Zhang Xijie, Jia Xinyu, Wu Jingjing, Jiang Jingxian, Song Meijuan, Xu Tingting, Liu Yanan, Ma Qiyun, Sun Zhixiao, Bao Yanmin, Zhang Mingshun, Huang Mao
机构信息
Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
Department of Respiratory and Critical Care Medicine, Xishan People's Hospital of Wuxi City, Wuxi Branch of Zhongda Hospital Affiliate to Southeast University, Wuxi, China.
出版信息
Allergy Asthma Immunol Res. 2024 Jan;16(1):71-90. doi: 10.4168/aair.2024.16.1.71.
PURPOSE
The roles and mechanisms of long noncoding RNAs (lncRNAs) in T helper 2 (Th2) differentiation from allergic asthma are poorly understood. We aimed to explore a novel lncRNA, LincR-protein phosphatase 2 regulatory subunit B' gamma (PPP2R5C), in Th2 differentiation in a mouse model of asthma.
METHODS
LincR-PPP2R5C from RNA-seq data of CD4 T cells of asthma-like mice were validated and confirmed by quantitative reverse transcription polymerase chain reaction, northern blotting, nuclear and cytoplasmic separation, and fluorescence hybridization (FISH). Lentiviruses encoding LincR-PPP2R5C or shRNA were used to overexpress or silence LincR-PPP2R5C in CD4 T cells. The interactions between LincR-PPP2R5C and PPP2R5C were explored with western blotting, chromatin isolation by RNA purification assay, and fluorescence resonance energy transfer. An ovalbumin-induced acute asthma model in knockout (KO) mice (LincR-PPP2R5C KO, CD4 conditional LincR-PPP2R5C KO) was established to explore the roles of LincR-PPP2R5C in Th2 differentiation.
RESULTS
LncR-PPP2R5C was significantly higher in CD4 T cells from asthmatic mice and Th2 cells . The lentivirus encoding LincR-PPP2R5C suppressed Th1 differentiation; in contrast, the short hairpin RNA (shRNA) lentivirus decreased LincR-PPP2R5C and Th2 differentiation. Mechanistically, LincR-PPP2R5C deficiency suppressed the phosphatase activity of the protein phosphatase 2A (PP2A) holocomplex, resulting in a decline in Th2 differentiation. The formation of an RNA-DNA triplex between LincR-PPP2R5C and the PPP2R5C promoter enhanced PPP2R5C expression and activated PP2A. LincR-PPP2R5C KO and CD4 conditional KO decreased Th2 differentiation, airway hyperresponsiveness and inflammatory responses.
CONCLUSIONS
LincR-PPP2R5C regulated PPP2R5C expression and PP2A activity by forming an RNA-DNA triplex with the PPP2R5C promoter, leading to Th2 polarization in a mouse model of acute asthma. Our data presented the first definitive evidence of lncRNAs in the regulation of Th2 cells in asthma.
目的
长链非编码RNA（lncRNA）在过敏性哮喘辅助性T细胞2（Th2）分化中的作用和机制尚不清楚。我们旨在探讨一种新型lncRNA，即LincR-蛋白磷酸酶2调节亚基Bγ（PPP2R5C）在哮喘小鼠模型Th2分化中的作用。
方法
通过定量逆转录聚合酶链反应、Northern印迹、核质分离和荧光杂交（FISH）对哮喘样小鼠CD4 T细胞RNA测序数据中的LincR-PPP2R5C进行验证和确认。使用编码LincR-PPP2R5C或短发夹RNA（shRNA）的慢病毒在CD4 T细胞中过表达或沉默LincR-PPP2R5C。通过蛋白质印迹、RNA纯化法染色质分离和荧光共振能量转移探索LincR-PPP2R5C与PPP2R5C之间的相互作用。建立卵清蛋白诱导的基因敲除（KO）小鼠（LincR-PPP2R5C KO、CD4条件性LincR-PPP2R5C KO）急性哮喘模型，以探讨LincR-PPP2R5C在Th2分化中的作用。
结果
哮喘小鼠和Th2细胞的CD4 T细胞中LncR-PPP2R5C显著升高。编码LincR-PPP2R5C的慢病毒抑制Th1分化；相反，短发夹RNA（shRNA）慢病毒降低LincR-PPP2R5C和Th2分化。机制上，LincR-PPP2R5C缺陷抑制了蛋白磷酸酶2A（PP2A）全酶复合物的磷酸酶活性，导致Th2分化下降。LincR-PPP2R5C与PPP2R5C启动子之间形成RNA-DNA三链体增强了PPP2R5C表达并激活了PP2A。LincR-PPP2R5C KO和CD4条件性KO降低了Th2分化、气道高反应性和炎症反应。
结论
LincR-PPP2R5C通过与PPP2R5C启动子形成RNA-DNA三链体调节PPP2R5C表达和PP2A活性，导致急性哮喘小鼠模型中的Th2极化。我们的数据首次明确证明了lncRNA在哮喘中对Th2细胞的调节作用。
Zotero 插件