Li Yiying, Zhang Junting, Sun Hanliang, Yu Xiufeng, Chen Yujie, Ma Cui, Zheng Xiaodong, Zhang Lixin, Zhao Xijuan, Jiang Yuan, Xin Wei, Wang Shanshan, Hu Jiye, Wang Mingge, Zhu Daling
Biopharmaceutical Key Laboratory of Heilongjiang Province, College of Pharmacy, Harbin Medical University, Harbin 150081, Heilongjiang Province, P.R. China.
Central Laboratory of Harbin Medical University (Daqing), Xinyang Road, Gaoxin District, Daqing 163319, P.R. China.
Mol Ther Nucleic Acids. 2022 May 21;28:920-934. doi: 10.1016/j.omtn.2022.05.033. eCollection 2022 Jun 14.
Pyroptosis is involved in pulmonary hypertension (PH); however, whether this process is regulated by long non-coding RNAs (lncRNAs) is unclear. Some lncRNAs encode peptides; therefore, whether the regulation of pyroptosis in PH depends on lncRNAs themselves or their encoded peptides needs to be explored. We aimed to characterize the role of the peptide RPS4XL encoded by lnc-Rps4l and its regulatory mechanisms during pyroptosis in PH. Transgenic mice overexpression of lnc-Rps4l was established to rescue the inhibition of hypoxia-induced pyroptosis in pulmonary artery smooth muscle cells (PASMCs). An adeno-associated virus 9 construct with a mutation in the open reading frame of lnc-Rps4l was used to verify that it could inhibit hypoxia-induced PASMCs pyroptosis through its encoded peptide RPS4XL. Glutathione S-transferase (GST) pull-down assays revealed that RPS4XL bound to HSC70, and microscale thermophoresis (MST) was performed to determine the HSC70 domain that interacted with RPS4XL. Through glycosylation site mutation, we confirmed that RPS4XL inhibited hypoxia-induced PASMCs pyroptosis by regulating HSC70 glycosylation. Our results showed that RPS4XL inhibits pyroptosis in a PH mouse model and hypoxic PASMCs by regulating HSC70 glycosylation. These results further clarify the important mechanism of vascular remodeling in PH pathology.
细胞焦亡参与肺动脉高压（PH）的发生过程；然而，这一过程是否受长链非编码RNA（lncRNA）调控尚不清楚。一些lncRNA可编码肽段；因此，PH中细胞焦亡的调控是取决于lncRNA本身还是其编码的肽段，仍有待探索。我们旨在明确lnc-Rps4l编码的肽段RPS4XL在PH细胞焦亡过程中的作用及其调控机制。构建lnc-Rps4l过表达转基因小鼠，以挽救缺氧诱导的肺动脉平滑肌细胞（PASMCs）焦亡抑制。使用lnc-Rps4l开放阅读框发生突变的腺相关病毒9构建体，以验证其可通过编码的肽段RPS4XL抑制缺氧诱导的PASMCs焦亡。谷胱甘肽S-转移酶（GST）下拉实验显示RPS4XL与HSC70结合，并进行微量热泳（MST）以确定与RPS4XL相互作用的HSC70结构域。通过糖基化位点突变，我们证实RPS4XL通过调节HSC70糖基化抑制缺氧诱导的PASMCs焦亡。我们的结果表明，RPS4XL通过调节HSC70糖基化抑制PH小鼠模型和缺氧PASMCs中的细胞焦亡。这些结果进一步阐明了PH病理中血管重塑的重要机制。
