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Long non-coding RNA nuclear paraspeckle assembly transcript 1 promotes activation of T helper 2 cells via inhibiting STAT6 ubiquitination

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Abstract

T helper (Th) 2 cell-medicated immune response participates in various immune diseases, including systemic lupus erythematosus (SLE). Long non-coding RNA nuclear paraspeckle assembly transcript 1 (NEAT1) has been reported to be associated with T helper 2 (Th2) cell activation. Here, we demonstrated the molecular mechanism of NEAT1 in regulating Th2 cell activation. We found that NEAT1 was located in nucleus. NEAT1 overexpression promoted the levels of Th2-related cytokines IL-4, IL-5 and IL-13 in CD4+ T cells. Moreover, NEAT1 up-regulation reduced Th1-related cytokine INF-γ production and enhanced the levels of Th17-related cytokines IL-17 in CD4+ T cells. STAT6 deficiency reduced the levels of IL-4, IL-5, IL-13 and IL-17 enhanced the levels of INF-γ in CD4+ T cells, which was rescued by NEAT1 overexpression. Moreover, NEAT1 promoted STAT6 protein expression, whereas NEAT1 had no effect on the expression of STAT6 mRNA. Furthermore, NEAT1 interacted with STAT6, inhibited the ubiquitination of STAT6 in CD4+ T cells. In conclusion, our work has confirmed that NEAT1 promotes STAT6 expression by inhibiting STAT6 ubiquitination, thereby promoting Th2 cell activation. Thus, our work may highlight novel insights into the molecular mechanism of NEAT1 in regulating Th2 cell activation.

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Acknowledgements

This work was funded by the National Natural Science Foundation of China (Grant Number: 8157031013).

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Authors and Affiliations

  1. The Rhinology Department, The First Affiliated Hospital of Zhengzhou University, No. 1 Jianshe East Road, Zhengzhou, 450052, China

    Shuman Huang, Dong Dong, Yaqian Zhang, Zhuo Chen, Jing Geng & Yulin Zhao

  2. Academy of Medical Science, Zhengzhou University, Zhengzhou, 450052, China

    Shuman Huang & Jing Geng

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  1. Shuman Huang
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Correspondence to Yulin Zhao.

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All protocols in this study were authorized by the Ethics Committee of the First Affiliated Hospital of Zhengzhou University (JN.No20180712c0420503 [301]). The volunteers were informed and gave written consent.

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13577_2021_496_MOESM1_ESM.tif

Supplement figure 1 LncRNA NEAT1 was located in nucleus. FISH assay was performed to estimate the subcellular localization of lncRNA NEAT1 in CD4+ T cells. N = 5 (TIF 113 KB)

13577_2021_496_MOESM2_ESM.tif

Supplement figure 2 NEAT1 was silenced in CD4+ T cells. QRT-PCR was performed to assess the expression of NEAT1 in CD4+ T cells following transfection of Si-NEAT1 1 or Si-NEAT1 2. **P < 0.01 vs. Scramble. N = 5 (TIF 166 KB)

13577_2021_496_MOESM3_ESM.tif

Supplement figure 3 STAT6 was silenced in CD4+ T cells. QRT-PCR was performed to assess the expression of STAT6 in CD4+ T cells following transfection of Si-STAT6 1 or Si-STAT6 2. **P < 0.01 vs. Scramble. N = 5 (TIF 162 KB)

13577_2021_496_MOESM4_ESM.tif

Supplement figure 4 NEAT1 overexpression reduced INF-γ production and enhanced IL-17 production in CD4+ T cells. N = 5. ELISA was performed to examine the levels of INF-γ and IL17 in the CD4+ T cells following transfection of pcDNA3.1-NEAT1 or Si-NEAT1 (Si-NEAT1 1). **P < 0.01 vs. Vector; ##P < 0.01 vs. Scramble. N = 5 (TIF 317 KB)

13577_2021_496_MOESM5_ESM.tif

Supplement figure 5 NEAT1 overexpression reduced INF-γ production and enhanced IL-17 production in CD4+ T cells by regulating STAT6. ELISA was performed to examine the levels of INF-γ and IL17 in the CD4+ T cells following transfection of pcDNA3.1-NEAT1 or combined with Si-STAT6 (Si-STAT6 1). ##P < 0.01 vs. Vector + Scramble; $$P < 0.01 vs. Vector + Si-STAT6. N = 5 (TIF 375 KB)

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Huang, S., Dong, D., Zhang, Y. et al. Long non-coding RNA nuclear paraspeckle assembly transcript 1 promotes activation of T helper 2 cells via inhibiting STAT6 ubiquitination. Human Cell 34, 800–807 (2021). https://doi.org/10.1007/s13577-021-00496-1

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