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N6-Methyladenosine-modified lncRNA LINREP promotes Glioblastoma progression by recruiting the PTBP1/HuR complex

Cell Death & Differentiation volume 30, pages 54–68 (2023)Cite this article

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Abstract

Glioblastoma multiforme (GBM) is acknowledged as the most aggressive primary brain tumor in adults. It is typically characterized by the high heterogeneity which corresponds to extensive genetic mutations and complex alternative splicing (AS) profiles. Known as a major repressive splicing factor in AS, polypyrimidine tract-binding protein 1 (PTBP1) is involved in the exon skipping events of multiple precursor mRNAs (pre-mRNAs) in GBM. However, precise mechanisms that modulate the expression and activity of PTBP1 remain to be elucidated. In present study, we provided evidences for the role of a long intergenic noncoding RNA (LINREP) implicated in the regulation of PTBP1-induced AS. LINREP interacted with PTBP1 and human antigen R (HuR, ELAVL1) protein complex and protected PTBP1 from the ubiquitin-proteasome degradation. Consequently, a broad spectrum of PTBP1-induced spliced variants was generated by exon skipping, especially for the skipping of reticulon 4 (RTN4) exon 3. Interestingly, LINREP also promoted the dissociation of nuclear UPF1 from PTBP1, which increased the binding of PTBP1 to RTN4 transcripts, thus enhancing the skipping of RTN4 exon 3 to some extent. Besides, HuR recruitment was essential for the stabilization of LINREP via a manner dependent on N6-methyladenosine (m6A) formation and identification. Taken together, our results demonstrated the functional significance of LINREP in human GBM for its dual regulation of PTBP1-induced AS and its m6A modification modality, implicating that HuR/LINREP/PTBP1 axis might serve as a potential therapeutic target for GBM.

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Fig. 1: Identification and characterization of LINREP in GBM.
Fig. 2: LINREP knockdown impaired the proliferation, invasion and migration of GBM cells.
Fig. 3: LINREP was involved in the proteasomal degradation of PTBP1.
Fig. 4: LINREP modulated AS of RTN4 via interaction with PTBP1.
Fig. 5: LINREP regulated the interaction between PTBP1 and UPF1.
Fig. 6: LINREP interacted with PTBP1 to affect the proliferation, invasion and migration of GBM cells.
Fig. 7: HuR enhanced LINREP stability via an m6A-dependent manner.
Fig. 8: A schematic model for the mechanisms of LINREP in the development of GBM.

Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

We thank Mengqi Wang, Xiaowei Chen and Wenjie Zhu (Shandong University) for providing advice and technical assistance.

Funding

This work was supported by Natural Science Foundation of China (81972340, 81871196 and 81471517), Key project of Shandong Provincial Natural Science Foundation (ZR202010300086), Science and Technology Project of Jinan city (201907048), Key Projects of Natural Science Foundation of Jiangxi Province (20192ACB20011), Shandong Province Key Research and Development Program (2019GSF107046) and Academic promotion program of Shandong First Medical University (2019LJ005).

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Author notes

  1. These authors contributed equally: Xiaoshuai Ji, Zihao Liu, Jiajia Gao.

Authors and Affiliations

  1. Department of Neurosurgery, Shandong Provincial Qianfoshan Hospital, Cheeloo College of Medicine, Shandong University, Jinan, 250014, China

    Xiaoshuai Ji, Jiajia Gao, Wenqing Liu, Fenglin Zhang & Tao Xin

  2. Department of Neurosurgery, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinan, 250021, China

    Zihao Liu & Dong He

  3. Department of Otolaryngology, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinan, 250021, China

    Xin Bing

  4. Department of Neurosurgery, Shandong Provincial Qianfoshan Hospital, Shandong First Medical University & Shandong Academy of Medical Sciences, Shandong Medicine and Health Key Laboratory of Neurosurgery, Jinan, 250014, China

    Yunda Wang, Xianyong Yin, Min Han & Tao Xin

  5. Department of Histology and Embryology, School of Basic Medical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, 250012, China

    Yanbang Wei, Zixiao Wang & Qian Liu

  6. Department of Neurosurgery, Jiangxi Provincial People’s Hospital Affiliated to Nanchang University, Nanchang, 330006, Jiangxi, China

    Xiangdong Lu & Tao Xin

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Contributions

TX and QL designed the project. XJ, ZL and JG conducted the experiments. XJ, ZL and DH participated in data analysis and figure preparation. XJ, ZL and JG wrote the manuscript. XB, DH, WL and YW contributed to the clinical sample collection and pathological analysis. YW, YX and FZ helped with the animal study. MH, XL, ZW and XB extracted the information from databases. TX, QL and XB reviewed the manuscript. All authors read and approved the final manuscript.

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Correspondence to Qian Liu or Tao Xin.

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The authors declare no competing interests.

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The research was approved by the Research Ethics Committee of Shandong University and performed in accordance with the ethical guidelines of World Medical Association Declaration of Helsinki. Written informed consent was obtained from all patients. All of the animal experiments were approved by the Animal Care and Use Committee of Shandong Provincial Qianfoshan Hospital, Shandong University.

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Ji, X., Liu, Z., Gao, J. et al. N6-Methyladenosine-modified lncRNA LINREP promotes Glioblastoma progression by recruiting the PTBP1/HuR complex. Cell Death Differ 30, 54–68 (2023). https://doi.org/10.1038/s41418-022-01045-5

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