Bioinformatics Analysis Integrating Metabolomics of m⁶A RNA Microarray in Intervertebral Disc Degeneration

Abstract

Aim: To explore the potential functions and mechanism of N⁶.methyladenosine (m⁶A) abnormality of RNAs in nucleus pulposus from the intervertebral disc degeneration (IDD). Materials & methods: We performed rat model, m⁶A epitranscriptomic microarray, bioinformatics analysis and metabolomics. Results: In IDD, most of the differentially methylated RNAs showed a significant demethylation situation. The competing endogenous RNA network LOC102555094/miR-431/GSK-3β combining downstream Wnt pathway were identified in bioinformatics analysis. For metabolomics, activation of Wnt pathway led to reprogramming of glucose metabolism and enzyme activation of PKM2. Finally, quantitative real-time PCR and methylated RNA immunoprecipitation coupled with quantitative real-time PCR revealed the positive correlation between demethylation of LOC102555094 and expression of both FTO and ZFP217. Conclusion:LOC102555094 might be demethylated by ZFP217, activating FTO and LOC102555094/miR-431/GSK-3β/Wnt played a crucial role in IDD.

Keywords: :

• bioinformatics analysis
• competing endogenous RNA
• intervertebral disc degeneration
• long noncoding RNA
• microRNA
• N ⁶.methyladenosine

Supplementary data

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Financial #x0026; competing interests disclosure

This study was supported by the National Natural Science Foundation of China (nos. 81972135 and 81401839), the Youth Teacher Program of Sun Yat-sen University (16ykpy38), the Sanming Project of Medicine in Shenzhen (no. SZSM201911002), Funds for Dongguan Social Science #x0026; Technology Development (no. 201750715001441) and the National Natural Science Foundation of China Youth Found (no. 81801907). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations.

Additional information

Funding

This study was supported by the National Natural Science Foundation of China (nos. 81972135 and 81401839), the Youth Teacher Program of Sun Yat-sen University (16ykpy38), the Sanming Project of Medicine in Shenzhen (no. SZSM201911002), Funds for Dongguan Social Science #x0026; Technology Development (no. 201750715001441) and the National Natural Science Foundation of China Youth Found (no. 81801907). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript.