Xiaoshuai Wang1, Ningning Chen1, Zefeng Du2, Zemin Ling3, Penghui Zhang1, Jiaming Yang1, Mohammed Khaleel4, Anthony N Khoury5, Jianwen Li6, Songbo Li6, Haoyang Huang2, Xinwei Zhou2, Yueyin Han2, Fuxin Wei1. 1. Department of Orthopedics, The Seventh Affiliated Hospital of Sun Yat-sen University, No. 628, Zhenyuan Rd, Shenzhen, 518107, China. 2. Department of Clinical Medicine, Zhongshan Medical College of Sun Yat-sen University, No. 74, Zhongshan Er Rd, Guangzhou, 510030, China. 3. Department of Spine Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510030, China. 4. Department of Orthopaedic Surgery, The University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, TX 75390, USA. 5. Hip Preservation Center, Baylor University Medical Center at Dallas, TX 75390, USA. 6. Affiliated Dongguan People's Hospital of Southern Medical University, Dongguan, 523000, China.
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.
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.