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Exploration of microRNA-106b-5p as a therapeutic target in intervertebral disc degeneration: a preclinical study
Dihua Meng1, Weiyou Chen1, Cheng Pan1
1Division of Spine Surgery, The First Affiliated Hospital of Guangxi Medical University, 6 Shuangyong Road, 530021, Nanning, Guangxi Zhuang Autonomous Region, China.
Abstract:
MicroRNA (miRNA) has emerge as a vital regulator in the pathogenesis of intervertebral disc degeneration (IDD). However, miR-106b-5p expression in the human nucleus pulposus (NP) and potential mechanisms remain to be elucidated. In this study, the aim was to verify the potential therapeutic mechanisms of miR-106b-5p for IDD. Key miRNAs were screened for in degenerative and normal human intervertebral disc samples. qRT-PCR and fluorescence in situ hybridization (FISH) were used to verify the miR-106b-5p differential expression. The targeting link between miR-106b-5p and Sirtuin 2 (SIRT2) was identified using the luciferase reporter assay and bioinformatics. Flow cytometry, EdU method, and cell scratching were all performed to determine the NP cell function and IDD models were constructed for in vivo experiments. SIRT2, MMP13, ADAMTS5, Col II, Aggrecan, Ras, ERK1/2, and p-ERK1/2 protein levels were assayed by western blotting. Overexpression of miR-106b-5p in NP cells decreased cell growth, induced apoptosis, hindered extracellular matrix formation, and increased the expression of matrix-degrading enzymes through the SIRT2/MAPK/ERK signaling pathway. Importantly, intradiscal delivery of antagomiR-106b-5p significantly attenuated IDD development. Our findings demonstrate that targeting miR-106b-5p in intervertebral disc has therapeutic effects on IDD.
Insights
MicroRNA-106b-5p (miR-106b-5p) exacerbates intervertebral disc degeneration (IDD) by targeting SIRT2 and activating the MAPK/ERK pathway. Inhibiting miR-106b-5p offers a potential therapeutic strategy for IDD.
Area of Science:
- Biochemistry
- Molecular Biology
- Regenerative Medicine
Background:
- Intervertebral disc degeneration (IDD) is a complex pathological process.
- MicroRNAs (miRNAs) play a crucial role in IDD pathogenesis.
- The specific role and mechanisms of miR-106b-5p in human nucleus pulposus (NP) cells require further investigation.
Purpose of the Study:
- To investigate the expression and function of miR-106b-5p in human NP cells.
- To elucidate the therapeutic potential of targeting miR-106b-5p for IDD.
Main Methods:
- Screening of key miRNAs in degenerative and normal human intervertebral disc samples.
- Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and fluorescence in situ hybridization (FISH) for miR-106b-5p expression analysis.
- Luciferase reporter assay, bioinformatics, flow cytometry, EdU assay, cell scratching, and western blotting to determine molecular mechanisms and cellular functions.
- In vivo IDD models for therapeutic evaluation.
Main Results:
- miR-106b-5p was differentially expressed in degenerative human intervertebral discs.
- Overexpression of miR-106b-5p in NP cells reduced cell proliferation, induced apoptosis, impaired extracellular matrix formation, and increased matrix-degrading enzyme expression via the SIRT2/MAPK/ERK pathway.
- Intradiscal administration of antagomiR-106b-5p effectively attenuated IDD progression in vivo.
Conclusions:
- miR-106b-5p acts as a pro-degenerative factor in IDD by targeting SIRT2 and modulating the SIRT2/MAPK/ERK signaling pathway.
- Targeting miR-106b-5p demonstrates significant therapeutic potential for mitigating intervertebral disc degeneration.
