Tandem Mass Tag-Based Proteomic Analysis of Normal and Degenerated Human Intervertebral Discs

Abstract

Background

Intervertebral disc degeneration (IVDD) is the main cause of low back pain (LBP), but the specific regulatory factors, pathways and specific molecular mechanisms remain unclear.

Methods

We identified and quantitatively analyzed Pfirrmann Grade II (n=3) and Pfirrmann Grade IV (n=3) pulposus samples via MRI. The differential abundance of proteins in the samples was determined and quantitatively analyzed by relative and absolute quantitative analysis of the isotope marker levels combined with the liquid chromatography–tandem mass spectrometry (LC‒MSMS/MS).

Results

A total of 70 proteins (30 significantly increased proteins (> 1.2-fold change) and 40 significantly decreased proteins (< 0.8-fold change)) showed different levels among the groups. Kyoto Encyclopedia of Genes and Genomes and Gene Ontology (GO) enrichment analyses and Western blot analysis showed that CYCS, RAC1, and PSMD14 may play important roles in IVDD and that Epstein‒Barr virus infection, viral myocarditis, colorectal cancer, nonalcoholic fatty liver disease (NAFLD) and amyotrophic lateral sclerosis (ALS) are the main pathways involved in IVDD.

Conclusion

CYCS, RAC1 and PSMD14 may play important roles in IVDD, and Epstein‒Barr virus infection, viral myocarditis, colorectal cancer, NAFLD and ALS may be the main pathways involved in IVDD.

Keywords:

• TMT
• LC‒MS/MS
• proteomics
• low back pain
• nucleus pulposus

Data Sharing Statement

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

Ethics Approval and Consent to Participate

All treatments used in this study were performed according to relevant guidelines and rules, and informed consent was obtained from all the participants. The ethics committee of Zhejiang Hospital approved the study (Approval NO.:2022.156K). We followed the Declaration of Helsinki guidelines.

Acknowledgement

The authors thank Jun-Cai Ye for his contribution to Figure 9.

Author Contributions

All authors contributed to data analysis, drafting or revising the article, have agreed on the journal to which the article will be submitted, gave final approval of the version to be published, and agree to be accountable for all aspects of the work. Yang-Fu and Xiao-Qin Huang are co-first authors and contributed eqally to this article.

Disclosure

The authors declare that they have no competing interests in this work.

Additional information

Funding

There is no funding to report.