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Quantitative proteomics analysis of sporadic parathyroid adenoma tissue samples

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Abstract

Objective

Molecular pathogenesis of parathyroid tumors is incompletely understood. Identification of novel molecules and understanding their role in parathyroid tumorigenesis by proteomics approach would be informative with potential clinical implications.

Method

Adenomatous (n = 5) and normal (n = 2) parathyroid tissue lysates were analyzed for protein profile by LC–MS/MS method and the proteins were classified using bioinformatics tools such as PANTHER and toppfun functional enrichment tool. Identified proteins were further validated by western blotting and qRT-PCR (n = 20).

Result

Comparative proteomics analysis revealed that a total of 206 proteins (74 upregulated and 132 downregulated) were differentially expressed (≥ twofold change) in adenomas. Bioinformatics analysis revealed that 48 proteins were associated with plasma membrane, 49 with macromolecular complex, 39 were cytoplasm, 38 were organelle related, 21 were cell junction and 10 were extracellular proteins. These proteins belonged to a diverse protein family such as enzymes, transcription factors, cell signalling, cell adhesion, cytoskeleton proteins, receptors, and calcium-binding proteins. The major biological processes predicted for the proteins were a cellular, metabolic and developmental process, cellular localization, and biological regulation. The differentially expressed proteins were found to be associated with MAPK, phospholipase C (PLC) and phosphatidylinositol (PI) signalling pathways, and with chromatin organization. Western blot and qRT-PCR analysis of three proteins (DNAJC2, ACO2, and PRDX2) validated the LC–MS/MS findings.

Conclusion

This exploratory study demonstrates the feasibility of proteomics approach in finding the dysregulated proteins in benign parathyroid adenomas, and our preliminary results suggest that MAPK, PLC and PI signalling pathways and chromatin organization are involved in parathyroid tumorigenesis.

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Funding

The study was partially funded by Indian Council of Medical Research (ICMR), New Delhi (IRIS Id 2011-15750). AA is grateful to University Grants Commission (UGC), New Delhi, India, for providing the research fellowship. PS and GK are thankful to Indian Council of Medical Research, New Delhi, India, for fellowship.

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Authors and Affiliations

  1. Department of Endocrinology, Postgraduate Institute of Medical Education and Research (PGIMER), Room No. 2, 4th Floor Block-F, Nehru Hospital, PGIMER, Chandigarh, 160012, India

    A. K. Arya, S. K. Bhadada, P. Singh & G. Kaur

  2. Department of General Surgery, PGIMER, Chandigarh, India

    D. Dahiya & A. Behera

  3. Department of Biochemistry, PGIMER, Chandigarh, India

    S. Sharma

  4. Department of Histopathology, PGIMER, Chandigarh, India

    U. N. Saikia

  5. Bone and Mineral Research Laboratory, Henry Ford Hospital, Detroit, USA

    S. D. Rao

  6. Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA

    M. Bhasin

Authors
  1. A. K. Arya
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  2. S. K. Bhadada
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  3. P. Singh
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  4. D. Dahiya
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  5. G. Kaur
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  6. S. Sharma
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  7. U. N. Saikia
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  8. A. Behera
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  9. S. D. Rao
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  10. M. Bhasin
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Contributions

AA, PS and GK performed the molecular experiments. AA, SB, and SS analyzed the data. SB recruited the subjects, DD, and AB performed the surgery and US performed the histopathological analysis of the tumors. AA, SB, and SDR conceptualized the idea and wrote the manuscript. SDR and MB critically reviewed and edited the manuscript. All authors read and approved the submitted version of the manuscript.

Corresponding author

Correspondence to S. K. Bhadada.

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Conflict of interest

The authors exclude any conflict of interests.

Ethical approval

The research protocol was approved by Institutional ethics committee, PGIMER Chandigarh. The study was conducted in accordance with approved protocol.

Informed consent

Informed written consent was taken from each subject prior to the recruitment.

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Arya, A.K., Bhadada, S.K., Singh, P. et al. Quantitative proteomics analysis of sporadic parathyroid adenoma tissue samples. J Endocrinol Invest 42, 577–590 (2019). https://doi.org/10.1007/s40618-018-0958-1

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  • DOI: https://doi.org/10.1007/s40618-018-0958-1

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