Identification of characteristic proteins at late-stage erythroid differentiation in vitro

Abstract

The production of red blood cells in vitro, which is useful for basic or clinical research, has been improved. Further optimization of culture protocols may facilitate erythroid differentiation from hematopoietic stem cells to red blood cells. However, the details of erythropoiesis, particularly regarding the behaviors of differentiation-related proteins, remain unclear. Here, we performed erythroid differentiation using two independent bone marrow- or cord blood-derived CD34+ cell sources and identified proteins showing reproducible differential expression in all groups. Notably, most of the proteins expressed at the early stage were downregulated during erythroid differentiation. However, seven proteins showed upregulated expression in both bone marrow cells and cord blood cells. These proteins included alpha-synuclein and selenium-binding protein 1, the roles of which have not been clarified in erythropoiesis. There is a possibility that these factors contribute to erythroid differentiation as they maintained a high expression level. These findings provide a foundation for further mechanistic studies on erythropoiesis.

This is a preview of subscription content, access via your institution.

Fig. 1

References

  1. 1.

    Giarratana MC, Rouard H, Dumont A, et al. Proof of principle for transfusion of in vitro-generated red blood cells. Blood. 2011;118:5071–9. https://doi.org/10.1182/blood-2011-06-362038.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  2. 2.

    Griffiths RE, Kupzig S, Cogan N, Mankelow TJ, et al. Maturing reticulocytes internalize plasma membrane in glycophorin A-containing vesicles that fuse with autophagosomes before exocytosis. Blood. 2012;119:6296–306. https://doi.org/10.1182/blood-2011-09-376475.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  3. 3.

    Bell AJ, Satchwell TJ, Heesom KJ, et al. Protein distribution during human erythroblast enucleation in vitro. PLoS ONE. 2013;8:e60300. https://doi.org/10.1371/journal.pone.0060300.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  4. 4.

    Chu TTT, Sinha A, Malleret B, et al. Quantitative mass spectrometry of human reticulocytes reveal proteome-wide modifications during maturation. Br J Haematol. 2018;180:118–33. https://doi.org/10.1111/bjh.14976.

    CAS  Article  PubMed  Google Scholar 

  5. 5.

    Trakarnsanga K, Wilson MC, Griffiths RE, et al. Qualitative and quantitative comparison of the proteome of erythroid cells differentiated from human iPSCs and adult erythroid cells by multiplex TMT labelling and nanoLC-MS/MS. PLoS ONE. 2014;9:e100874. https://doi.org/10.1371/journal.pone.0100874.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  6. 6.

    Gautier EF, Ducamp S, Leduc M, et al. Comprehensive proteomic analysis of human erythropoiesis. Cell Rep. 2016;16:1470–84. https://doi.org/10.1016/j.celrep.2016.06.085.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  7. 7.

    Masuda T, Saito N, Tomita M, Ishihama Y. Unbiased quantitation of Escherichia coli membrane proteome using phase transfer surfactants. Mol Cell Proteomics. 2009;8:2770–7. https://doi.org/10.1074/mcp.M900240-MCP200.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  8. 8.

    Garel MC, Joulin V, Le Boulch P, et al. Human bisphosphoglycerate mutase Expression in Escherichia coli and use of site-directed mutagenesis in the evaluation of the role of the carboxyl-terminal region in the enzymatic mechanism. J Biol Chem. 1989;264:18966–72.

    CAS  Article  Google Scholar 

  9. 9.

    Joulin V, Garel MC, Le Boulch P, et al. Isolation and characterization of the human 2,3-bisphosphoglycerate mutase gene. J Biol Chem. 1988;263:15785–90.

    CAS  Article  Google Scholar 

  10. 10.

    Pritlove DC, Gu M, Boyd CA, Randeva HS, Vatish M. Novel placental expression of 2,3-bisphosphoglycerate mutase. Placenta. 2006;27:924–7. https://doi.org/10.1016/j.placenta.2005.08.010.

    CAS  Article  PubMed  Google Scholar 

  11. 11.

    Ludwig LS, Lareau CA, Bao EL, Nandakumar SK, Muus C, Ulirsch JC, et al. Transcriptional states and chromatin accessibility underlying human erythropoiesis. Cell Rep. 2019;27:3228–40. https://doi.org/10.1016/j.celrep.2019.05.046.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  12. 12.

    Tong J, Wong H, Guttman M, et al. Brain alpha-synuclein accumulation in multiple system atrophy, Parkinson’s disease and progressive supranuclear palsy: a comparative investigation. Brain. 2010;133:172–88. https://doi.org/10.1093/brain/awp282.

    Article  PubMed  Google Scholar 

  13. 13.

    Araki K, Yagi N, Nakatani R, et al. A small-angle X-ray scattering study of alpha-synuclein from human red blood cells. Sci Rep. 2016;6:30473. https://doi.org/10.1038/srep30473.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  14. 14.

    Araki K, Sugawara K, Hayakawa EH, et al. The localization of α-synuclein in the process of differentiation of human erythroid cells. Int J Hematol. 2018;108:130–8. https://doi.org/10.1007/s12185-018-2457-8.

    CAS  Article  PubMed  Google Scholar 

  15. 15.

    Pol A, Renkema GH, Tangerman A, et al. Mutations in SELENBP1, encoding a novel human methanethiol oxidase, cause extraoral halitosis. Nat Genet. 2018;50:120–9. https://doi.org/10.1038/s41588-017-0006-7.

    CAS  Article  PubMed  Google Scholar 

Download references

Funding

Not applicable.

Author information

Affiliations

Authors

Contributions

KF, TA, RK, YW, and YF conceived and designed the experiments. KF and TA performed the experiments. KF, TA, and RK analyzed the data. YN contributed essential reagents/materials/analysis tools. KF, RK, TA, SM, YF, and MS wrote the paper.

Corresponding author

Correspondence to Ryo Kurita.

Ethics declarations

Conflicts of interest

The authors have no relevant financial or non-financial interests to disclose.

Ethics approval

Approval was obtained from the ethics committee of Japanese Red Cross.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (XLSX 19171 KB)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Funato, K., Abe, T., Kurita, R. et al. Identification of characteristic proteins at late-stage erythroid differentiation in vitro. Human Cell (2021). https://doi.org/10.1007/s13577-021-00503-5

Download citation

Keywords

  • Erythropoiesis
  • CD34+ cell
  • Mass spectrometry
  • Protein identification
  • Differentiation-related factor