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Extracellular vesicles from thalassemia patients carry iron-containing ferritin and hemichrome that promote cardiac cell proliferation

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Abstract

Extracellular vesicles (EVs) are bioactive, submicron-sized membrane vesicles released from all cell types upon activation or apoptosis. EVs including microparticles (MPs) and exosomes have emerged as important mediators of cell-to-cell communication in both normal and pathological states including thalassemia (thal). However, the role of EVs derived from β-thal patients with iron overload (+ IO) and without iron overload (-IO) on cardiac cells is unclear. We hypothesized plasma EVs in thal patients containing ferritin (iron storage protein) and a denaturated hemoglobin-hemichrome that induce cardiac cell proliferation. The origins and numbers of EVs isolated from plasma of normal, thal (+ IO), and (− IO) patients were compared and determined for their iron and iron-containing proteins along with their effects on cardiac and endothelial cells. Data shows that MPs were originated from many cell sources with marked numbers of platelet origin. Only the number of RBC-derived MPs in thal (+ IO) patients was significantly high when compared to normal controls. Although MPs derived from both normal and thal patients promoted cardiac cell proliferation in a dose-dependent manner, only exosomes from thal patients promoted cardiac cell proliferation compared to the untreated. Moreover, the exosomes from thal (+ IO) potentially induce higher cardiac cell proliferation and angiogenesis in terms of tube number than thal (− IO) and normal controls. Interestingly, ferritin content in the exosomes isolated from thal (+ IO) was higher than that found in the MPs isolated from the same patient. The exosomes of thal patients with higher serum ferritin level also contained greater level of ferritin inside the exosomes. Apart from ferritin, there were trends of increasing hemichrome and iron presented in the plasma EVs and EV-treated H9C2 cells. Findings from this study support the hypothesis that EVs from β-thal patients carry iron-load proteins that leads to the induction of cardiac cell proliferation.

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Acknowledgements

The authors gratefully acknowledge the kind co-operation of thalassemia patients. We thank the primary care physicians and nurses at the Division of Hematology, Siriraj Hospital, for providing us important clinical supports. We would also like to thank Professor Aftab A. Ansari of the Department of Pathology and Laboratory, Emory University School of Medicine, Atlanta, GA, for his valuable comments and suggestions.

Funding

This work was supported by the Thailand Research Fund (TRF) – Distinguished Research Professor Grant, grant number DPG5980001, and the research grant from Faculty of Medicine Siriraj Hospital, Mahidol University. L.K was also supported by Siriraj Chalermprakiat Foundation, Faculty of Medicine Siriraj Hospital, Mahidol University.

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  1. Anyapat Atipimonpat

    Present address: Department of Biochemistry, Faculty of Medical Science, Naresuan University, Phitsanulok, 65000, Thailand

Authors and Affiliations

  1. Graduate Program in Immunology, Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand

    Anyapat Atipimonpat & Kovit Pattanapanyasat

  2. Center of Excellence for Microparticle and Exosome in Diseases, Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand

    Anyapat Atipimonpat, Panjaree Siwaponanan, Kasama Sukapirom & Kovit Pattanapanyasat

  3. Division of Hematology, Department of Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand

    Archrob Khuhapinant

  4. Thalassemia Research Center, Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom, Thailand

    Saovaros Svasti

  5. Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok, Thailand

    Saovaros Svasti

  6. Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand

    Ladawan Khowawisetsut

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Contributions

KP: research idea formation and supervision; AA: performed the experiment and data analysis; AK: patient recruitment and diagnosis; AA, PS, SS, KS, and LK: data analysis; AA and KP: manuscript writing. All authors read and approved the final manuscript.

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Correspondence to Ladawan Khowawisetsut or Kovit Pattanapanyasat.

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All procedures performed in studies involving human participants were in accordance with the 1964 Helsinki declaration and its later amendments or comparable standards. The study was approved by Siriraj Hospital Institutional Review Board (SI-IRB), approval number Si 239/2018. Informed consent was obtained from all individual participants included in the study.

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Atipimonpat, A., Siwaponanan, P., Khuhapinant, A. et al. Extracellular vesicles from thalassemia patients carry iron-containing ferritin and hemichrome that promote cardiac cell proliferation. Ann Hematol 100, 1929–1946 (2021). https://doi.org/10.1007/s00277-021-04567-z

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