Expression of MicroRNA-155 in Thalassemic Erythropoiesis

Overview

Journal PeerJ

Specialties Biology
Environmental Health
General Medicine

Date 2024 Sep 24

PMID 39314840

Authors

Tipparat Penglong

Nuttanan Pholngam

Nasra Tehyoh

Natta Tansila

Hansuk Buncherd

Supinya Thanapongpichat

Kanitta Srinoun

Affiliations

Soon will be listed here.

Abstract

Background: Ineffective erythropoiesis (IE) is the primary cause of anemia and associated pathologies in -thalassemia. The characterization of IE is imbalance of erythroid proliferation and differentiation, resulting in increased erythroblast proliferation that fails to differentiate and gives rise to enucleate RBCs. MicroRNAs (miRs) are known to play important roles in hematopoiesis. miR-155 is a multifunctional molecule involved in both normal and pathological hematopoiesis, and its upregulation is observed in patients with -thalassemia/HbE. However, the expression and function of miR-155, especially in -thalassemia, have not yet been explored.

Methods: To study miR-155 expression in thalassemia, erythroblast subpopulations, CD45-CD71Ter-119 and CD45-CD71Ter-119 were collected from thalassemic bone marrow. Additionally, a two-phase culture of mouse bone marrow erythroid progenitor cells was performed. Expression of miR-155 and predicted mRNA target genes, , and , were determined by quantitative reverse transcription (qRT)-polymerase chain reaction (PCR) and normalized to small nucleolar RNA (snoRNA) 202 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), respectively. To investigate the effect of miR-155 expression, erythroblasts were transfected with miR-inhibitor and -mimic in order to elevate and eliminate miR-155 expression, respectively. Erythroid cell differentiation was evaluated by Wright-Giemsa staining and flow cytometry.

Results: miR-155 was upregulated, both and , during erythropoiesis in -thalassemic mice. Our study revealed that gain- and loss of function of miR-155 were involved in erythroid proliferation and differentiation, and augmented proliferation and differentiation of thalassemic mouse erythroblasts may be associated with miR-155 upregulation. miR-155 upregulation in -thalassemic mice significantly increased the percentage of basophilic and polychromatic erythroblasts. Conversely, a significant decrease in percentage of basophilic and polychromatic erythroblasts was observed in -thalassemic mice transfected with anti-miR-155 inhibitor. We also examined the mRNA targets (, and ) of miR-155, which indicated that is a valid target gene of miR-155 that regulates erythroid differentiation.

Conclusion: miR-155 regulates IE in -thalassemia via expression controlling erythroblast proliferation and differentiation.

References

Jayapal S, Lee K, Ji P, Kaldis P, Lim B, Lodish H . Down-regulation of Myc is essential for terminal erythroid maturation. J Biol Chem. 2010; 285(51):40252-65. PMC: 3001006. DOI: 10.1074/jbc.M110.181073. View

Libani I, Guy E, Melchiori L, Schiro R, Ramos P, Breda L . Decreased differentiation of erythroid cells exacerbates ineffective erythropoiesis in beta-thalassemia. Blood. 2008; 112(3):875-85. PMC: 2481539. DOI: 10.1182/blood-2007-12-126938. View

Penglong T, Saensuwanna A, Pholngam N, Tansila N, Buncherd H, Srinoun K . BACH1 regulates erythrophagocytosis and iron-recycling in β-thalassemia. Genes Cells. 2022; 28(3):211-225. DOI: 10.1111/gtc.13004. View

Liu Y, Pop R, Sadegh C, Brugnara C, Haase V, Socolovsky M . Suppression of Fas-FasL coexpression by erythropoietin mediates erythroblast expansion during the erythropoietic stress response in vivo. Blood. 2006; 108(1):123-33. PMC: 1895827. DOI: 10.1182/blood-2005-11-4458. View

Edalati Fathabad M, Karimipoor M, Alizadeh S, Abdoli A, Atashi A, Sayadi M . miR-155 effectively induces apoptosis in K562 Philadelphia positive cell line through upregulation of p27kip1. Bioimpacts. 2017; 7(2):109-114. PMC: 5524985. DOI: 10.15171/bi.2017.14. View

Undi R, Kandi R, Gutti R . MicroRNAs as Haematopoiesis Regulators. Adv Hematol. 2014; 2013:695754. PMC: 3884629. DOI: 10.1155/2013/695754. View

Sarakul O, Vattanaviboon P, Tanaka Y, Fucharoen S, Abe Y, Svasti S . Enhanced erythroid cell differentiation in hypoxic condition is in part contributed by miR-210. Blood Cells Mol Dis. 2013; 51(2):98-103. DOI: 10.1016/j.bcmd.2013.03.005. View

Siwaponanan P, Fucharoen S, Sirankapracha P, Winichagoon P, Umemura T, Svasti S . Elevated levels of miR-210 correlate with anemia in β-thalassemia/HbE patients. Int J Hematol. 2016; 104(3):338-43. DOI: 10.1007/s12185-016-2032-0. View

Goupille O, Penglong T, Lefevre C, Granger M, Kadri Z, Fucharoen S . BET bromodomain inhibition rescues erythropoietin differentiation of human erythroleukemia cell line UT7. Biochem Biophys Res Commun. 2012; 429(1-2):1-5. DOI: 10.1016/j.bbrc.2012.10.112. View

10.

May A, Forrester L . The erythroblastic island niche: modeling in health, stress, and disease. Exp Hematol. 2020; 91:10-21. DOI: 10.1016/j.exphem.2020.09.185. View

11.

Sun S, Sun P, Wang C, Sun T . Downregulation of microRNA-155 accelerates cell growth and invasion by targeting c-myc in human gastric carcinoma cells. Oncol Rep. 2014; 32(3):951-6. DOI: 10.3892/or.2014.3288. View

12.

Srinoun K, Nopparatana C, Wongchanchailert M, Fucharoen S . MiR-155 enhances phagocytic activity of β-thalassemia/HbE monocytes via targeting of BACH1. Int J Hematol. 2017; 106(5):638-647. DOI: 10.1007/s12185-017-2291-4. View

13.

Leecharoenkiat K, Tanaka Y, Harada Y, Chaichompoo P, Sarakul O, Abe Y . Plasma microRNA-451 as a novel hemolytic marker for β0-thalassemia/HbE disease. Mol Med Rep. 2017; 15(5):2495-2502. PMC: 5428399. DOI: 10.3892/mmr.2017.6326. View

14.

Chen K, Liu J, Heck S, Chasis J, An X, Mohandas N . Resolving the distinct stages in erythroid differentiation based on dynamic changes in membrane protein expression during erythropoiesis. Proc Natl Acad Sci U S A. 2009; 106(41):17413-8. PMC: 2762680. DOI: 10.1073/pnas.0909296106. View

15.

Taher A, Weatherall D, Cappellini M . Thalassaemia. Lancet. 2017; 391(10116):155-167. DOI: 10.1016/S0140-6736(17)31822-6. View

16.

Pootrakul P, Sirankapracha P, Hemsorach S, Moungsub W, Kumbunlue R, Piangitjagum A . A correlation of erythrokinetics, ineffective erythropoiesis, and erythroid precursor apoptosis in thai patients with thalassemia. Blood. 2000; 96(7):2606-12. View

17.

Georgantas 3rd R, Hildreth R, Morisot S, Alder J, Liu C, Heimfeld S . CD34+ hematopoietic stem-progenitor cell microRNA expression and function: a circuit diagram of differentiation control. Proc Natl Acad Sci U S A. 2007; 104(8):2750-5. PMC: 1796783. DOI: 10.1073/pnas.0610983104. View

18.

Das S, Das S, Byram P, Rahaman M, Dolai T, Chatterjee A . MicroRNA expression patterns in HbE/β-thalassemia patients: The passwords to unlock fetal hemoglobin expression in β-hemoglobinopathies. Blood Cells Mol Dis. 2020; 87:102523. DOI: 10.1016/j.bcmd.2020.102523. View

19.

Hoffman B, Liebermann D . Apoptotic signaling by c-MYC. Oncogene. 2008; 27(50):6462-72. DOI: 10.1038/onc.2008.312. View

20.

Rund D, Rachmilewitz E . Beta-thalassemia. N Engl J Med. 2005; 353(11):1135-46. DOI: 10.1056/NEJMra050436. View