Differential Expression of LEF1 Isoforms in Adult Lymphoid and Myeloid Malignancies

Year 2021, , 184 - 188, 08.12.2021

Sinem Fırtına , Özden Hatırnaz Ng , Yücel Erbilgin , İbrahim Celaleddin Haznedaroğlu , Müge Sayitoğlu

https://doi.org/10.26650/experimed.2021.993743

Abstract

Objective: Lymphoid enhancer-binding factor-1 (LEF1) is one of the key regulators of lymphocyte proliferation and its aberrant ex-pression is a prognostic factor for lymphoid or myeloid malignancies. In this study, we focused on the expression of LEF1 isoforms in several hematological malignancies and found tissuespecific dif-ferential expression for the full-length (FL)-LEF1 gene and its tumor suppressor (∆LEF1) variant.

Material and Method: Fifty-three leukemia/lymphoma patients were included in this study. Diagnostic samples of “lymphoid group” patients: Chronic Lymphoblastic Leukemia (CLL) (n=10), B-cell Acute Lymphoblastic Leukemia (B-ALL) (n=9) and “myeloid group” patients: Chronic Myeloblastic Leukemia (CML) (n=12), Acute Myeloid Leukemia (AML) (n=13), and Multiple Myeloma (MM) (n=9) were studied. Healthy bone marrow, peripheral blood cells, and CD34 positive cells were used as controls. Total (T) and FL-LEF1 transcript levels were examined by using quantitative real-time polymerase chain reaction (qRT-PCR). T and FL-LEF1 mRNA ratios were also evaluated for calculation of ∆LEF1.

Results:LEF1 levels were significantly high in lymphoid malignan-cies, but MM and AML patients have decreased LEF1 levels. Al-though CLL patients have high FL-LEF1 levels, the ratio of the T/FL levels was significantly decreased.

Conclusion:LEF1 is a proliferation factor for lymphocytes and not only its differential overexpression but also the ratio of T/FL isoforms seem to accompany leukemia progress.

Keywords

LEF1, alternative splicing, lymphoid, myeloid, leukemia

Supporting Institution

TUBITAK

Project Number

106S112

Thanks

This work was supported and the Scientific and Technological Research Council of Turkey (TÜBİTAK, Project Nos. 106S112). We would like to thank to Suzin Catal Tatonyan for the support of laboratory studies.

Yetişkin Lenfoid ve Miyeloid Malignitelerde LEF1 İzoformlarının Farklı Ekspresyonu

Abstract

Amaç: Lenfositlerin çoğalmasındaki önemli düzenleyicilerden biri olan lenfoid güçlendirici bağlama faktörü-1 (LEF1)’in anormal eks-presyonu, lenfoid veya miyeloid maligniteler için prognostik bir fak-tördür. Bu çalışmada, farklı LEF1 izoformlarının çeşitli hematolojik malignitelerdeki ekspresyonu incelenmiş ve tüm uzunluktaki LEF1(FL-LEF1) anlatımı ile tümör baskılayıcı özelliğe sahip kısa izoformu (∆LEF1) için dokuya özgü farklılıklar tespit edilmiştir.

Gereç ve Yöntem: Çalışmaya 53 yetişkin lösemi/lenfoma hastasının tanı anı örnekleri dahil edilmiştir. Çalışmaya dahil edilen hastalar, “lenfoid grubu”; Kronik Lenfoblastik Lösemi (KLL) (n=10), B hücreli Akut Lenfoblastik Lösemi (B-ALL) (n=9) ve “miyeloid grubu”; Kronik Miyeloblastik Lösemi (KML) (n=12), Akut Miyeloid Lösemi (AML) (n=13) ve Multipl Miyelom (MM) (n=9) gruplarından oluşmaktadır. Sağlıklı kemik iliği, periferik kan hücreleri ve CD34 pozitif hücreler kontrol olarak kullanılmıştır. Total (T) ve FL-LEF1 transkript seviyeleri, gerçek zamanlı kantitatif polimeraz zincir reaksiyonu (qRT-PZR) ile incelenmiştir. T ve FL-LEF1 oranları da ∆LEF1 hesaplaması için değerlendirilmiştir.

Bulgular:LEF1'in lenfoid malignitelerde anlamlı derecede yüksek olduğu, MM ve AML hastalarında ise LEF1 seviyelerinde azalma olduğu görülmüştür. KLL hastalarında FL-LEF1 seviyeleri yüksek olma-sına rağmen, T/FL-LEF1 seviyelerinin önemli ölçüde azaldığı tespit edilmiştir.

Sonuç:LEF1, lenfositler için bir çoğalma faktörüdür ve sadece aşırı ekspresyonu değil, aynı zamanda T/FL-LEF1 izoformlarının oranının da lösemi ilerlemesine eşlik ettiğini düşündürmektedir..

Keywords

LEF1, alternatif kırpılma, lenfoid, miyeloid, lö-semi

Project Number

106S112

References

• 1. Reya T, O'Riordan M, Okamura R, Devaney E, Willert K, Nusse R, et al. Wnt signaling regulates B lymphocyte proliferation through a LEF-1 dependent mechanism. Immunity 2000; 13: 15-24. [CrossRef] google scholar
• 2. Asally M, Yoneda Y. Beta-catenin can act as a nuclear import re-ceptor for its partner transcription factor, lymphocyte enhancer factor-1 (lef-1). Exp Cell Res 2005; 308: 357-63. [CrossRef] google scholar
• 3. Santiago L, Daniels G, Wang D, Deng FM, Lee P. Wnt signaling pathway protein LEF1 in cancer, as a biomarker for prognosis and a target for treatment. Am J Cancer Res 2017; 7: 1389-406. google scholar
• 4. Sutton LA, Ljungström V, Enjuanes A, Cortese D, Skaftason A, Tausch E, et al. Comparative analysis of targeted next-generation sequencing panels for the detection of gene mutations in chronic lymphocytic leukemia: an ERIC multi-center study. Haematologi-ca 2021; 106: 682-91. [CrossRef] google scholar
• 5. Morgan RG, Ridsdale J, Payne M, Heesom KJ, Wilson MC, David-son A, et al. LEF-1 drives aberrant beta-catenin nuclear localiza-tion in myeloid leukemia cells. Haematologica 2019; 104: 1365-77. [CrossRef] google scholar
• 6. Petropoulos K, Arseni N, Schessl C, Stadler CR, Rawat VP, Desh-pande AJ, et al. A novel role for Lef-1, a central transcription me-diator of Wnt signaling, in leukemogenesis. J Exp Med 2008; 205: 515-22. [CrossRef] google scholar
• 7. Hovanes K, Li TW, Munguia JE, Truong T, Milovanovic T, Lawrence Marsh J, et al. Beta-catenin-sensitive isoforms of lymphoid en-hancer factor-1 are selectively expressed in colon cancer. Nat Genet 2001; 28: 53-7. [CrossRef] google scholar
• 8. Atcha FA, Munguia JE, Li TW, Hovanes K, Waterman ML. A new beta-catenin-dependent activation domain in T cell factor. J Biol Chem 2003; 278: 16169-75. [CrossRef] google scholar
• 9. Hovanes K, Li TW, Waterman ML. The human LEF-1 gene contains a promoter preferentially active in lymphocytes and encodes mul-tiple isoforms derived from alternative splicing. Nucleic Acids Res 2000; 28: 1994-2003. [CrossRef] google scholar
• 10. Metzeler KH, Heilmeier B, Edmaier KE, Rawat VP, Dufour A, Dohner K, et al. High expression of lymphoid enhancer-binding factor-1 (LEF1) is a novel favorable prognostic factor in cytogenetically normal acute myeloid leukemia. Blood 2012; 120: 2118-26. [CrossRef] google scholar
• 11. Gutierrez A, Jr., Tschumper RC, Wu X, Shanafelt TD, Eckel-Passow J, Huddleston PM, 3rd, et al. LEF-1 is a prosurvival factor in chronic lymphocytic leukemia and is expressed in the preleukemic state of monoclonal B-cell lymphocytosis. Blood 2010; 116: 2975-83. [CrossRef] google scholar
• 12. Seke Etet PF, Vecchio L, Nwabo Kamdje AH. Interactions between bone marrow stromal microenvironment and B-chronic lympho-cytic leukemia cells: any role for Notch, Wnt and Hh signaling pathways? Cell Signal 2012; 24: 1433-43. [CrossRef] google scholar
• 13. Erbilgin Y, Hatirnaz Ng O, Can I, Firtina S, Kucukcankurt F, Karaman S, et al. Prognostic evidence of LEF1 isoforms in childhood acute lymphoblastic leukemia. Int J Lab Hematol 2021; 43(5): 10931103. [CrossRef] google scholar
• 14. Arber DA, Orazi A, Hasserjian R, Thiele J, Borowitz MJ, Le Beau MM, et al. The 2016 revision to the World Health Organization classi-fication of myeloid neoplasms and acute leukemia. Blood 2016; 127: 2391-405. [CrossRef] google scholar
• 15. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25: 402-8. [CrossRef] google scholar
• 16. Danek P, Kardosova M, Janeckova L, Karkoulia E, Vanickova K, Fa-bisik M, et al. beta-Catenin-TCF/LEF signaling promotes steady-state and emergency granulopoiesis via G-CSF receptor upregu-lation. Blood 2020; 136: 2574-87. [CrossRef] google scholar
• 17. Guo X, Zhang R, Liu J, Li M, Song C, Dovat S, et al. Characterization of LEF1 High Expression and Novel Mutations in Adult Acute Lym-phoblastic Leukemia. PLoS One 2015; 10: e0125429. [CrossRef] google scholar
• 18. Lyapichev KA, Sakhdari A, Khoury JD, O'Malley DP, El Hussein S, Yin CC, et al. Lymphoid enhancer binding factor 1 (LEF1) expression is significantly higher in Hodgkin lymphoma associated with Richter syndrome relative to de novo classic Hodgkin lymphoma. Ann Di-agn Pathol 2020; 49: 151636. [CrossRef] google scholar
• 19. Xu R, Huang X, Li C, Deng C, Li M, Wu P, et al. Bone marrow mes-enchymal stromal cells in chronic myelomonocytic leukaemia: overactivated WNT/beta-catenin signalling by parallel RNA se-quencing and dysfunctional phenotypes. Br J Haematol 2021; 193: 928-40. [CrossRef] google scholar
• 20. Ng OH, Erbilgin Y, Firtina S, Celkan T, Karakas Z, Aydogan G, et al. Deregulated WNT signaling in childhood T-cell acute lymphoblas-tic leukemia. Blood Cancer J 2014; 4: e192. [CrossRef] google scholar
• 21. Fetisov TI, Lesovaya EA, Yakubovskaya MG, Kirsanov KI, Belitsky GA. Alterations in WNT Signaling in Leukemias. Biochemistry (Mosc) 2018; 83: 1448-58. [CrossRef] google scholar
• 22. Jimenez J, Jang GM, Semler BL, Waterman ML. An internal ribo-some entry site mediates translation of lymphoid enhancer fac-tor-1. RNA 2005; 11: 1385-99. [CrossRef] google scholar
• 23. Li TW, Ting JH, Yokoyama NN, Bernstein A, van de Wetering M, Wa-terman ML. Wnt activation and alternative promoter repression of LEF1 in colon cancer. Mol Cell Biol 2006; 26: 5284-99. [CrossRef] google scholar
• 24. Hsu SC, Galceran J, Grosschedl R. Modulation of transcriptional regulation by LEF-1 in response to Wnt-1 signaling and associa-tion with beta-catenin. Mol Cell Biol 1998; 18: 4807-18. [CrossRef] google scholar
• 25. Skokowa J, Cario G, Uenalan M, Schambach A, Germeshausen M, Battmer K, et al. LEF-1 is crucial for neutrophil granulocytopoiesis and its expression is severely reduced in congenital neutropenia. Nat Med 2006; 12: 1191-7. [CrossRef] google scholar
• 26. Menter T, Trivedi P, Ahmad R, Flora R, Dirnhofer S, Tzankov A, et al. Diagnostic Utility of Lymphoid Enhancer Binding Factor 1 Im-munohistochemistry in Small B-Cell Lymphomas. Am J Clin Pathol 2017; 147: 292-300. [CrossRef] google scholar
• 27. Soliman DS, Al-Kuwari E, Siveen KS, Al-Abdulla R, Chandra P, Yassin M, et al. Downregulation of Lymphoid enhancer-binding factor 1 (LEF-1) expression (by immunohistochemistry and/ flow cytometry) in chronic Lymphocytic Leukemia with atypical immunophenotypic and cytologic features. Int J Lab Hematol 2021; 43: 515-25. [CrossRef] google scholar
• 28. Kuhnl A, Gokbuget N, Kaiser M, Schlee C, Stroux A, Burmeister T, et al. Overexpression of LEF1 predicts unfavorable outcome in adult patients with B-precursor acute lymphoblastic leukemia. Blood 2011; 118: 6362-7. [CrossRef] google scholar
• 29. Yokoyama NN, Pate KT, Sprowl S, Waterman ML. A role for YY1 in repression of dominant negative LEF-1 expression in colon can-cer. Nucleic Acids Res 2010; 38: 6375-88. [CrossRef] google scholar
• 30. Masamoto Y, Kurokawa M. Targeting chronic myeloid leukemia stem cells: can transcriptional program be a druggable target for cancers? Stem Cell Investig 2018; 5:10. [CrossRef] google scholar
• 31. Feder K, Edmaier-Schroger K, Rawat VPS, Kirsten N, Metzeler K, Kraus JM, et al. Differences in expression and function of LEF1 iso-forms in normal versus leukemic hematopoiesis. Leukemia 2020; 34: 1027-37. [CrossRef] google scholar