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

Differential Expression of LEF1 Isoforms in Adult Lymphoid and Myeloid Malignancies

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

References

1. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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