Top

Published in:

Open Access 01-12-2018 | Research

NPM1 alternative transcripts are upregulated in acute myeloid and lymphoblastic leukemia and their expression level affects patient outcome

Authors: Luiza Handschuh, Pawel Wojciechowski, Maciej Kazmierczak, Malgorzata Marcinkowska-Swojak, Magdalena Luczak, Krzysztof Lewandowski, Mieczyslaw Komarnicki, Jacek Blazewicz, Marek Figlerowicz, Piotr Kozlowski

Published in: Journal of Translational Medicine | Issue 1/2018

Abstract

Background

Expression of the NPM1 gene, encoding nucleophosmin, is upregulated in cancers. Although more than ten NPM1 transcripts are known, the reports were usually limited to one predominant transcript. In leukemia, the NPM1 expression has not been widely studied so far. In acute myeloid leukemia (AML), the mutational status of the gene seems to play a pivotal role in carcinogenesis. Therefore, the aim of the study was to quantify alternative NPM1 transcripts in two types of acute leukemia, AML and ALL (acute lymphoblastic leukemia).

Methods

Using droplet digital PCR, we analyzed the levels of three protein-coding NPM1 transcripts in 66 samples collected from AML and ALL patients and 16 control samples. Using RNA-seq, we detected 8 additional NPM1 transcripts, including non-coding splice variants with retained introns. For data analysis, Welch two sample t-test, Pearson’s correlation and Kaplan–Meier analysis were applied.

Results

The levels of the particular NPM1 transcripts were significantly different but highly correlated with each other in both leukemia and control samples. Transcript NPM1.1, encoding the longest protein (294 aa), had the highest level of accumulation and was one of the most abundant transcripts in the cell. Comparing to NPM1.1, the levels of the NPM1.2 and NPM1.3 transcripts, encoding a 265-aa and 259-aa proteins, were 30 and 3 times lower, respectively. All three NPM1 transcripts were proportionally upregulated in both types of leukemia compared to control samples. In AML, the levels of NPM1 transcripts decreased in complete remission and increased again with relapse of the disease. Low levels of NPM1.1 and NPM1.3 were associated with better prognosis. The contribution of non-coding transcripts to the total level of NPM1 gene seemed to be marginal, except for one short 5-end transcript accumulated at high levels in AML and control cells. Aberrant proportions of particular NPM1 splice variants could be linked to abnormal expression of genes encoding alternative splicing factors.

Conclusions

The levels of the studied NPM1 transcripts were different but highly correlated with each other. Their upregulation in AML and ALL, decrease after therapy and association with patient outcome suggests the involvement of elevated NPM1 expression in the acute leukemia pathogenesis.

Available only for authorised users

Falini B. Acute myeloid leukemia with mutated nucleophosmin (NPM1): molecular, pathological, and clinical features. Cancer Treat Res. 2010;145:149–68. https://doi.org/10.1007/978-0-387-69259-3_9.CrossRefPubMed

Lindström MS. NPM1/B23: a multifunctional chaperone in ribosome biogenesis and chromatin remodeling. Biochem Res Int. 2011;2011:195209. https://doi.org/10.1155/2011/195209.CrossRefPubMed

Falini B, Nicoletti I, Martelli MF, Mecucci C. Acute myeloid leukemia carrying cytoplasmic/mutated nucleophosmin (NPMc + AML): biologic and clinical features. Blood. 2007;109:874–85. https://doi.org/10.1182/blood-2006-07-012252.CrossRefPubMed

Chen W, Rassidakis GZ, Medeiros LJ. Nucleophosmin gene mutations in acute myeloid leukemia. Arch Pathol Lab Med. 2006;130:1687–92. https://doi.org/10.1043/1543-2165(2006)130[1687:NGMIAM]2.0.CO;2.PubMedCrossRef

Falini B, Mecucci C, Tiacci E, Alcalay M, Rosati R, Pasqualucci L, La Starza R, Diverio D, Colombo E, Santucci A, Bigerna B, Pacini R, Pucciarini A, et al. Cytoplasmic nucleophosmin in acute myelogenous leukemia with a normal karyotype. N Engl J Med. 2005;352:254–66. https://doi.org/10.1056/NEJMoa041974.CrossRefPubMed

Alcalay M, Tiacci E, Bergomas R, Bigerna B, Venturini E, Minardi SP, Meani N, Diverio D, Bernard L, Tizzoni L, Volorio S, Luzi L, Colombo E, et al. Acute myeloid leukemia bearing cytoplasmic nucleophosmin (NPMc + AML) shows a distinct gene expression profile characterized by up-regulation of genes involved in stem-cell maintenance. Blood. 2005;106:899–902. https://doi.org/10.1182/blood-2005-02-0560.CrossRefPubMed

Becker H, Marcucci G, Maharry K, Radmacher MD, Mrózek K, Margeson D, Whitman SP, Wu Y-Z, Schwind S, Paschka P, Powell BL, Carter TH, Kolitz JE, et al. Favorable prognostic impact of NPM1 mutations in older patients with cytogenetically normal de novo acute myeloid leukemia and associated gene- and microRNA-expression signatures: a Cancer and Leukemia Group B study. J Clin Oncol. 2010;28:596–604. https://doi.org/10.1200/JCO.2009.25.1496.CrossRefPubMed

Haferlach C, Mecucci C, Schnittger S, Kohlmann A, Mancini M, Cuneo A, Testoni N, Rege-Cambrin G, Santucci A, Vignetti M, Fazi P, Martelli MP, Haferlach T, et al. AML with mutated NPM1 carrying a normal or aberrant karyotype show overlapping biologic, pathologic, immunophenotypic, and prognostic features. Blood. 2009;114:3024–32. https://doi.org/10.1182/blood-2009-01-197871.CrossRefPubMed

Alpermann T, Kern W, Schnittger S, Schmid C, Kreuzer K-A, Serve H, Pielken HJ, Schmitz N, Wendtner C-M, Haferlach C, Haferlach T. Evaluation of the proposed reporting system of the European LeukemiaNet and recommendations for prognosis of acute myeloid leukemia. Leuk Res. 2013;37:197–200. https://doi.org/10.1016/j.leukres.2012.11.008.CrossRefPubMed

10.

Döhner K, Schlenk RF, Habdank M, Scholl C, Rücker FG, Corbacioglu A, Bullinger L, Fröhling S, Döhner H. Mutant nucleophosmin (NPM1) predicts favorable prognosis in younger adults with acute myeloid leukemia and normal cytogenetics: interaction with other gene mutations. Blood. 2005;106:3740–6. https://doi.org/10.1182/blood-2005-05-2164.CrossRefPubMed

11.

Schnittger S, Schoch C, Kern W, Mecucci C, Tschulik C, Martelli MF, Haferlach T, Hiddemann W, Falini B. Nucleophosmin gene mutations are predictors of favorable prognosis in acute myelogenous leukemia with a normal karyotype. Blood. 2005;106:3733–9. https://doi.org/10.1182/blood-2005-06-2248.CrossRefPubMed

12.

Arber DA, Orazi A, Hasserjian R, Thiele J, Borowitz MJ, Le Beau MM, Bloomfield CD, Cazzola M, Vardiman JW. The 2016 revision to the World Health Organization classification of myeloid neoplasms and acute leukemia. Blood. 2016;127:2391–405. https://doi.org/10.1182/blood-2016-03-643544.CrossRefPubMed

13.

Vardiman JW, Thiele J, Arber DA, Brunning RD, Borowitz MJ, Porwit A, Harris NL, Le Beau MM, Hellström-Lindberg E, Tefferi A, Bloomfield CD. The 2008 revision of the World Health Organization (WHO) classification of myeloid neoplasms and acute leukemia: rationale and important changes. Blood. 2009;114:937–51. https://doi.org/10.1182/blood-2009-03-209262.CrossRefPubMed

14.

Noguera NI, Ammatuna E, Zangrilli D, Lavorgna S, Divona M, Buccisano F, Amadori S, Mecucci C, Falini B, Lo-Coco F. Simultaneous detection of NPM1 and FLT3-ITD mutations by capillary electrophoresis in acute myeloid leukemia. Leukemia. 2005;19:1479–82. https://doi.org/10.1038/sj.leu.2403846.CrossRefPubMed

15.

Scholl S, Mügge L-O, Landt O, Loncarevic IF, Kunert C, Clement JH, Höffken K. Rapid screening and sensitive detection of NPM1 (nucleophosmin) exon 12 mutations in acute myeloid leukaemia. Leuk Res. 2007;31:1205–11. https://doi.org/10.1016/j.leukres.2006.12.011.CrossRefPubMed

16.

Gruszka AM, Lavorgna S, Consalvo MI, Ottone T, Martinelli C, Cinquanta M, Ossolengo G, Pruneri G, Buccisano F, Divona M, Cedrone M, Ammatuna E, Venditti A, et al. A monoclonal antibody against mutated nucleophosmin 1 for the molecular diagnosis of acute myeloid leukemias. Blood. 2010;116:2096–102. https://doi.org/10.1182/blood-2010-01-266908.CrossRefPubMed

17.

Oppliger Leibundgut E, Porret NA, Bienz Muggli M, Baumgartner H, Dahlhaus M, Baerlocher GM. Rapid and highly specific screening for NPM1 mutations in acute myeloid leukemia. Ann Hematol. 2013;92:173–7. https://doi.org/10.1007/s00277-012-1617-9.CrossRefPubMed

18.

Marcinkowska-Swojak M, Handschuh L, Wojciechowski P, Goralski M, Tomaszewski K, Kazmierczak M, Lewandowski K, Komarnicki M, Blazewicz J, Figlerowicz M, Kozlowski P. Simultaneous detection of mutations and copy number variation of NPM1 in the acute myeloid leukemia using multiplex ligation-dependent probe amplification. Mutat Res. 2016;786:14–26. https://doi.org/10.1016/j.mrfmmm.2016.02.001.CrossRefPubMed

19.

Londero AP, Orsaria M, Tell G, Marzinotto S, Capodicasa V, Poletto M, Vascotto C, Sacco C, Mariuzzi L. Expression and prognostic significance of APE1/Ref-1 and NPM1 proteins in high-grade ovarian serous cancer. Am J Clin Pathol. 2014;141:404–14. https://doi.org/10.1309/AJCPIDKDLSGE26CX.CrossRefPubMed

20.

Loubeau G, Boudra R, Maquaire S, Lours-Calet C, Beaudoin C, Verrelle P, Morel L. NPM1 silencing reduces tumour growth and MAPK signalling in prostate cancer cells. PLoS ONE. 2014;9:e96293. https://doi.org/10.1371/journal.pone.0096293.CrossRefPubMedPubMedCentral

21.

Liu Y, Zhang F, Zhang X-F, Qi L-S, Yang L, Guo H, Zhang N. Expression of nucleophosmin/NPM1 correlates with migration and invasiveness of colon cancer cells. J Biomed Sci. 2012;19:53. https://doi.org/10.1186/1423-0127-19-53.CrossRefPubMedPubMedCentral

22.

Wong JCT, Hasan MR, Rahman M, Yu AC, Chan SK, Schaeffer DF, Kennecke HF, Lim HJ, Owen D, Tai IT. Nucleophosmin 1, upregulated in adenomas and cancers of the colon, inhibits p53-mediated cellular senescence. Int J Cancer J Int Cancer. 2013;133:1567–77. https://doi.org/10.1002/ijc.28180.CrossRef

23.

Tsui K, Juang H, Lee T, Chang P, Chen C, Yung BY. Association of nucleophosmin/B23 with bladder cancer recurrence based on immunohistochemical assessment in clinical samples. Acta Pharmacol Sin. 2008;29:364–70. https://doi.org/10.1111/j.1745-7254.2008.00747.x.CrossRefPubMed

24.

Pianta A, Puppin C, Franzoni A, Fabbro D, Di Loreto C, Bulotta S, Deganuto M, Paron I, Tell G, Puxeddu E, Filetti S, Russo D, Damante G. Nucleophosmin is overexpressed in thyroid tumors. Biochem Biophys Res Commun. 2010;397:499–504. https://doi.org/10.1016/j.bbrc.2010.05.142.CrossRefPubMed

25.

Kim CK, Nguyen TLX, Lee SB, Park SB, Lee K-H, Cho S-W, Ahn J-Y. Akt2 and nucleophosmin/B23 function as an oncogenic unit in human lung cancer cells. Exp Cell Res. 2011;317:966–75. https://doi.org/10.1016/j.yexcr.2010.12.013.CrossRefPubMed

26.

Chan WY, Liu QR, Borjigin J, Busch H, Rennert OM, Tease LA, Chan PK. Characterization of the cDNA encoding human nucleophosmin and studies of its role in normal and abnormal growth. Biochemistry. 1989;28:1033–9.CrossRefPubMed

27.

Yun J-P, Miao J, Chen GG, Tian Q-H, Zhang C-Q, Xiang J, Fu J, Lai PBS. Increased expression of nucleophosmin/B23 in hepatocellular carcinoma and correlation with clinicopathological parameters. Br J Cancer. 2007;96:477–84. https://doi.org/10.1038/sj.bjc.6603574.CrossRefPubMedPubMedCentral

28.

Dergunova NN, Bulycheva TI, Artemenko EG, Shpakova AP, Pegova AN, Gemjian EG, Dudnik OA, Zatsepina OV, Malashenko OS. A major nucleolar protein B23 as a marker of proliferation activity of human peripheral lymphocytes. Immunol Lett. 2002;83:67–72.CrossRefPubMed

29.

Kondo T, Minamino N, Nagamura-Inoue T, Matsumoto M, Taniguchi T, Tanaka N. Identification and characterization of nucleophosmin/B23/numatrin which binds the anti-oncogenic transcription factor IRF-1 and manifests oncogenic activity. Oncogene. 1997;15:1275–81. https://doi.org/10.1038/sj.onc.1201286.CrossRefPubMed

30.

Handschuh L, Kaźmierczak M, Milewski MC, Góralski M, Łuczak M, Wojtaszewska M, Uszczyńska-Ratajczak B, Lewandowski K, Komarnicki M, Figlerowicz M. Gene expression profiling of acute myeloid leukemia samples from adult patients with AML-M1 and -M2 through boutique microarrays, real-time PCR and droplet digital PCR. Int J Oncol. 2018;52:656–78. https://doi.org/10.3892/ijo.2017.4233.PubMedCrossRef

31.

Zajac M, Dolnik A, Stasiak G, Zaleska J, Kielbus M, Czapinski J, Schunn M, Correa SC, Glodkowska-Mrowka E, Sundaram RC, Jankowska-Lecka O, Schlenk RF, Döhner H, et al. Analysis of NPM1 splice variants reveals differential expression patterns of prognostic value in acute myeloid leukemia. Oncotarget. 2017;8:95163–75. https://doi.org/10.18632/oncotarget.19871.CrossRef

32.

Hisaoka M, Nagata K, Okuwaki M. Intrinsically disordered regions of nucleophosmin/B23 regulate its RNA binding activity through their inter- and intra-molecular association. Nucleic Acids Res. 2014;42:1180–95. https://doi.org/10.1093/nar/gkt897.CrossRefPubMed

33.

Box JK, Paquet N, Adams MN, Boucher D, Bolderson E, O’Byrne KJ, Richard DJ. Nucleophosmin: from structure and function to disease development. BMC Mol Biol. 2016;17:19. https://doi.org/10.1186/s12867-016-0073-9.CrossRefPubMedPubMedCentral

34.

Döhner H, Estey EH, Amadori S, Appelbaum FR, Büchner T, Burnett AK, Dombret H, Fenaux P, Grimwade D, Larson RA, Lo-Coco F, Naoe T, Niederwieser D, et al. Diagnosis and management of acute myeloid leukemia in adults: recommendations from an international expert panel, on behalf of the European LeukemiaNet. Blood. 2010;115:453–74. https://doi.org/10.1182/blood-2009-07-235358.CrossRefPubMed

35.

Hindson CM, Chevillet JR, Briggs HA, Gallichotte EN, Ruf IK, Hindson BJ, Vessella RL, Tewari M. Absolute quantification by droplet digital PCR versus analog real-time PCR. Nat Methods. 2013;10:1003–5. https://doi.org/10.1038/nmeth.2633.CrossRefPubMedPubMedCentral

36.

Cieply B, Carstens RP. Functional roles of alternative splicing factors in human disease: functional roles of alternative splicing factors in human disease. Wiley Interdiscip Rev RNA. 2015;6:311–26. https://doi.org/10.1002/wrna.1276.CrossRefPubMedPubMedCentral

37.

Grisendi S, Mecucci C, Falini B, Pandolfi PP. Nucleophosmin and cancer. Nat Rev Cancer. 2006;6:493–505. https://doi.org/10.1038/nrc1885.CrossRefPubMed

38.

Ruggero D, Pandolfi PP. Does the ribosome translate cancer? Nat Rev Cancer. 2003;3:179–92. https://doi.org/10.1038/nrc1015.CrossRefPubMed

39.

Xu D-H, Liu F, Li X, Chen X-F, Jing G-J, Wu F-Y, Shi S-L, Li Q-F. Regulatory role of nucleophosmin during the differentiation of human liver cancer cells. Int J Oncol. 2014;45:264–72. https://doi.org/10.3892/ijo.2014.2407.CrossRefPubMed

40.

Duangmano S, Sae-Lim P, Suksamrarn A, Domann FE, Patmasiriwat P. Cucurbitacin B inhibits human breast cancer cell proliferation through disruption of microtubule polymerization and nucleophosmin/B23 translocation. BMC Complement Altern Med. 2012;12:185. https://doi.org/10.1186/1472-6882-12-185.CrossRefPubMedPubMedCentral

41.

Liu J, Huang B, Xiao Y, Xiong H-M, Li J, Feng D-Q, Chen X-M, Zhang H-B, Wang X-Z. Aberrant expression of splicing factors in newly diagnosed acute myeloid leukemia. Onkologie. 2012;35:335–40. https://doi.org/10.1159/000338941.CrossRefPubMed

42.

Federici L, Falini B. Nucleophosmin mutations in acute myeloid leukemia: a tale of protein unfolding and mislocalization. Protein Sci. 2013;22:545–56. https://doi.org/10.1002/pro.2240.CrossRefPubMedPubMedCentral

43.

Suzuki T, Kiyoi H, Ozeki K, Tomita A, Yamaji S, Suzuki R, Kodera Y, Miyawaki S, Asou N, Kuriyama K, Yagasaki F, Shimazaki C, Akiyama H, et al. Clinical characteristics and prognostic implications of NPM1 mutations in acute myeloid leukemia. Blood. 2005;106:2854–61. https://doi.org/10.1182/blood-2005-04-1733.CrossRefPubMed

44.

Léotoing L, Meunier L, Manin M, Mauduit C, Decaussin M, Verrijdt G, Claessens F, Benahmed M, Veyssière G, Morel L, Beaudoin C. Influence of nucleophosmin/B23 on DNA binding and transcriptional activity of the androgen receptor in prostate cancer cell. Oncogene. 2008;27:2858–67. https://doi.org/10.1038/sj.onc.1210942.CrossRefPubMed

45.

Wang H, Yuan G, Zhao B, Zhao Y, Qiu Y. High expression of B23 is associated with tumorigenesis and poor prognosis in bladder urothelial carcinoma. Mol Med Rep. 2017;15:743–9. https://doi.org/10.3892/mmr.2016.6033.CrossRefPubMed

46.

Zhou F, Chen E, You D, Song Y, Sun Z, Yue L. Both high expression of nucleophosmin/B23 and CRM1 predicts poorer prognosis in human gastric cancer. APMIS Acta Pathol Microbiol Immunol Scand. 2016;124:1046–53. https://doi.org/10.1111/apm.12604.CrossRef

47.

Chen J, Sun J, Yang L, Yan Y, Shi W, Shi J, Huang Q, Chen J, Lan Q. Upregulation of B23 promotes tumor cell proliferation and predicts poor prognosis in glioma. Biochem Biophys Res Commun. 2015;466:124–30. https://doi.org/10.1016/j.bbrc.2015.08.118.CrossRefPubMed

48.

Boissel N, Renneville A, Biggio V, Philippe N, Thomas X, Cayuela J-M, Terre C, Tigaud I, Castaigne S, Raffoux E, De Botton S, Fenaux P, Dombret H, et al. Prevalence, clinical profile, and prognosis of NPM mutations in AML with normal karyotype. Blood. 2005;106:3618–20. https://doi.org/10.1182/blood-2005-05-2174.CrossRefPubMed

49.

Marcucci G, Maharry K, Wu Y-Z, Radmacher MD, Mrózek K, Margeson D, Holland KB, Whitman SP, Becker H, Schwind S, Metzeler KH, Powell BL, Carter TH, et al. IDH1 and IDH2 gene mutations identify novel molecular subsets within de novo cytogenetically normal acute myeloid leukemia: a Cancer and Leukemia Group B study. J Clin Oncol. 2010;28:2348–55. https://doi.org/10.1200/JCO.2009.27.3730.CrossRefPubMedPubMedCentral

Title: NPM1 alternative transcripts are upregulated in acute myeloid and lymphoblastic leukemia and their expression level affects patient outcome
Authors: Luiza Handschuh
Pawel Wojciechowski
Maciej Kazmierczak
Malgorzata Marcinkowska-Swojak
Magdalena Luczak
Krzysztof Lewandowski
Mieczyslaw Komarnicki
Jacek Blazewicz
Marek Figlerowicz
Piotr Kozlowski
Publication date: 01-12-2018
Publisher: BioMed Central
Published in: Journal of Translational Medicine / Issue 1/2018
Electronic ISSN: 1479-5876
DOI: https://doi.org/10.1186/s12967-018-1608-2

Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Pediatric Cardiology Video

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

Cardiopulmonary Comorbidity Video

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Valvular Heart Disease Video

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discuss last year's major advances in heart failure and cardiomyopathies.

ACC 2024 Congress Coverage

Heart Failure Video

Year in Review: Heart failure and cardiomyopathies

Watch now

Watch this official video from ACC.24. Dr. Biykem Bozkurt discuss last year's major advances in heart failure and cardiomyopathies.

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

16-04-2024 Type 2 Diabetes News

Incentivised to exercise

11-04-2024 Lifestyle Modifications News

New intervention for STEMI-related cardiogenic shock

10-04-2024 Myocardial Infarction News

» View more highlights on the ACC 2024 congress hub page

Image Credits

STEP AAG HeroTeaserCollection image/© Tarek / Generated with AI / Stock.adobe.com, ACC 2024 Pediatric and Congenital Heart Disease: Year in Review/© 2024 American College of Cardiology, ACC 2024 Pulmonary Vascular Disease: Year in Review/© 2024 American College of Cardiology, ACC 2024 Valvular Heart Disease: Year in Review/© 2024 American College of Cardiology, ACC 2024 HF and Cardiomyopathies: Year in Review/© 2024 American College of Cardiology, Woman out walking/© Seventyfour / stock.adobe.com (symbolic image with model), Woman checking smartwatch /© saltdium / stock.adobe.com (symbolic image with model), Cardiac catheterization/© Damian / stock.adobe.com

At a glance: The STEP trials

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2018

Clinical feasibility of umbilical cord tissue-derived mesenchymal stem cells in the treatment of multiple sclerosis

Establishment and validation of a predictive nomogram model for non-small cell lung cancer patients with chronic hepatitis B viral infection

Study on the progression types of cancer in patients with breast cancer undergoing eribulin chemotherapy and tumor microenvironment

Detecting germline BAP1 mutations in patients with peritoneal mesothelioma: benefits to patient and family members

Repair of articular cartilage defects with intra-articular injection of autologous rabbit synovial fluid-derived mesenchymal stem cells

Immunomodulatory plasticity of mesenchymal stem cells: a potential key to successful solid organ transplantation

Highlights from the ACC 2024 Congress

ACC 2024 Congress Coverage