Hematopoiesis, the formation of blood cells from hematopoietic stem cells (HSC), is a highly regulated process. Since the discovery of microRNAs (miRNAs), several studies have shown their significant role in the regulation of the hematopoietic system. Impaired expression of miRNAs leads to disrupted cellular pathways and in particular causes loss of hematopoietic ability. Here, we report a previously unrecognized function of miR-143 in granulopoiesis. Hematopoietic cells undergoing granulocytic differentiation exhibited increased miR-143 expression. Overexpression or ablation of miR-143 expression resulted in accelerated granulocytic differentiation or block of differentiation, respectively. The absence of miR-143 in mice resulted in a reduced number of mature granulocytes in blood and bone marrow. Additionally, we observed an association of high miR-143 expression levels with a higher probability of survival in two different cohorts of patients with acute myeloid leukemia (AML). Overexpression of miR-143 in AML cells impaired cell growth, partially induced differentiation, and caused apoptosis. Argonaute2-RNA-Immunoprecipitation assay revealed ERK5, a member of the MAPK-family, as a target of miR-143 in myeloid cells. Further, we observed an inverse correlation of miR-143 and ERK5 in primary AML patient samples, and in CD34+ HSPCs undergoing granulocytic differentiation and we confirmed functional relevance of ERK5 in myeloid cells. In conclusion, our data describe miR-143 as a relevant factor in granulocyte differentiation, whose expression may be useful as a prognostic and therapeutic factor in AML therapy.

Department of Internal Medicine 5 University of Heidelberg Heidelberg Germany

Division of Hematology and Oncology Leipzig University Hospital Leipzig Germany

Institute of Molecular Genetics of the ASCR v v i Laboratory of Haematooncology Prague Czech Republic

Zobrazit více v PubMed

Bartel DP. MicroRNAs: target recognition and regulatory functions. Cell. 2009;136:215–233. doi: 10.1016/j.cell.2009.01.002. PubMed DOI PMC

Lee RC, Feinbaum RL, Ambros V. The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell. 1993;75:843–854. doi: 10.1016/0092-8674(93)90529-Y. PubMed DOI

Brennecke J, Cohen SM. Towards a complete description of the microRNA complement of animal genomes. Genome Biol. 2003;4:228. doi: 10.1186/gb-2003-4-9-228. PubMed DOI PMC

Hatfield SD, et al. Stem cell division is regulated by the microRNA pathway. Nature. 2005;435:974–978. doi: 10.1038/nature03816. PubMed DOI

Guo S, et al. MicroRNA miR-125a controls hematopoietic stem cell number. Proc. Natl. Acad. Sci. USA. 2010;107:14229–14234. doi: 10.1073/pnas.0913574107. PubMed DOI PMC

Fazi F, et al. A minicircuitry comprised of microRNA-223 and transcription factors NFI-A and C/EBPalpha regulates human granulopoiesis. Cell. 2005;123:819–831. doi: 10.1016/j.cell.2005.09.023. PubMed DOI

Bräuer-Hartmann D, et al. PML/RARα-regulated miR-181a/b cluster targets the tumor suppressor RASSF1A in acute promyelocytic leukemia. Cancer Res. 2015;75:3411–3424. doi: 10.1158/0008-5472.CAN-14-3521. PubMed DOI PMC

Gerloff D, et al. NF-κB/STAT5/miR-155 network targets PU.1 in FLT3-ITD-driven acute myeloid leukemia. Leukemia. 2015;29:535–547. doi: 10.1038/leu.2014.231. PubMed DOI PMC

Wurm AA, et al. Disruption of the C/EBPα—miR-182 balance impairs granulocytic differentiation. Nat. Commun. 2017;8:46. doi: 10.1038/s41467-017-00032-6. PubMed DOI PMC

Katzerke C, et al. Transcription factor C/EBPα-induced microRNA-30c inactivates Notch1 during granulopoiesis and is downregulated in acute myeloid leukemia. Blood. 2013;122:2433–2442. doi: 10.1182/blood-2012-12-472183. PubMed DOI PMC

Pulikkan JA, et al. C/EBPα regulated microRNA-34a targets E2F3 during granulopoiesis and is down-regulated in AML with CEBPA mutations. Blood. 2010;116:5638–5649. doi: 10.1182/blood-2010-04-281600. PubMed DOI PMC

Stamato MA, et al. Inhibition of EZH2 triggers the tumor suppressive miR-29b network in multiple myeloma. Oncotarget. 2017;8:106527–106537. doi: 10.18632/oncotarget.22507. PubMed DOI PMC

Ferrara F, Schiffer CA. Acute myeloid leukaemia in adults. Lancet (Lond., Engl.) 2013;381:484–495. doi: 10.1016/S0140-6736(12)61727-9. PubMed DOI

Marcucci G, Haferlach T, Döhner H. Molecular genetics of adult acute myeloid leukemia: prognostic and therapeutic implications. J. Clin. Oncol. 2011;29:475–486. doi: 10.1200/JCO.2010.30.2554. PubMed DOI

Marcucci G, Mrózek K, Radmacher MD, Garzon R, Bloomfield CD. The prognostic and functional role of microRNAs in acute myeloid leukemia. Blood. 2011;117:1121–1129. doi: 10.1182/blood-2010-09-191312. PubMed DOI PMC

Chen X, et al. Clinical value of integrated-signature miRNAs in colorectal cancer: miRNA expression profiling analysis and experimental validation. Oncotarget. 2015;6:37544–37556. PubMed PMC

Votavova H, et al. Differential expression of microRNAs in CD34 + cells of 5q-syndrome. J. Hematol. Oncol. 2011;4:1. doi: 10.1186/1756-8722-4-1. PubMed DOI PMC

Iio A, Nakagawa Y, Hirata I, Naoe T, Akao Y. Identification of non-coding RNAs embracing microRNA-143/145 cluster. Mol. Cancer. 2010;9:136. doi: 10.1186/1476-4598-9-136. PubMed DOI PMC

Cordes KR, et al. miR-145 and miR-143 regulate smooth muscle cell fate and plasticity. Nature. 2009;460:705–710. PubMed PMC

Zhang HP, et al. A regulatory circuit involving miR-143 and DNMT3a mediates vascular smooth muscle cell proliferation induced by homocysteine. Mol. Med. Rep. 2016;13:483–490. doi: 10.3892/mmr.2015.4558. PubMed DOI

Jin YP, et al. miR-143-3p targeting of ITGA6 suppresses tumour growth and angiogenesis by downregulating PLGF expression via the PI3K/AKT pathway in gallbladder carcinoma. Cell Death Dis. 2018;9:182. doi: 10.1038/s41419-017-0258-2. PubMed DOI PMC

Guoping M., Ran L., Yanru Q. miR-143 inhibits cell proliferation of gastric cancer cells through targeting GATA6. Oncol Res. 10.3727/096504018X15151515028670 (2018). PubMed PMC

Zhang Q, Feng Y, Liu P, Yang J. MiR-143 inhibits cell proliferation and invasion by targeting DNMT3A in gastric cancer. Tumour Biol. 2017;39:1010428317711312. PubMed

Kent OA, et al. Repression of the miR-143/145 cluster by oncogenic Ras initiates a tumor-promoting feed-forward pathway. Genes Dev. 2010;24:2754–2759. doi: 10.1101/gad.1950610. PubMed DOI PMC

Lei C, et al. miR-143 and miR-145 inhibit gastric cancer cell migration and metastasis by suppressing MYO6. Cell Death Dis. 2017;8:e3101. doi: 10.1038/cddis.2017.493. PubMed DOI PMC

Piatopoulou, D. et al. Clinical utility of miR-143/miR-182 levels in prognosis and risk stratification specificity of BFM-treated childhood acute lymphoblastic leukemia. Ann. Hematol. . 10.1007/s00277-018-3292-y (2018). PubMed

Drew BA, Burow ME, Beckman BS. MEK5/ERK5 pathway: the first fifteen years. Biochim Biophys. Acta–Rev. Cancer. 2012;1825:37–48. doi: 10.1016/j.bbcan.2011.10.002. PubMed DOI PMC

Xia C, Yang Y, Kong F, Kong Q, Shan C. MiR-143-3p inhibits the proliferation, cell migration and invasion of human breast cancer cells by modulating the expression of MAPK7. Biochimie. 2018;147:98–104. doi: 10.1016/j.biochi.2018.01.003. PubMed DOI

Dong X, et al. MiR-143 regulates the proliferation and migration of osteosarcoma cells through targeting MAPK7. Arch. Biochem. Biophys. 2017;630:47–53. doi: 10.1016/j.abb.2017.07.011. PubMed DOI

Clapé C, et al. miR-143 interferes with ERK5 signaling, and abrogates prostate cancer progression in mice. PLoS One. 2009;4:e7542. doi: 10.1371/journal.pone.0007542. PubMed DOI PMC

Williams CAC, et al. Erk5 is a key regulator of naive-primed transition and embryonic stem cell identity. Cell Rep. 2016;16:1820–1828. doi: 10.1016/j.celrep.2016.07.033. PubMed DOI PMC

Angulo-Ibáñez M, et al. Erk5 contributes to maintaining the balance of cellular nucleotide levels and erythropoiesis. Cell Cycle. 2015;14:3864–3876. doi: 10.1080/15384101.2015.1120914. PubMed DOI PMC

Nithianandarajah-Jones GN, Wilm B, Goldring CEP, Müller J, Cross MJ. ERK5: structure, regulation and function. Cell Signal. 2012;24:2187–2196. doi: 10.1016/j.cellsig.2012.07.007. PubMed DOI

Giurisato E, et al. Myeloid ERK5 deficiency suppresses tumor growth by blocking protumor macrophage polarization via STAT3 inhibition. Proc. Natl. Acad. Sci. USA. 2018;115:E2801–E2810. doi: 10.1073/pnas.1707929115. PubMed DOI PMC

Tusa I, et al. ERK5 is activated by oncogenic BRAF and promotes melanoma growth. Oncogene. 2018;37:2601–2614. doi: 10.1038/s41388-018-0164-9. PubMed DOI PMC

Granados-Jaén A, et al. Absence of ERK5/MAPK7 delays tumorigenesis in Atm-/- mice. Oncotarget. 2016;7:74435–74447. doi: 10.18632/oncotarget.12908. PubMed DOI PMC

Simões AES, Rodrigues CMP, Borralho PM. The MEK5/ERK5 signalling pathway in cancer: a promising novel therapeutic target. Drug Discov. Today. 2016;21:1654–1663. doi: 10.1016/j.drudis.2016.06.010. PubMed DOI

Xu Y, Cao C, Gong X, Rong L. Inhibition of ERK5 enhances cytarabine-induced apoptosis in acute myeloid leukemia cells. Int. J. Clin. Exp. Med. 2015;8:6446–6455. PubMed PMC

Levis M, Small D. FLT3: ITDoes matter in leukemia. Leuk. J. Leuk. Soc. Am. Leuk. Res Fund., UK. 2003;17:1738–1752. PubMed

Croce CM, Calin GA. miRNAs, cancer, and stem cell division. Cell. 2005;122:6–7. doi: 10.1016/j.cell.2005.06.036. PubMed DOI

Zhang B, Pan X, Cobb GP, Anderson TA. microRNAs as oncogenes and tumor suppressors. Dev. Biol. 2007;302:1–12. doi: 10.1016/j.ydbio.2006.08.028. PubMed DOI

Chen L, et al. MicroRNA-143 regulates adipogenesis by modulating the MAP2K5-ERK5 signaling. Sci. Rep. 2014;4:3819. doi: 10.1038/srep03819. PubMed DOI PMC

Batliner J, Buehrer E, Fey MF, Tschan MP. Inhibition of the miR-143/145 cluster attenuated neutrophil differentiation of APL cells. Leuk. Res. 2012;36:237–240. doi: 10.1016/j.leukres.2011.10.006. PubMed DOI

Ng EKO, et al. microRNA-143 is downregulated in breast cancer and regulates DNA methyltransferases 3A in breast cancer cells. Tumour Biol. 2014;35:2591–2598. doi: 10.1007/s13277-013-1341-7. PubMed DOI

Zhou P, Chen WG, Li XW. microRNA-143 acts as a tumor suppressor by targeting hexokinase 2 in human prostate cancer. Am. J. Cancer Res. 2015;5:2056–2063. PubMed PMC

Wei J, et al. miR-143 inhibits cell proliferation by targeting autophagy-related 2B in non-small cell lung cancer H1299 cells. Mol. Med. Rep. 2015;11:571–576. doi: 10.3892/mmr.2014.2675. PubMed DOI

Xia H, et al. miR-143 inhibits NSCLC cell growth and metastasis by targeting Limk1. Int. J. Mol. Sci. 2014;15:11973–11983. doi: 10.3390/ijms150711973. PubMed DOI PMC

Zhang Y, et al. microRNA-143 targets MACC1 to inhibit cell invasion and migration in colorectal cancer. Mol. Cancer. 2012;11:23. doi: 10.1186/1476-4598-11-23. PubMed DOI PMC

Donahue RE, et al. Plerixafor (AMD3100) and granulocyte colony-stimulating factor (G-CSF) mobilize different CD34 + cell populations based on global gene and microRNA expression signatures. Blood. 2009;114:2530–2541. doi: 10.1182/blood-2009-04-214403. PubMed DOI PMC

Merkerova M, Vasikova A, Belickova M, Bruchova H. microRNA expression profiles in umbilical cord blood cell lineages. Stem Cells Dev. 2010;19:17–26. doi: 10.1089/scd.2009.0071. PubMed DOI

Jakob P, Landmesser U. Role of microRNAs in stem/progenitor cells and cardiovascular repair. Cardiovasc. Res. 2012;93:614–622. doi: 10.1093/cvr/cvr311. PubMed DOI

Benati M, et al. Role of JAK2 V617F mutation and aberrant expression of microRNA-143 in myeloproliferative neoplasms. Clin. Chem. Lab. Med. 2015;53:1005–1011. doi: 10.1515/cclm-2014-0858. PubMed DOI

Dou L, et al. Methylation-mediated repression of microRNA-143 enhances MLL–AF4 oncogene expression. Oncogene. 2012;31:507–517. doi: 10.1038/onc.2011.248. PubMed DOI

Volinia S, et al. Reprogramming of miRNA networks in cancer and leukemia. Genome Res. 2010;20:589–599. doi: 10.1101/gr.098046.109. PubMed DOI PMC

Shen JZ, Zhang YY, Fu HY, Wu DS, Zhou HR. Overexpression of microRNA-143 inhibits growth and induces apoptosis in human leukemia cells. Oncol. Rep. 2014;31:2035–2042. doi: 10.3892/or.2014.3078. PubMed DOI

Akao Y, Nakagawa Y, Iio A, Naoe T. Role of microRNA-143 in Fas-mediated apoptosis in human T-cell leukemia Jurkat cells. Leuk. Res. 2009;33:1530–1538. doi: 10.1016/j.leukres.2009.04.019. PubMed DOI

Elhamamsy AR, et al. Circulating miR-92a, miR-143 and miR-342 in plasma are novel potential biomarkers for acute myeloid leukemia. Int. J. Mol. Cell Med. 2017;6:77–86. PubMed PMC

Guo H, et al. The regulation of Toll-like receptor 2 by miR-143 suppresses the invasion and migration of a subset of human colorectal carcinoma cells. Mol. Cancer. 2013;12:77. doi: 10.1186/1476-4598-12-77. PubMed DOI PMC

Avgeris M, et al. Uncovering the clinical utility of miR-143, miR-145 and miR-224 for predicting the survival of bladder cancer patients following treatment. Carcinogenesis. 2015;36:528–537. doi: 10.1093/carcin/bgv024. PubMed DOI

Siemens H, Jackstadt R, Kaller M, Hermeking H. Repression of c-Kit by p53 is mediated by miR-34 and is associated with reduced chemoresistance, migration and stemness. Oncotarget. 2013;4:1399–1415. doi: 10.18632/oncotarget.1202. PubMed DOI PMC

Xia L, et al. miR-15b and miR-16 modulate multidrug resistance by targeting BCL2 in human gastric cancer cells. Int. J. Cancer. 2008;123:372–379. doi: 10.1002/ijc.23501. PubMed DOI

Krakowsky RHE, et al. miR-451a abrogates treatment resistance in FLT3-ITD-positive acute myeloid leukemia. Blood Cancer J. 2018;8:36. doi: 10.1038/s41408-018-0070-y. PubMed DOI PMC

Boren T, et al. MicroRNAs and their target messenger RNAs associated with ovarian cancer response to chemotherapy. Gynecol. Oncol. 2009;113:249–255. doi: 10.1016/j.ygyno.2009.01.014. PubMed DOI

Kopczyńska E. Role of microRNAs in the resistance of prostate cancer to docetaxel and paclitaxel. Współczesna Onkol. 2015;6:423–427. doi: 10.5114/wo.2015.56648. PubMed DOI PMC

Xu B, et al. miR-143 decreases prostate cancer cells proliferation and migration and enhances their sensitivity to docetaxel through suppression of KRAS. Mol. Cell. Biochem. 2011;350:207–213. doi: 10.1007/s11010-010-0700-6. PubMed DOI

Wang L, et al. MiR-143 acts as a tumor suppressor by targeting N-RAS and enhances temozolomide-induced apoptosis in glioma. Oncotarget. 2014;5:5416–5427. PubMed PMC

Gomes SE, et al. Convergence of miR-143 overexpression, oxidative stress and cell death in HCT116 human colon cancer cells. PLoS One. 2018;13:e0191607. doi: 10.1371/journal.pone.0191607. PubMed DOI PMC

Kim SC, et al. Constitutive activation of extracellular signal-regulated kinase in human acute leukemias: combined role of activation of MEK, hyperexpression of extracellular signal-regulated kinase, and downregulation of a phosphatase, PAC1. Blood. 1999;93:3893–3899. PubMed

Towatari M, et al. Constitutive activation of mitogen-activated protein kinase pathway in acute leukemia cells. Leukemia. 1997;11:479–484. doi: 10.1038/sj.leu.2400617. PubMed DOI

Perez-Madrigal D, Finegan KG, Paramo B, Tournier C. The extracellular-regulated protein kinase 5 (ERK5) promotes cell proliferation through the down-regulation of inhibitors of cyclin dependent protein kinases (CDKs) Cell. Signal. 2012;24:2360–2368. doi: 10.1016/j.cellsig.2012.08.001. PubMed DOI

Garaude J, et al. ERK5 activates NF-kappaB in leukemic T cells and is essential for their growth in vivo. J. Immunol. 2006;177:7607–7617. doi: 10.4049/jimmunol.177.11.7607. PubMed DOI

Zheng R, Studzinski GP. Optimal AraC-cytotoxicity to aml cells requires erk5 activity. J. Cell. Biochem. 2017;118:1583–1589. doi: 10.1002/jcb.25820. PubMed DOI

Ahmad I, et al. Mir143 expression inversely correlates with nuclear ERK5 immunoreactivity in clinical prostate cancer. Br. J. Cancer. 2013;108:149–154. doi: 10.1038/bjc.2012.510. PubMed DOI PMC

Wang X, Pesakhov S, Harrison JS, Danilenko M, Studzinski GP. ERK5 pathway regulates transcription factors important for monocytic differentiation of human myeloid leukemia cells. J. Cell. Physiol. 2014;229:856–867. doi: 10.1002/jcp.24513. PubMed DOI PMC

Carvajal-Vergara X, et al. Multifunctional role of Erk5 in multiple myeloma. Blood. 2005;105:4492–4499. doi: 10.1182/blood-2004-08-2985. PubMed DOI

Piloto O, et al. Prolonged exposure to FLT3 inhibitors leads to resistance via activation of parallel signaling pathways. Blood. 2007;109:1643–1652. doi: 10.1182/blood-2006-05-023804. PubMed DOI PMC

McCubrey JA, et al. Roles of the Raf/MEK/ERK pathway in cell growth, malignant transformation and drug resistance. Biochim Biophys. Acta–Mol. Cell Res. 2007;1773:1263–1284. doi: 10.1016/j.bbamcr.2006.10.001. PubMed DOI PMC

McCubrey JA, et al. Ras/Raf/MEK/ERK and PI3K/PTEN/Akt/mTOR cascade inhibitors: how mutations can result in therapy resistance and how to overcome resistance. Oncotarget. 2012;3:1068–1111. PubMed PMC

Zjablovskaja P, et al. EVI2B is a C/EBPα target gene required for granulocytic differentiation and functionality of hematopoietic progenitors. Cell Death Differ. 2017;24:705–716. doi: 10.1038/cdd.2017.6. PubMed DOI PMC

Trapnell C, et al. Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and Cufflinks. Nat. Protoc. 2012;7:562–578. doi: 10.1038/nprot.2012.016. PubMed DOI PMC