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Ablation of miR-146b in mice causes hematopoietic malignancy

Mitsumura, Takahiro, Ito, Yoshiaki, Chiba, Tomoki, Matsushima, Takahide, Kurimoto, Ryota, Tanaka, Yoko, Kato, Tomomi, Uchida, Keisuke, Ito, Takashi, Yamamoto, Kouhei, Eishi, Yoshinobu, Kitagawa, Masanobu, Miyazaki, Yasunari, Inase, Naohiko, Asahara, Hiroshi·2018·Blood Advances
DOI 10.1182/bloodadvances.2018017954
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Abstract

Excessive and constitutive activation of nuclear factor-κB (NF-κB) leads to abnormal cell proliferation and differentiation, leading to the development of malignant tumors, including lymphoma. MicroRNA 146a (miR-146a) and miR-146b, both of which carry an identical seed sequence, have been shown to contribute to inflammatory diseases and tumors by suppressing the expression of key molecules required for NF-κB activation. However, the functional and physiological differences between miR-146a and miR-146b in disease onset have not been fully elucidated. In this study, we generated miR-146b–knockout (KO) and miR-146a–KO mice by genome editing and found that both strains developed hematopoietic malignancies such as B-cell lymphoma and acute myeloid leukemia during aging. However, the B-cell lymphomas observed in miR-146a– and miR-146b–KO mice were histologically different in their morphology, and the malignancy rate is lower in miR-146b mice than miR-146a mice. Upon mitogenic stimulation, the expression of miR-146a and miR-146b was increased, but miR-146b expression was lower than that of miR-146a. Using a previously developed screening system for microRNA targets, we observed that miR-146a and miR-146b could target the same mRNAs, including TRAF6, and inhibit subsequent NF-κB activity. Consistent with these findings, both miR-146a– and miR-146b–KO B cells showed a high proliferative capacity. Taken together, sustained NF-κB activation in miR-146b KO mice could lead to the development of hematopoietic malignancy with aging.

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Figure 1 Production of miR-146a and miR-146b KO mice with TALEN. (A) Genetic deletion in miR-146a -and miR-146b -deficient mice generated by TALEN. The genomic sequence is shown at the top, with the target sequences of a pair of TALEN colored in red. Blue lines and boxes indicate pre-miRNA and mature miRNA sequences, respectively. Deleted nucleotides are indicated with dashes. The sizes of the deletions and insertions are shown to the right of the mutated alleles with D . WT indicates alleles without a mutation. (B) RT-PCR analysis of miR-146a and miR-146b in liver tissue derived from mutant mice. The genotype of each animal is indicated at the bottom. The expression level of miR-146a and miR-146b in the WT animal was adjusted to 1.
Figure 1 Production of miR-146a and miR-146b KO mice with TALEN. (A) Genetic deletion in miR-146a -and miR-146b -deficient mice generated by TALEN. The genomic sequence is shown at the top, with the target sequences of a pair of TALEN colored in red. Blue lines and boxes indicate pre-miRNA and mature miRNA sequences, respectively. Deleted nucleotides are indicated with dashes. The sizes of the deletions and insertions are shown to the right of the mutated alleles with D . WT indicates alleles without a mutation. (B) RT-PCR analysis of miR-146a and miR-146b in liver tissue derived from mutant mice. The genotype of each animal is indicated at the bottom. The expression level of miR-146a and miR-146b in the WT animal was adjusted to 1.CC-BY-NC-ND · From the PDF
Figure 2 Histological analysis of spleens and lymph nodes in WT mice and representative malignancies in miR-146a or miR-146b KO mice. (A) Increased rate of tumorigenesis in mice with miR-146a or miR-146b ablation. n 5 11 (WT), 8 (miR-146a KO1), 3 (miR-146a KO2), and 21 (miR-146b KO). (B) Photograph of spleens isolated from WT and miR-146b KO mice at 16 months. (C) miR-146a and miR-146b KO mice developed B-cell lymphomas in the spleen and lymph nodes. Scale bars represent 500 m m (low magnification), 20 m m (high magnification), and 2.5 m m (insets). (D) miR-146b KO mice developed acute leukemia. Scale bars represent 500 m m (low magnification) and 20 m m (high magnification), and 2.5 m m (inset). HE, hematoxylin and eosin; FL, follicular lymphoma.
Figure 2 Histological analysis of spleens and lymph nodes in WT mice and representative malignancies in miR-146a or miR-146b KO mice. (A) Increased rate of tumorigenesis in mice with miR-146a or miR-146b ablation. n 5 11 (WT), 8 (miR-146a KO1), 3 (miR-146a KO2), and 21 (miR-146b KO). (B) Photograph of spleens isolated from WT and miR-146b KO mice at 16 months. (C) miR-146a and miR-146b KO mice developed B-cell lymphomas in the spleen and lymph nodes. Scale bars represent 500 m m (low magnification), 20 m m (high magnification), and 2.5 m m (insets). (D) miR-146b KO mice developed acute leukemia. Scale bars represent 500 m m (low magnification) and 20 m m (high magnification), and 2.5 m m (inset). HE, hematoxylin and eosin; FL, follicular lymphoma.CC-BY-NC-ND · From the PDF
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CC-BY-NC-ND · From the PDF
Figure 4 Suppression of NFk B activity by miR-146a and miR-146b. (A) Luciferase reporter assay for miR-146a -or miR-146b -dependent regulation of NFk B. Luciferase activity was measured in HEK293T cells transfected with a control empty vector, miR-146a, or miR-146b expression vector (with or without 50 ng/mL PMA). Data are presented as the mean 6 SEM. Two independent experiments were performed, and similar results were obtained. 1 , 5 ng; 11 , 75 ng. (B) Luciferase reporter assays in 293FT cells transfected with pcDNA-miR-146a (miR-146a), pcDNA-miR-146b (miR-146b), or an empty vector (Ctrl). MCS, pLuc2-KAP-MCS (multicloning site as an empty reporter). Data are presented as the mean 6 SEM; n 5 3. (C) Western blot analysis of TRAF6 protein expression in nonstimulated WT and miR-146a or miR-146b KO splenic B cells and those stimulated with LPS or anti-CD40 antibody. Two mice per genotype were analyzed; representative results are shown. * P , .05, ** P , .01.
Figure 4 Suppression of NFk B activity by miR-146a and miR-146b. (A) Luciferase reporter assay for miR-146a -or miR-146b -dependent regulation of NFk B. Luciferase activity was measured in HEK293T cells transfected with a control empty vector, miR-146a, or miR-146b expression vector (with or without 50 ng/mL PMA). Data are presented as the mean 6 SEM. Two independent experiments were performed, and similar results were obtained. 1 , 5 ng; 11 , 75 ng. (B) Luciferase reporter assays in 293FT cells transfected with pcDNA-miR-146a (miR-146a), pcDNA-miR-146b (miR-146b), or an empty vector (Ctrl). MCS, pLuc2-KAP-MCS (multicloning site as an empty reporter). Data are presented as the mean 6 SEM; n 5 3. (C) Western blot analysis of TRAF6 protein expression in nonstimulated WT and miR-146a or miR-146b KO splenic B cells and those stimulated with LPS or anti-CD40 antibody. Two mice per genotype were analyzed; representative results are shown. * P , .05, ** P , .01.CC-BY-NC-ND · From the PDF
Figure 5 Proliferation of WT, miR-146a KO, and miR-146b KO B cells in response to LPS, anti-CD40, and anti-IgM antibody. Splenic B cells (CD19 1 ) were incubated for 48 hours in the presence of media only (none) or the indicated mitogens (0.1, 1, or 10 m g/mL LPS; 0.1, 1, or 5 m g/mL anti-CD40; and 10 m g/mL anti-IgM). Proliferation was measured by [ 3 H]-thymidine incorporation. Data are presented as mean 6 SEM. n 5 6. * P , .05, ** P , .01. cpm, counts per minute.
Figure 5 Proliferation of WT, miR-146a KO, and miR-146b KO B cells in response to LPS, anti-CD40, and anti-IgM antibody. Splenic B cells (CD19 1 ) were incubated for 48 hours in the presence of media only (none) or the indicated mitogens (0.1, 1, or 10 m g/mL LPS; 0.1, 1, or 5 m g/mL anti-CD40; and 10 m g/mL anti-IgM). Proliferation was measured by [ 3 H]-thymidine incorporation. Data are presented as mean 6 SEM. n 5 6. * P , .05, ** P , .01. cpm, counts per minute.CC-BY-NC-ND · From the PDF

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Research works printed but not linked: 52

  1. Bartel, 2009, MicroRNAs: target recognition and regulatory functions, Cell, 136, 215, 10.1016/j.cell.2009.01.002 · DOI · Paper page
  2. Boldin, 2012, MicroRNAs, new effectors and regulators of NF-κB, Immunol Rev, 246, 205, 10.1111/j.1600-065X.2011.01089.x · DOI · Paper page
  3. Wallace, 2017, MicroRNAs and acute myeloid leukemia: therapeutic implications and emerging concepts, Blood, 130, 1290, 10.1182/blood-2016-10-697698 · DOI
  4. Braun, 2006, Targeting NF-kappaB in hematologic malignancies, Cell Death Differ, 13, 748, 10.1038/sj.cdd.4401874 · DOI
  5. Shen, 2009, NFkappaB signaling in carcinogenesis and as a potential molecular target for cancer therapy, Apoptosis, 14, 348, 10.1007/s10495-009-0315-0 · DOI
  6. Young, 2013, Targeting pathological B cell receptor signalling in lymphoid malignancies, Nat Rev Drug Discov, 12, 229, 10.1038/nrd3937 · DOI
  7. Compagno, 2009, Mutations of multiple genes cause deregulation of NF-kappaB in diffuse large B-cell lymphoma, Nature, 459, 717, 10.1038/nature07968 · DOI
  8. Kato, 2009, Frequent inactivation of A20 in B-cell lymphomas, Nature, 459, 712, 10.1038/nature07969 · DOI
  9. Lenz, 2008, Oncogenic CARD11 mutations in human diffuse large B cell lymphoma, Science, 319, 1676, 10.1126/science.1153629 · DOI
  10. Ngo, 2011, Oncogenically active MYD88 mutations in human lymphoma, Nature, 470, 115, 10.1038/nature09671 · DOI
  11. Pasqualucci, 2011, Analysis of the coding genome of diffuse large B-cell lymphoma, Nat Genet, 43, 830, 10.1038/ng.892 · DOI
  12. Davis, 2010, Chronic active B-cell-receptor signalling in diffuse large B-cell lymphoma, Nature, 463, 88, 10.1038/nature08638 · DOI
  13. Taganov, 2006, NF-kappaB-dependent induction of microRNA miR-146, an inhibitor targeted to signaling proteins of innate immune responses, Proc Natl Acad Sci USA, 103, 12481, 10.1073/pnas.0605298103 · DOI · Paper page
  14. Lu, 2010, Function of miR-146a in controlling Treg cell-mediated regulation of Th1 responses, Cell, 142, 914, 10.1016/j.cell.2010.08.012 · DOI · Paper page
  15. Zhao, 2011, NF-kappaB dysregulation in microRNA-146a-deficient mice drives the development of myeloid malignancies, Proc Natl Acad Sci USA, 108, 9184, 10.1073/pnas.1105398108 · DOI · Paper page
  16. Yang, 2012, miR-146a controls the resolution of T cell responses in mice, J Exp Med, 209, 1655, 10.1084/jem.20112218 · DOI
  17. Boldin, 2011, miR-146a is a significant brake on autoimmunity, myeloproliferation, and cancer in mice, J Exp Med, 208, 1189, 10.1084/jem.20101823 · DOI · Paper page
  18. Bhaumik, 2008, Expression of microRNA-146 suppresses NF-kappaB activity with reduction of metastatic potential in breast cancer cells, Oncogene, 27, 5643, 10.1038/onc.2008.171 · DOI
  19. Hurst, 2009, Breast cancer metastasis suppressor 1 up-regulates miR-146, which suppresses breast cancer metastasis, Cancer Res, 69, 1279, 10.1158/0008-5472.CAN-08-3559 · DOI
  20. Xia, 2009, microRNA-146b inhibits glioma cell migration and invasion by targeting MMPs, Brain Res, 1269, 158, 10.1016/j.brainres.2009.02.037 · DOI
  21. Cai, 2015, MicroRNA-146b-5p inhibits the growth of gallbladder carcinoma by targeting epidermal growth factor receptor, Mol Med Rep, 12, 1549, 10.3892/mmr.2015.3461 · DOI
  22. Wu, 2014, Low expression of microRNA-146b-5p and microRNA-320d predicts poor outcome of large B-cell lymphoma treated with cyclophosphamide, doxorubicin, vincristine, and prednisone, Hum Pathol, 45, 1664, 10.1016/j.humpath.2014.04.002 · DOI
  23. Burger, 2014, Premalignant PTEN-deficient thymocytes activate microRNAs miR-146a and miR-146b as a cellular defense against malignant transformation, Blood, 123, 4089, 10.1182/blood-2013-11-539411 · DOI
  24. Correia, 2016, MiR-146b negatively regulates migration and delays progression of T-cell acute lymphoblastic leukemia, Sci Rep, 6, 31894, 10.1038/srep31894 · DOI
  25. Xiang, 2014, STAT3 induction of miR-146b forms a feedback loop to inhibit the NF-κB to IL-6 signaling axis and STAT3-driven cancer phenotypes, Sci Signal, 7, ra11, 10.1126/scisignal.2004497 · DOI
  26. Li, 2013, miR-146b-5p inhibits glioma migration and invasion by targeting MMP16, Cancer Lett, 339, 260, 10.1016/j.canlet.2013.06.018 · DOI
  27. Liu, 2015, miR-146b-5p functions as a tumor suppressor by targeting TRAF6 and predicts the prognosis of human gliomas, Oncotarget, 6, 29129, 10.18632/oncotarget.4895 · DOI
  28. Takada, 2013, Targeted gene deletion of miRNAs in mice by TALEN system, PLoS One, 8, e76004, 10.1371/journal.pone.0076004 · DOI
  29. Ito, 2017, Identification of targets of tumor suppressor microRNA-34a using a reporter library system, Proc Natl Acad Sci USA, 114, 3927, 10.1073/pnas.1620019114 · DOI
  30. Kim, 2013, TALEN-based knockout library for human microRNAs, Nat Struct Mol Biol, 20, 1458, 10.1038/nsmb.2701 · DOI
  31. Cheng, 2013, MicroRNA-146 represses endothelial activation by inhibiting pro-inflammatory pathways, EMBO Mol Med, 5, 1017, 10.1002/emmm.201202318 · DOI
  32. Hans, 2004, Confirmation of the molecular classification of diffuse large B-cell lymphoma by immunohistochemistry using a tissue microarray, Blood, 103, 275, 10.1182/blood-2003-05-1545 · DOI
  33. Perry, 2008, Rapid changes in microRNA-146a expression negatively regulate the IL-1beta-induced inflammatory response in human lung alveolar epithelial cells, J Immunol, 180, 5689, 10.4049/jimmunol.180.8.5689 · DOI · Paper page
  34. Hou, 2009, MicroRNA-146a feedback inhibits RIG-I-dependent Type I IFN production in macrophages by targeting TRAF6, IRAK1, and IRAK2, J Immunol, 183, 2150, 10.4049/jimmunol.0900707 · DOI · Paper page
  35. Nahid, 2011, Mechanistic role of microRNA-146a in endotoxin-induced differential cross-regulation of TLR signaling, J Immunol, 186, 1723, 10.4049/jimmunol.1002311 · DOI
  36. Etzrodt, 2012, Regulation of monocyte functional heterogeneity by miR-146a and Relb, Cell Reports, 1, 317, 10.1016/j.celrep.2012.02.009 · DOI
  37. Quinn, 2013, MicroRNA-146a is upregulated by and negatively regulates TLR2 signaling, PLoS One, 8, e62232, 10.1371/journal.pone.0062232 · DOI
  38. Monk, 2010, Regulation of miRNA transcription in macrophages in response to Candida albicans, PLoS One, 5, e13669, 10.1371/journal.pone.0013669 · DOI
  39. Curtale, 2013, Negative regulation of Toll-like receptor 4 signaling by IL-10-dependent microRNA-146b, Proc Natl Acad Sci USA, 110, 11499, 10.1073/pnas.1219852110 · DOI
  40. Park, 2015, MicroRNA-146a and microRNA-146b regulate human dendritic cell apoptosis and cytokine production by targeting TRAF6 and IRAK1 proteins, J Biol Chem, 290, 2831, 10.1074/jbc.M114.591420 · DOI
  41. He, 2016, MicroR-146 blocks the activation of M1 macrophage by targeting signal transducer and activator of transcription 1 in hepatic schistosomiasis, EBioMedicine, 13, 339, 10.1016/j.ebiom.2016.10.024 · DOI
  42. Kanie, 2017, The CEP19-RABL2 GTPase complex binds IFT-B to initiate intraflagellar transport at the ciliary base, Dev Cell, 42, 22, 10.1016/j.devcel.2017.05.016 · DOI
  43. Yi Lo, 2016, RABL2 is required for hepatic fatty acid homeostasis and its dysfunction leads to steatosis and a diabetes-like state, Endocrinology, 157, 4732, 10.1210/en.2016-1487 · DOI
  44. Lo, 2012, RAB-like 2 has an essential role in male fertility, sperm intra-flagellar transport, and tail assembly, PLoS Genet, 8, e1002969, 10.1371/journal.pgen.1002969 · DOI
  45. Xie, 2015, The causative gene in Chanarian Dorfman syndrome regulates lipid droplet homeostasis in C. elegans, PLoS Genet, 11, e1005284, 10.1371/journal.pgen.1005284 · DOI
  46. Perkins, 2012, The diverse and complex roles of NF-κB subunits in cancer, Nat Rev Cancer, 12, 121, 10.1038/nrc3204 · DOI
  47. Gilmore, 2006, Introduction to NF-kappaB: players, pathways, perspectives, Oncogene, 25, 6680, 10.1038/sj.onc.1209954 · DOI
  48. Magilnick, 2017, miR-146a-Traf6 regulatory axis controls autoimmunity and myelopoiesis, but is dispensable for hematopoietic stem cell homeostasis and tumor suppression, Proc Natl Acad Sci USA, 114, E7140, 10.1073/pnas.1706833114 · DOI
  49. Zhou, 2015, Aberrant nuclear factor-kappa B activity in acute myeloid leukemia: from molecular pathogenesis to therapeutic target, Oncotarget, 6, 5490, 10.18632/oncotarget.3545 · DOI
  50. Zhai, 2014, The regulatory roles of microRNA-146b-5p and its target platelet-derived growth factor receptor α (PDGFRA) in erythropoiesis and megakaryocytopoiesis, J Biol Chem, 289, 22600, 10.1074/jbc.M114.547380 · DOI
  51. Cho, 2018, Differential cell-intrinsic regulations of germinal center B and T cells by miR-146a and miR-146b, Nat Commun, 9, 2757, 10.1038/s41467-018-05196-3 · DOI
  52. Starczynowski, 2010, Identification of miR-145 and miR-146a as mediators of the 5q- syndrome phenotype, Nat Med, 16, 49, 10.1038/nm.2054 · DOI · Paper page

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Metadata

Authors
Mitsumura, T., Ito, Y., Chiba, T., Matsushima, T., Kurimoto, R., Tanaka, Y., Kato, T., Uchida, K., Ito, T., Yamamoto, K., Eishi, Y., Kitagawa, M., Miyazaki, Y., Inase, N., & Asahara, H.
Affiliations
  • Department of Systems BioMedicine, Graduate School of Medical and Dental Sciences,
  • Division of Surgical Pathology, Tokyo Medical and Dental University Hospital, Tokyo, Japan;
  • Department of Human Pathology and
  • Department of Comprehensive Pathology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan; and
  • Department of Respiratory Medicine, Graduate School of Medical and Dental Sciences, and
ORCID
Venue
Blood Advances
Publisher
American Society of Hematology
Published
2018-12-07
Volume
2
Issue
23
Pages
3483-3491
ISSN
  • 2473-9529
  • 2473-9537
DOI
10.1182/bloodadvances.2018017954
Cited by
22
References
4
Language
en

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