Deletion of IKK2 in haematopoietic cells of adult mice leads to elevated interleukin-6, neutrophilia and fatal gastrointestinal inflammation.

Publisher: Nature Pub. Group Country of Publication: England NLM ID: 101524092 Publication Model: Electronic Cited Medium: Internet ISSN: 2041-4889 (Electronic) NLM ISO Abbreviation: Cell Death Dis Subsets: MEDLINE

Original Publication: London : Nature Pub. Group

Gastritis/*immunology ; Hematopoiesis/*immunology ; I-kappa B Kinase/*immunology ; Interleukin-6/*immunology ; Stem Cells/*immunology; Animals ; Cell Differentiation ; Female ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; NF-kappa B/immunology ; Stem Cells/cytology

The IκB kinase complex, consisting of IKK1, IKK2 and the regulatory subunit NEMO, is required for NF-κB signalling following the activation of several cell surface receptors, such as members of the Tumour Necrosis Factor Receptor superfamily and the Interleukin-1 Receptor. This is critical for haematopoietic cell proliferation, differentiation, survival and immune responses. To determine the role of IKK in the regulation of haematopoiesis, we used the Rosa26 ^Cre-ERT2 Cre/lox recombination system to achieve targeted, haematopoietic cell-restricted deletion of the genes for IKK1 or IKK2 in vivo. We found that the IKK complex plays a critical role in haematopoietic cell development and function. Deletion of IKK2, but not loss of IKK1, in haematopoietic cells led to an expansion of CD11b/Gr-1-positive myeloid cells (neutrophilia), severe anaemia and thrombocytosis, with reduced numbers of long-term haematopoietic stem cells (LT-HSCs), short-term haematopoietic stem cells (ST-HSCs) and multipotential progenitor cells (MPPs), increased circulating interleukin-6 (IL-6) and severe gastrointestinal inflammation. These findings identify distinct functions for the two IKK catalytic subunits, IKK1 and IKK2, in the haematopoietic system.

Kindler, V. et al. Stimulation of hematopoiesis in vivo by recombinant bacterial murine interleukin 3. Proc. Natl. Acad. Sci. USA 83, 1001–1005 (1986). (PMID: 308188710.1073/pnas.83.4.1001)
Saeland, S. et al. Effects of recombinant human interleukin-3 on CD34-enriched normal hematopoietic progenitors and on myeloblastic leukemia cells. Blood 72, 1580–1588 (1988). (PMID: 246015610.1182/blood.V72.5.1580.1580)
Brumatti, G., Salmanidis, M. & Ekert, P. G. Crossing paths: interactions between the cell death machinery and growth factor survival signals. Cell. Mol. life Sci. 67, 1619–1630 (2010). (PMID: 2015783810.1007/s00018-010-0288-8)
Chaturvedi, P., Reddy, M. V. & Reddy, E. P. Src kinases and not JAKs activate STATs during IL-3 induced myeloid cell proliferation. Oncogene 16, 1749–1758 (1998). (PMID: 958202310.1038/sj.onc.1201972)
Hercus, T. R. et al. The granulocyte-macrophage colony-stimulating factor receptor: linking its structure to cell signaling and its role in disease. Blood 114, 1289–1298 (2009). (PMID: 19436055272741610.1182/blood-2008-12-164004)
Hansen, G. et al. The structure of the GM-CSF receptor complex reveals a distinct mode of cytokine receptor activation. Cell 134, 496–507 (2008). (PMID: 1869247210.1016/j.cell.2008.05.053)
Rothwarf, D. M., Zandi, E., Natoli, G. & Karin, M. IKK-gamma is an essential regulatory subunit of the IkappaB kinase complex. Nature 395, 297–300 (1998). (PMID: 10.1038/26261)
Yamaoka, S. et al. Complementation cloning of NEMO, a component of the IkappaB kinase complex essential for NF-kappaB activation. Cell 93, 1231–1240 (1998). (PMID: 965715510.1016/S0092-8674(00)81466-X)
DiDonato, J. A., Hayakawa, M., Rothwarf, D. M., Zandi, E. & Karin, M. A cytokine-responsive IkappaB kinase that activates the transcription factor NF-kappaB. Nature 388, 548–554 (1997). (PMID: 925218610.1038/41493)
Mercurio, F. et al. IKK-1 and IKK-2: cytokine-activated IkappaB kinases essential for NF-kappaB activation. Science 278, 860–866 (1997). (PMID: 934648410.1126/science.278.5339.860)
Woronicz, J. D., Gao, X., Cao, Z., Rothe, M. & Goeddel, D. V. IkappaB kinase-beta: NF-kappaB activation and complex formation with IkappaB kinase-alpha and NIK. Science 278, 866–869 (1997). (PMID: 934648510.1126/science.278.5339.866)
Zandi, E., Rothwarf, D. M., Delhase, M., Hayakawa, M. & Karin, M. The IkappaB kinase complex (IKK) contains two kinase subunits, IKKalpha and IKKbeta, necessary for IkappaB phosphorylation and NF-kappaB activation. Cell 91, 243–252 (1997). (PMID: 934624110.1016/S0092-8674(00)80406-7)
Zandi, E., Chen, Y. & Karin, M. Direct phosphorylation of IkappaB by IKKalpha and IKKbeta: discrimination between free and NF-kappaB-bound substrate. Science 281, 1360–1363 (1998). (PMID: 972110310.1126/science.281.5381.1360)
Regnier, C. H. et al. Identification and characterization of an IkappaB kinase. Cell 90, 373–383 (1997). (PMID: 924431010.1016/S0092-8674(00)80344-X)
Perkins, N. D. Integrating cell-signalling pathways with NF-kappaB and IKK function. Nat. Rev. Mol. Cell Biol. 8, 49–62 (2007). (PMID: 1718336010.1038/nrm2083)
Oeckinghaus, A., Hayden, M. S. & Ghosh, S. Crosstalk in NF-kappaB signaling pathways. Nat. Immunol. 12, 695–708 (2011). (PMID: 2177227810.1038/ni.2065)
Gaur, U. & Aggarwal, B. B. Regulation of proliferation, survival and apoptosis by members of the TNF superfamily. Biochem. Pharmacol. 66, 1403–1408 (2003). (PMID: 1455521410.1016/S0006-2952(03)00490-8)
Aggarwal, B. B., Gupta, S. C. & Kim, J. H. Historical perspectives on tumor necrosis factor and its superfamily: 25 years later, a golden journey. Blood 119, 651–665 (2012). (PMID: 22053109326519610.1182/blood-2011-04-325225)
Dejardin, E. et al. The lymphotoxin-beta receptor induces different patterns of gene expression via two NF-kappaB pathways. Immunity 17, 525–535 (2002). (PMID: 1238774510.1016/S1074-7613(02)00423-5)
Claudio, E., Brown, K., Park, S., Wang, H. & Siebenlist, U. BAFF-induced NEMO-independent processing of NF-kappa B2 in maturing B cells. Nat. Immunol. 3, 958–965 (2002). (PMID: 1235296910.1038/ni842)
Coope, H. J. et al. CD40 regulates the processing of NF-kappaB2 p100 to p52. EMBO J. 21, 5375–5385 (2002). (PMID: 1237473812907410.1093/emboj/cdf542)
Kayagaki, N. et al. BAFF/BLyS receptor 3 binds the B cell survival factor BAFF ligand through a discrete surface loop and promotes processing of NF-kappaB2. Immunity 17, 515–524 (2002). (PMID: 1238774410.1016/S1074-7613(02)00425-9)
Mercurio, F. & Manning, A. M. Multiple signals converging on NF-kappaB. Curr. Opin. Cell Biol. 11, 226–232 (1999). (PMID: 1020915710.1016/S0955-0674(99)80030-1)
Senftleben, U. et al. Activation by IKKalpha of a second, evolutionary conserved, NF-kappa B signaling pathway. Science 293, 1495–1499 (2001). (PMID: 1152098910.1126/science.1062677)
Kaisho, T. et al. IkappaB kinase alpha is essential for mature B cell development and function. J. Exp. Med. 193, 417–426 (2001). (PMID: 11181694219590010.1084/jem.193.4.417)
Li, Z. W. et al. The IKKbeta subunit of IkappaB kinase (IKK) is essential for nuclear factor kappaB activation and prevention of apoptosis. J. Exp. Med. 189, 1839–1845 (1999). (PMID: 10359587219308210.1084/jem.189.11.1839)
Tanaka, M. et al. Embryonic lethality, liver degeneration, and impaired NF-kappa B activation in IKK-beta-deficient mice. Immunity 10, 421–429 (1999). (PMID: 1022918510.1016/S1074-7613(00)80042-4)
Makris, C. et al. Female mice heterozygous for IKK gamma/NEMO deficiencies develop a dermatopathy similar to the human X-linked disorder incontinentia pigmenti. Mol. Cell 5, 969–979 (2000). (PMID: 1091199110.1016/S1097-2765(00)80262-2)
Rudolph, D. et al. Severe liver degeneration and lack of NF-kappaB activation in NEMO/IKKgamma-deficient mice. Genes Dev. 14, 854–862 (2000). (PMID: 1076674131649310.1101/gad.14.7.854)
Schmidt-Supprian, M. et al. NEMO/IKK gamma-deficient mice model incontinentia pigmenti. Mol. Cell 5, 981–992 (2000). (PMID: 1091199210.1016/S1097-2765(00)80263-4)
Pasparakis, M., Schmidt-Supprian, M. & Rajewsky, K. IkappaB kinase signaling is essential for maintenance of mature B cells. J. Exp. Med. 196, 743–752 (2002). (PMID: 12235208219405910.1084/jem.20020907)
Li, Z. W., Omori, S. A., Labuda, T., Karin, M. & Rickert, R. C. IKK beta is required for peripheral B cell survival and proliferation. J. Immunol. 170, 4630–4637 (2003). (PMID: 1270734110.4049/jimmunol.170.9.4630)
Schmidt-Supprian, M. et al. Mature T cells depend on signaling through the IKK complex. Immunity 19, 377–389 (2003). (PMID: 1449911310.1016/S1074-7613(03)00237-1)
Schmidt-Supprian, M. et al. Differential dependence of CD4+CD25+ regulatory and natural killer-like T cells on signals leading to NF-kappaB activation. Proc. Natl. Acad. Sci. USA 101, 4566–4571 (2004). (PMID: 1507075810.1073/pnas.0400885101)
Zhang, J., Li, L., Baldwin, A. S. Jr, Friedman, A. D. & Paz-Priel, I. Loss of IKKbeta but not NF-kappaB p65 skews differentiation towards myeloid over erythroid commitment and increases myeloid progenitor self-renewal and functional long-term hematopoietic stem cells. PloS ONE 10, e0130441 (2015). (PMID: 26102347447797810.1371/journal.pone.0130441)
Hsu, L. C. et al. The protein kinase PKR is required for macrophage apoptosis after activation of Toll-like receptor 4. Nature 428, 341–345 (2004). (PMID: 1502920010.1038/nature02405)
Park, J. M. et al. Signaling pathways and genes that inhibit pathogen-induced macrophage apoptosis-CREB and NF-kappaB as key regulators. Immunity 23, 319–329 (2005). (PMID: 1616950410.1016/j.immuni.2005.08.010)
Kanters, E. et al. Inhibition of NF-kappaB activation in macrophages increases atherosclerosis in LDL receptor-deficient mice. J. Clin. Investig. 112, 1176–1185 (2003). (PMID: 1456170210.1172/JCI200318580)
Nai, A. et al. Tamoxifen erythroid toxicity revealed by studying the role of nuclear receptor co-activator 4 in erythropoiesis. Haematologica 104, e383–e384 (2019). (PMID: 31366467666916010.3324/haematol.2019.224857)
Greten, F. R. et al. NF-kappaB is a negative regulator of IL-1beta secretion as revealed by genetic and pharmacological inhibition of IKKbeta. Cell 130, 918–931 (2007). (PMID: 17803913213498610.1016/j.cell.2007.07.009)
Mankan, A. K. et al. TNF-alpha-dependent loss of IKKbeta-deficient myeloid progenitors triggers a cytokine loop culminating in granulocytosis. Proc. Natl Acad. Sci. USA 108, 6567–6572 (2011). (PMID: 2146432010.1073/pnas.1018331108)
Hsu, L. C. et al. IL-1beta-driven neutrophilia preserves antibacterial defense in the absence of the kinase IKKbeta. Nat. Immunol. 12, 144–150 (2011). (PMID: 2117002710.1038/ni.1976)
Brill, J. R. & Baumgardner, D. J. Normocytic anemia. Am. Fam. Phys. 62, 2255–2264 (2000).
Pietras, E. M. et al. Functionally distinct subsets of lineage-biased multipotent progenitors control blood production in normal and regenerative conditions. Cell Stem Cell 17, 35–46 (2015). (PMID: 26095048454215010.1016/j.stem.2015.05.003)
Velasco-Hernandez, T., Sawen, P., Bryder, D. & Cammenga, J. Potential pitfalls of the Mx1-Cre system: implications for experimental modeling of normal and malignant hematopoiesis. Stem Cell Rep. 7, 11–18 (2016). (PMID: 10.1016/j.stemcr.2016.06.002)
Kishimoto, T. The biology of interleukin-6. Blood 74, 1–10 (1989). (PMID: 247379110.1182/blood.V74.1.1.1)
Tecchio, C., Micheletti, A. & Cassatella, M. A. Neutrophil-derived cytokines: facts beyond expression. Front. Immunol. 5, 508 (2014). (PMID: 25374568420463710.3389/fimmu.2014.00508)
Kishimoto, T. IL-6: from its discovery to clinical applications. Int. Immunol. 22, 347–352 (2010). (PMID: 2041025810.1093/intimm/dxq030)
Libermann, T. A. & Baltimore, D. Activation of interleukin-6 gene expression through the NF-kappa B transcription factor. Mol. Cell. Biol. 10, 2327–2334 (1990). (PMID: 2183031360580)
Matsusaka, T. et al. Transcription factors NF-IL6 and NF-kappa B synergistically activate transcription of the inflammatory cytokines, interleukin 6 and interleukin 8. Proc. Natl. Acad. Sci. USA 90, 10193–10197 (1993). (PMID: 823427610.1073/pnas.90.21.10193)
Tanaka, T., Narazaki, M. & Kishimoto, T. IL-6 in inflammation, immunity, and disease. Cold Spring Harb. Perspect. Biol. 6, a016295 (2014). (PMID: 25190079417600710.1101/cshperspect.a016295)
Neurath, M. F. Cytokines in inflammatory bowel disease. Nat. Rev. Immunol. 14, 329–342 (2014). (PMID: 2475195610.1038/nri3661)
Strober, W., Fuss, I. J. & Blumberg, R. S. The immunology of mucosal models of inflammation. Annu Rev. Immunol. 20, 495–549 (2002). (PMID: 1186161110.1146/annurev.immunol.20.100301.064816)
Neurath, M. F. et al. Mucosal Iimmunology (Fourth Edition): Intestinal inflammation and cancer of the gastrointestinal tract Ch. 91, 1761–1775 (Academic Press, Boston, 2015).
De Smaele, E. et al. Induction of gadd45beta by NF-kappaB downregulates pro-apoptotic JNK signalling. Nature 414, 308–313 (2001). (PMID: 1171353010.1038/35104560)
Tang, G. et al. Inhibition of JNK activation through NF-kappaB target genes. Nature 414, 313–317 (2001). (PMID: 1171353110.1038/35104568)
Maeda, S. et al. IKKβ is required for prevention of apoptosis mediated by cell-bound but not by circulating TNFα. Immunity 19, 725–737 (2003). (PMID: 1461485910.1016/S1074-7613(03)00301-7)
Sakon, S. et al. NF-kappaB inhibits TNF-induced accumulation of ROS that mediate prolonged MAPK activation and necrotic cell death. EMBO J. 22, 3898–3909 (2003). (PMID: 1288142416905210.1093/emboj/cdg379)
Kamata, H. et al. Reactive oxygen species promote TNFalpha-induced death and sustained JNK activation by inhibiting MAP kinase phosphatases. Cell 120, 649–661 (2005). (PMID: 1576652810.1016/j.cell.2004.12.041)
Pasparakis, M., Luedde, T. & Schmidt-Supprian, M. Dissection of the NF-kappaB signalling cascade in transgenic and knockout mice. Cell Death Differ. 13, 861–872 (2006). (PMID: 1647022310.1038/sj.cdd.4401870)
Liu, H., Lo, C. R. & Czaja, M. J. NF-kappaB inhibition sensitizes hepatocytes to TNF-induced apoptosis through a sustained activation of JNK and c-Jun. Hepatology 35, 772–778 (2002). (PMID: 1191502210.1053/jhep.2002.32534)
Reuther-Madrid, J. Y. et al. The p65/RelA subunit of NF-kappaB suppresses the sustained, antiapoptotic activity of Jun kinase induced by tumor necrosis factor. Mol. Cell. Biol. 22, 8175–8183 (2002). (PMID: 1241772113407510.1128/MCB.22.23.8175-8183.2002)
Chang, L. & Karin, M. Mammalian MAP kinase signalling cascades. Nature 410, 37–40 (2001). (PMID: 1124203410.1038/35065000)
Mitsuyama, K. et al. Pro-inflammatory signaling by Jun-N-terminal kinase in inflammatory bowel disease. Int. J. Mol. Med. 17, 449–455 (2006). (PMID: 16465391)
Roy, P. K., Rashid, F., Bragg, J. & Ibdah, J. A. Role of the JNK signal transduction pathway in inflammatory bowel disease. World J. Gastroenterol. 14, 200–202 (2008). (PMID: 18186555267511410.3748/wjg.14.200)
Waetzig, G. H., Seegert, D., Rosenstiel, P., Nikolaus, S. & Schreiber, S. p38 mitogen-activated protein kinase is activated and linked to TNF-alpha signaling in inflammatory bowel disease. J. Immunol. 168, 5342–5351 (2002). (PMID: 1199449310.4049/jimmunol.168.10.5342)
Assi, K., Pillai, R., Gomez-Munoz, A., Owen, D. & Salh, B. The specific JNK inhibitor SP600125 targets tumour necrosis factor-alpha production and epithelial cell apoptosis in acute murine colitis. Immunology 118, 112–121 (2006). (PMID: 16630028178226810.1111/j.1365-2567.2006.02349.x)
Davis, R. J. Signal transduction by the JNK group of MAP kinases. Cell 103, 239–252 (2000). (PMID: 1105789710.1016/S0092-8674(00)00116-1)
Johnson, G. L. & Nakamura, K. The c-jun kinase/stress-activated pathway: regulation, function and role in human disease. Biochim. Biophys. Acta 1773, 1341–1348 (2007). (PMID: 17306896199555910.1016/j.bbamcr.2006.12.009)
Ishibashi, T. et al. Interleukin-6 is a potent thrombopoietic factor in vivo in mice. Blood 74, 1241–1244 (1989). (PMID: 278846410.1182/blood.V74.4.1241.1241)
Atreya, R. et al. Blockade of interleukin 6 trans signaling suppresses T-cell resistance against apoptosis in chronic intestinal inflammation: evidence in crohn disease and experimental colitis in vivo. Nat. Med. 6, 583–588 (2000). (PMID: 1080271710.1038/75068)
Jones, S. A. Directing transition from innate to acquired immunity: defining a role for IL-6. J. Immunol. 175, 3463–3468 (2005). (PMID: 1614808710.4049/jimmunol.175.6.3463)
Curnow, S. J. et al. Inhibition of T cell apoptosis in the aqueous humor of patients with uveitis by IL-6/soluble IL-6 receptor trans-signaling. J. Immunol. 173, 5290–5297 (2004). (PMID: 1547007510.4049/jimmunol.173.8.5290)
Gareus, R. et al. Normal epidermal differentiation but impaired skin-barrier formation upon keratinocyte-restricted IKK1 ablation. Nat. Cell Biol. 9, 461–469 (2007). (PMID: 1735163910.1038/ncb1560)
Pasparakis, M. et al. TNF-mediated inflammatory skin disease in mice with epidermis-specific deletion of IKK2. Nature 417, 861–866 (2002). (PMID: 1207535510.1038/nature00820)

0 (Interleukin-6)
0 (NF-kappa B)
0 (interleukin-6, mouse)
EC 2.7.11.10 (Chuk protein, mouse)
EC 2.7.11.10 (I-kappa B Kinase)
EC 2.7.11.10 (Ikbkb protein, mouse)

Date Created: 20210108 Date Completed: 20210913 Latest Revision: 20230127

20230128

PMC7791118

10.1038/s41419-020-03298-9

33414459