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Key Event Title
FOXJ1 Protein, Decreased
|Level of Biological Organization|
|multi-ciliated epithelial cell|
Key Event Components
|forkhead box protein J1||decreased|
Key Event Overview
AOPs Including This Key Event
|All life stages||High|
Key Event Description
The epithelium of the respiratory tract has a powerful defense mechanism against air-borne pollutants due to the combined performance of mucus-producing goblet cells and ciliated cells that are covered with microtubule-based projections, the cilia. In response to various irritants and pathogens mucus is secreted by goblet cells, and cilia sweep mucus upward by coordinated beating motions thus clearing the airways from these substances. The ciliated airway epithelial cells are typically covered by hundreds of motile cilia. Cilia formation is initiated and coordinated by a distinct gene expression program, led by the transcription factor forkhead box J1 (FOXJ1) (Brody et al., 2000; Zhou and Roy, 2015). In addition to the respiratory tract, FOXJ1 is expressed also in the ciliated cells of the reproductive and central nervous systems (Blatt et al., 1999; Hackett et al., 1995; Lim et al., 1997).
The multiple motile cilia assembly factors MCIDAS and GMNC converge in positive regulation of FOXJ1 (Arbi et al., 2016; Berta et al., 2016; Stubbs et al., 2012), whereas NOTCH signaling, IL-13-or EGF (epidermal growth factor)-triggered signaling antagonize FOXJ1-driven multiciliogenesis (Gerovac and Fregien, 2016; Gerovac et al., 2014; Gomperts et al., 2007; Shaykhiev et al., 2013). Various other factors are involved in multiple motile cilia assembly, including MYB (acts early in multiciliogenesis downstream of MCIDAS), RFX3 (can act as a co-factor for FOXJ1), ULK4 (modulates the expression of FOXJ1), Wnt signaling, etc. (Choksi et al., 2014; Liu et al., 2016; Schmid et al., 2017; Tan et al., 2013). Most of these factors act upstream or parallel to FOXJ1. FOXJ1 appears to be the major factor in multiciliogenesis, whereby its activity is necessary and also sufficient for programming cells to assemble functional motile cilia (Vij et al., 2012).
FOXJ1 is a master regulator of motile ciliogenesis and is essential to program cells to grow motile cilia (Zhou and Roy, 2015). This key event represents the decrease in the levels or absence of FOXJ1 protein in cells of the respiratory tract. The decrease in FOXJ1 levels inhibits ciliogenesis in multiciliated cells of zebrafish and Xenopus (Stubbs et al., 2008). The knockdown of FOXJ1 results in almost complete absence of cilia in mouse epithelial cells (Brody et al., 2000; Chen J. et al., 1998). On the other hand, the overexpression of FOXJ1 rescues cigarette smoke-mediated suppression of cilia growth in human airway epithelium (Brekman et al., 2014).
How It Is Measured or Detected
FOXJ1 protein levels can be measured by Western blot analysis (Brekman et al., 2014; Didon et al., 2013a; Gomperts et al., 2007; Jacquet et al., 2009; Milara et al., 2012), immunofluorescence (Arbi et al., 2016; Gomperts et al., 2007; Valencia-Gattas et al., 2016) or immunohistochemistry (Abedalthagafi et al., 2016; Danielian et al., 2007; Gao et al., 2015). FOXJ1 protein amounts can be inferred from FOXJ1 mRNA levels that can be measured by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) (Arbi et al., 2016; Brekman et al., 2014; Didon et al., 2013a; Jacquet et al., 2009; Milara et al., 2012; Stubbs et al., 2012), in situ hybridization (Hackett et al., 1995; Stubbs et al., 2012), and Northern blot analysis (Hackett et al., 1995). In addition, FOXJ1 protein activity can be inferred from FOXJ1 target gene expression levels or from reporter gene expression levels (e.g. luciferase assay) of genes harboring FOXJ1 transcription factor binding sites (Brekman et al., 2014; Lim et al., 1997).
Domain of Applicability
FOXJ1 is functionally conserved throughout diverse groups of metazoans including flatworm Schmidtea mediterranea, zebrafish Danio rerio, African clawed frog Xenopus laevis (Stubbs et al., 2008; Vij et al., 2012; Yu et al., 2008). Ectopic expression of FOXJ1 triggers ciliogenesis in zebrafish and frog (Stubbs et al., 2008; Yu et al., 2008). Overexpression of FOXJ1 transcription factor in the neural tube of a chick induces cilia formation (Cruz C. et al., 2010). There are multiple studies of FOXJ1 in mice and in human cells (Boon et al., 2014; Brekman et al., 2014; Brody et al., 2000; Chen et al., 1998; Choksi et al., 2014). Furthermore, the target genes of FOXJ1, for example RFX3, are regulated by FOXJ1 across different species (Alten et al., 2012; Didon et al., 2013a).
FOXJ1 function is important for all life stages from embryo through adulthood (Choksi et al., 2014; Stauber et al., 2017).
FOXJ1 is expressed in the airways of both males and females. In addition to respiratory tract and brain, FOXJ1 is functionally important also in male and female reproductive tissues (Hackett et al., 1995).
Evidence for Perturbation by Stressor
Whole cigarette smoke exposure or treatment with cigarette smoke extract of normal human bronchial epithelial cells significantly lowered FoxJ1 mRNA and protein levels (Milara et al., 2012; Brekman et al., 2014; Valencia-Gattas et al., 2016; Ishikawa and Ito, 2017). Cigarette smoke extract treatment of normal human bronchial epithelial cells also reduced the expression of cilia-related transcription factor genes, including FOXJ1, RFX2, and RFX3, as well as that of cilia motility and structural integrity genes regulated by FOXJ1, including DNAI1, DNAH5, DNAH9, DNAH10, DNAH11, and SPAG6 (Brekman et al., 2014).
Irradiation causes excessive levels of free radicals and associated lipid peroxidation, damage to DNA, proteins, leading to wide-spread cellular damage (Azzam et al., 2012; Koc et al., 2003; Rodrigues-Moreira et al., 2017; Shirazi et al., 2013). Thoracic irradiation reduces FOXJ1 mRNA levels in mouse lungs (Bernard et al., 2012).
Alten, L., Schuster-Gossler, K., Beckers, A., Groos, S., Ulmer, B., Hegermann, J., et al. (2012). Differential regulation of node formation, nodal ciliogenesis and cilia positioning by Noto and Foxj1. Development 139, 1276-1284.
Arbi, M., Pefani, D.E., Kyrousi, C., Lalioti, M.E., Kalogeropoulou, A., Papanastasiou, A.D., et al. (2016). GemC1 controls multiciliogenesis in the airway epithelium. EMBO Rep. 17, 400-413.
Azzam, E.I., Jay-Gerin, J.P. and Pain, D. (2012). Ionizing radiation-induced metabolic oxidative stress and prolonged cell injury. Cancer Lett. 327, 48-60.
Bernard, M.E., Kim, H., Rajagopalan, M.S., Stone, B., Salimi, U., Rwigema, J.C., et al. (2012). Repopulation of the irradiation damaged lung with bone marrow-derived cells. In Vivo. 26, 9-18.
Berta, T., Gabriele, P., Sandra, S.-B., Gabriel, G.-G., A, Y.S., Stephan-Otto, A.C., et al. (2016). GEMC1 is a critical regulator of multiciliated cell differentiation. EMBO J. 35, 942-960.
Blatt, E.N., Yan, X.H., Wuerffel, M.K., Hamilos, D.L. and Brody, S.L. (1999). Forkhead transcription factor HFH-4 expression is temporally related to ciliogenesis. Am. J. Respir. Cell Mol. Biol. 21, 168-176.
Boon, M., Wallmeier, J., Ma, L., Loges, N.T., Jaspers, M., Olbrich, H., et al. (2014). MCIDAS mutations result in a mucociliary clearance disorder with reduced generation of multiple motile cilia. Nat. Commun. 5, 4418.
Brekman, A., Walters, M.S., Tilley, A.E. and Crystal, R.G. (2014). FOXJ1 prevents cilia growth inhibition by cigarette smoke in human airway epithelium in vitro. Am. J. Respir. Cell Mol. Biol. 51, 688-700.
Brody, S.L., Yan, X.H., Wuerffel, M.K., Song, S.K. and Shapiro, S.D. (2000). Ciliogenesis and left-right axis defects in forkhead factor HFH-4-null mice. Am. J. Respir. Cell Mol. Biol. 23, 45-51.
Chen, J., Knowles, H.J., Hebert, J.L. and Hackett, B.P. (1998). Mutation of the mouse hepatocyte nuclear factor/forkhead homologue 4 gene results in an absence of cilia and random left-right asymmetry. J. Clin. Invest. 102, 1077-1082.
Choksi, S.P., Lauter, G., Swoboda, P. and Roy, S. (2014). Switching on cilia: transcriptional networks regulating ciliogenesis. Development 141, 1427-1441.
Cruz, C., Ribes, V., Kutejova, E., Cayuso, J., Lawson, V., Norris, D., et al. (2010). Foxj1 regulates floor plate cilia architecture and modifies the response of cells to sonic hedgehog signalling. Development 137, 4271-4282.
Didon, L., Zwick, R.K., Chao, I.W., Walters, M.S., Wang, R., Hackett, N.R., et al. (2013). RFX3 Modulation of FOXJ1 regulation of cilia genes in the human airway epithelium. Respir. Res. 14, 70-70.
Gerovac, B.J. and Fregien, N.L. (2016). IL-13 inhibits multicilin expression and ciliogenesis via janus kinase/signal transducer and activator of transcription independently of Notch cleavage. Am. J. Respir. Cell Mol. Biol. 54, 554-561.
Gerovac, B.J., Valencia, M., Baumlin, N., Salathe, M., Conner, G.E. and Fregien, N.L. (2014). Submersion and hypoxia inhibit ciliated cell differentiation in a notch-dependent manner. Am. J. Respir. Cell Mol. Biol. 51(4), 516-525.
Gomperts, B.N., Gong-Cooper, X. and Hackett, B.P. (2004). Foxj1 regulates basal body anchoring to the cytoskeleton of ciliated pulmonary epithelial cells. J. Cell Sci. 117, 1329-1337.
Gomperts, B.N., Kim, L.J., Flaherty, S.A. and Hackett, B.P. (2007). IL-13 Regulates Cilia Loss and foxj1 Expression in Human Airway Epithelium. Am. J. Respir. Cell Mol. Biol. 37, 339-346.
Hackett, B.P., Brody, S.L., Liang, M., Zeitz, I.D., Bruns, L.A. and Gitlin, J.D. (1995). Primary structure of hepatocyte nuclear factor/forkhead homologue 4 and characterization of gene expression in the developing respiratory and reproductive epithelium. Proc. Natl. Acad. Sci. U. S. A. 92, 4249-4253.
Ishikawa, S. and Ito, S. (2017). Repeated whole cigarette smoke exposure alters cell differentiation and augments secretion of inflammatory mediators in air-liquid interface three-dimensional co-culture model of human bronchial tissue. Toxicol. in Vitro 38, 170-178.
Koc, M., Taysi, S., Buyukokuroglu, M.E. and Bakan, N. (2003). Melatonin protects rat liver against irradiation-induced oxidative injury. J. Radiat. Res. 44, 211-215.
Lim, L., Zhou, H. and Costa, R.H. (1997). The winged helix transcription factor HFH-4 is expressed during choroid plexus epithelial development in the mouse embryo. Proc. Natl. Acad. Sci. U. S. A. 94, 3094-3099.
Liu, M., Guan, Z., Shen, Q., Lalor, P., Fitzgerald, U., O'brien, T., et al., 2016. Ulk4 Is essential for ciliogenesis and CSF flow. J. Neurosci. 36, 7589-7600.
Milara, J., Armengot, M., Bañuls, P., Tenor, H., Beume, R., Artigues, E., et al. (2012). Roflumilast N-oxide, a PDE4 inhibitor, improves cilia motility and ciliated human bronchial epithelial cells compromised by cigarette smoke in vitro. Brit. J. Pharmacol. 166, 2243-2262.
Polosa, R., Emma, R., Cibella, F., Caruso, M., Conte, G., Benfatto, F., et al. (2021). Impact of exclusive e-cigarettes and heated tobacco products use on muco-ciliary clearance. Ther. Adv. Chronic Dis. 12, 20406223211035267-20406223211035267.
Rodrigues-Moreira, S., Moreno, S.G., Ghinatti, G., Lewandowski, D., Hoffschir, F., Ferri, F., et al. (2017). Low-Dose Irradiation Promotes Persistent Oxidative Stress and Decreases Self-Renewal in Hematopoietic Stem Cells. Cell Rep. 20, 3199-3211.
Schmid, A., Sailland, J., Novak, L., Baumlin, N., Fregien, N. and Salathe, M. (2017). Modulation of Wnt signaling is essential for the differentiation of ciliated epithelial cells in human airways. FEBS Lett. 591, 3493-3506.
Shaykhiev, R., Zuo, W.L., Chao, I., Fukui, T., Witover, B., Brekman, A., et al. (2013). EGF shifts human airway basal cell fate toward a smoking-associated airway epithelial phenotype. Proc. Natl. Acad. Sci. U. S. A. 110, 12102-12107.
Shirazi, A., Mihandoost, E., Ghobadi, G., Mohseni, M. and Ghazi-Khansari, M. (2013). Evaluation of radio-protective effect of melatonin on whole body irradiation induced liver tissue damage. Cell J. 14, 292-297.
Stauber, M., Weidemann, M., Dittrich-Breiholz, O., Lobschat, K., Alten, L., Mai, M., et al. (2017). Identification of FOXJ1 effectors during ciliogenesis in the foetal respiratory epithelium and embryonic left-right organiser of the mouse. Dev. Biol. 423, 170-188.
Stubbs, J.L., Vladar, E.K., Axelrod, J.D. and Kintner, C. (2012). Multicilin promotes centriole assembly and ciliogenesis during multiciliate cell differentiation. Nat. Cell Biol. 14, 140-147.
Tan, F.E., Vladar, E.K., Ma, L., Fuentealba, L.C., Hoh, R., Espinoza, F.H., et al. (2013). Myb promotes centriole amplification and later steps of the multiciliogenesis program. Development 140, 4277-4286.
Valencia-Gattas, M., Conner, G.E. and Fregien, N.L. (2016). Gefitinib, an EGFR Tyrosine Kinase inhibitor, Prevents Smoke-Mediated Ciliated Airway Epithelial Cell Loss and Promotes Their Recovery. PloS ONE 11, e0160216.
Vij, S., Rink, J.C., Ho, H.K., Babu, D., Eitel, M., Narasimhan, V., et al. (2012). Evolutionarily ancient association of the FoxJ1 transcription factor with the motile ciliogenic program. PLoS Genet. 8, e1003019.
Yu, X., Ng, C.P., Habacher, H. and Roy, S. (2008). Foxj1 transcription factors are master regulators of the motile ciliogenic program. Nat. Genet. 40, 1445-1453.
Zhou, F. and Roy, S. (2015). SnapShot: Motile Cilia. Cell 162, 224-224 e221.