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Smooth muscle CaMKIIδ promotes allergen-induced airway hyperresponsiveness and inflammation

  • Molecular and cellular mechanisms of disease
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Abstract

Airway smooth muscle (ASM) is a key target cell in allergen-induced asthma known to contribute to airway hyperresponsiveness (AHR) and chronic airway remodeling. Changes in ASM calcium homeostasis have been shown to contribute to AHR although the mechanisms and Ca2+ signal effectors are incompletely understood. In the present study, we tested the function of ASM multifunctional protein kinase Ca2+/calmodulin-dependent kinase II (CaMKII) isoforms CaMKIIδ and CaMKIIγ in allergen-induced AHR and airway remodeling in vivo. Using a murine model of atopic asthma, we demonstrate that CaMKIIδ protein is upregulated in ASM derived from ovalbumin (OVA)-treated animals compared to controls. A genetic approach to conditionally knock out smooth muscle CaMKIIδ and CaMKIIγ in separate Cre-loxp systems was validated, and using this loss-of-function approach, the function of these CaMKII isoforms was tested in ovalbumin (OVA)-induced airway remodeling and AHR. OVA treatment in control mice had no effect on ASM remodeling in this model of AHR, and CaMKIIδ knockouts had no independent effects on ASM content. However, at 1 day post-final OVA challenge, OVA-induced AHR was eliminated in the CaMKIIδ knockouts. OVA-induced peribronchial inflammation and bronchoalveolar lavage fluid (BALF) levels of the Th2 cytokine IL-13 were significantly decreased in the CaMKIIδ knockouts. Unexpectedly, we found increased peribronchial eosinophils in the smooth muscle CaMKIIδ knockouts compared to control animals at 1 day post-final challenge, suggesting that lack of ASM CaMKIIδ delays the progression of AHR rather than inhibiting it. Indeed, when AHR was determined at 7 days post-final OVA challenge, CaMKIIδ knockouts showed robust AHR while AHR was fully resolved in OVA-challenged control mice. These in vivo studies demonstrate a role for smooth muscle CaMKIIδ in promoting airway inflammation and AHR and suggest a complex signaling role for CaMKIIδ in regulating ASM function. These studies confirm the diverse roles of ASM cells as immune effectors that control AHR and call for further studies into CaMKIIδ-mediated signaling in ASM cells during disease.

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References

  1. Ammit AJ, Moir LM, Oliver BG, Hughes JM, Alkhouri H, Ge Q, Burgess JK, Black JL, Roth M (2007) Effect of IL-6 trans-signaling on the pro-remodeling phenotype of airway smooth muscle. Am J Physiol Lung Cell Mol Physiol 292:L199–L206

    Article  CAS  PubMed  Google Scholar 

  2. Amrani Y, Lazaar AL, Hoffman R, Amin K, Ousmer S, Panettieri RA Jr (2000) Activation of p55 tumor necrosis factor-alpha receptor-1 coupled to tumor necrosis factor receptor-associated factor 2 stimulates intercellular adhesion molecule-1 expression by modulating a thapsigargin-sensitive pathway in human tracheal smooth muscle cells. Mol Pharmacol 58:237–245

    CAS  PubMed  Google Scholar 

  3. Amrani Y, Panettieri RA (2003) Airway smooth muscle: contraction and beyond. Int J Biochem Cell Biol 35:272–276

    Article  CAS  PubMed  Google Scholar 

  4. An SS, Bai TR, Bates JH, Black JL, Brown RH, Brusasco V, Chitano P, Deng L, Dowell M, Eidelman DH, Fabry B et al (2007) Airway smooth muscle dynamics: a common pathway of airway obstruction in asthma. Eur Respir J 29:834–860

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  5. Backs J, Backs T, Neef S, Kreusser MM, Lehmann LH, Patrick DM, Grueter CE, Qi X, Richardson JA, Hill JA, Katus HA et al (2009) The delta isoform of CaM kinase II is required for pathological cardiac hypertrophy and remodeling after pressure overload. Proc Natl Acad Sci U S A 106:2342–2347

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  6. Backs J, Stein P, Backs T, Duncan FE, Grueter CE, McAnally J, Qi X, Schultz RM, Olson EN (2010) The gamma isoform of CaM kinase II controls mouse egg activation by regulating cell cycle resumption. Proc Natl Acad Sci U S A 107:81–86

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  7. Berkman N, Krishnan VL, Gilbey T, Newton R, O’Connor B, Barnes PJ, Chung KF (1996) Expression of RANTES mRNA and protein in airways of patients with mild asthma. Am J Respir Crit Care Med 154:1804–1811

    Article  CAS  PubMed  Google Scholar 

  8. Camoretti-Mercado B, Forsythe SM, LeBeau MM, Espinosa R III, Vieira JE, Halayko AJ, Willadsen S, Kurtz B, Ober C, Evans GA, Thweatt R et al (1998) Expression and cytogenetic localization of the human SM22 gene (TAGLN). Genomics 49:452–457

    Article  CAS  PubMed  Google Scholar 

  9. Catley MC, Sukkar MB, Chung KF, Jaffee B, Liao SM, Coyle AJ, Haddad E, Barnes PJ, Newton R (2006) Validation of the anti-inflammatory properties of small-molecule IkappaB Kinase (IKK)-2 inhibitors by comparison with adenoviral-mediated delivery of dominant-negative IKK1 and IKK2 in human airways smooth muscle. Mol Pharmacol 70:697–705

    Article  CAS  PubMed  Google Scholar 

  10. Cazzola M, Page CP, Rogliani P, Matera MG (2013) beta2-agonist therapy in lung disease. Am J Respir Crit Care Med 187:690–696

    Article  CAS  PubMed  Google Scholar 

  11. Cox G, Thomson NC, Rubin AS, Niven RM, Corris PA, Siersted HC, Olivenstein R, Pavord ID, McCormack D, Chaudhuri R, Miller JD et al (2007) Asthma control during the year after bronchial thermoplasty. N Engl J Med 356:1327–1337

    Article  CAS  PubMed  Google Scholar 

  12. Del Prete GF, De CM, D’Elios MM, Maestrelli P, Ricci M, Fabbri L, Romagnani S (1993) Allergen exposure induces the activation of allergen-specific Th2 cells in the airway mucosa of patients with allergic respiratory disorders. Eur J Immunol 23:1445–1449

    Article  PubMed  Google Scholar 

  13. Doherty TA, Soroosh P, Broide DH, Croft M (2009) CD4+ cells are required for chronic eosinophilic lung inflammation but not airway remodeling. Am J Physiol Lung Cell Mol Physiol 296:L229–L235

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  14. Elias JA, Wu Y, Zheng T, Panettieri R (1997) Cytokine- and virus-stimulated airway smooth muscle cells produce IL-11 and other IL-6-type cytokines. Am J Physiol 273:L648–L655

    CAS  PubMed  Google Scholar 

  15. Erjefalt JS, Persson CG (2000) New aspects of degranulation and fates of airway mucosal eosinophils. Am J Respir Crit Care Med 161:2074–2085

    Article  CAS  PubMed  Google Scholar 

  16. Frischauf I, Schindl R, Derler I, Bergsmann J, Fahrner M, Romanin C (2008) The STIM/Orai coupling machinery. Channels (Austin) 2:261–268

    Article  Google Scholar 

  17. Gunst SJ, Panettieri RA Jr (2012) Last Word on Point: alterations in airway smooth muscle phenotype do cause airway hyperresponsiveness in asthma. J Appl Physiol 113:847

    Article  PubMed  Google Scholar 

  18. Hakonarson H, Maskeri N, Carter C, Grunstein MM (1999) Regulation of TH1- and TH2-type cytokine expression and action in atopic asthmatic sensitized airway smooth muscle. J Clin Invest 103:1077–1087

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  19. Halayko AJ, Solway J (2001) Molecular mechanisms of phenotypic plasticity in smooth muscle cells. J Appl Physiol 90:358–368

    CAS  PubMed  Google Scholar 

  20. House SJ, Ginnan RG, Armstrong SE, Singer HA (2007) Calcium/calmodulin-dependent protein kinase II-delta isoform regulation of vascular smooth muscle cell proliferation. Am J Physiol Cell Physiol 292:C2276–C2287

    Article  CAS  PubMed  Google Scholar 

  21. House SJ, Singer HA (2008) CaMKII-delta isoform regulation of neointima formation after vascular injury. Arterioscler Thromb Vasc Biol 28:441–447

    Article  CAS  PubMed  Google Scholar 

  22. Hudmon A, Schulman H (2002) Structure-function of the multifunctional Ca2+/calmodulin-dependent protein kinase II. Biochem J 364:593–611

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  23. Jarai G, Sukkar M, Garrett S, Duroudier N, Westwick J, Adcock I, Chung KF (2004) Effects of interleukin-1beta, interleukin-13 and transforming growth factor-beta on gene expression in human airway smooth muscle using gene microarrays. Eur J Pharmacol 497:255–265

    Article  CAS  PubMed  Google Scholar 

  24. Kips JC (2003) The relation between morphologic and functional airway changes in bronchial asthma. Verh K Acad Geneeskd Belg 65:247–265

    CAS  PubMed  Google Scholar 

  25. Lamkhioued B, Renzi PM, Bi-Younes S, Garcia-Zepada EA, Allakhverdi Z, Ghaffar O, Rothenberg MD, Luster AD, Hamid Q (1997) Increased expression of eotaxin in bronchoalveolar lavage and airways of asthmatics contributes to the chemotaxis of eosinophils to the site of inflammation. J Immunol 159:4593–4601

    CAS  PubMed  Google Scholar 

  26. Lepore JJ, Cheng L, Min LM, Mericko PA, Morrisey EE, Parmacek MS (2005) High-efficiency somatic mutagenesis in smooth muscle cells and cardiac myocytes in SM22alpha-Cre transgenic mice. Genesis 41:179–184

    Article  CAS  PubMed  Google Scholar 

  27. Li W, Li H, Sanders PN, Mohler PJ, Backs J, Olson EN, Anderson ME, Grumbach IM (2011) The multifunctional Ca2+/calmodulin-dependent kinase II delta (CaMKIIdelta) controls neointima formation after carotid ligation and vascular smooth muscle cell proliferation through cell cycle regulation by p21. J Biol Chem 286:7990–7999

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  28. Li L, Miano JM, Mercer B, Olson EN (1996) Expression of the SM22alpha promoter in transgenic mice provides evidence for distinct transcriptional regulatory programs in vascular and visceral smooth muscle cells. J Cell Biol 132:849–859

    Article  CAS  PubMed  Google Scholar 

  29. Ling H, Gray CB, Zambon AC, Grimm M, Gu Y, Dalton N, Purcell NH, Peterson K, Brown JH (2013) Ca2+/Calmodulin-dependent protein kinase II delta mediates myocardial ischemia/reperfusion injury through nuclear factor-kappaB. Circ Res 112:935–944

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  30. Luo SF, Chang CC, Lee IT, Lee CW, Lin WN, Lin CC, Yang CM (2009) Activation of ROS/NF-kappaB and Ca2+/CaM kinase II are necessary for VCAM-1 induction in IL-1beta-treated human tracheal smooth muscle cells. Toxicol Appl Pharmacol 237:8–21

    Article  CAS  PubMed  Google Scholar 

  31. Mahn K, Hirst SJ, Ying S, Holt MR, Lavender P, Ojo OO, Siew L, Simcock DE, McVicker CG, Kanabar V, Snetkov VA et al (2009) Diminished sarco/endoplasmic reticulum Ca2+ ATPase (SERCA) expression contributes to airway remodelling in bronchial asthma. Proc Natl Acad Sci U S A 106:10775–10780

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  32. Malm-Erjefalt M, Greiff L, Ankerst J, Andersson M, Wallengren J, Cardell LO, Rak S, Persson CG, Erjefalt JS (2005) Circulating eosinophils in asthma, allergic rhinitis, and atopic dermatitis lack morphological signs of degranulation. Clin Exp Allergy 35:1334–1340

    Article  CAS  PubMed  Google Scholar 

  33. Mercure MZ, Ginnan R, Singer HA (2008) CaM kinase II delta2-dependent regulation of vascular smooth muscle cell polarization and migration. Am J Physiol Cell Physiol 294:C1465–C1475

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  34. Moessler H, Mericskay M, Li Z, Nagl S, Paulin D, Small JV (1996) The SM 22 promoter directs tissue-specific expression in arterial but not in venous or visceral smooth muscle cells in transgenic mice. Development 122:2415–2425

    CAS  PubMed  Google Scholar 

  35. Nials AT, Uddin S (2008) Mouse models of allergic asthma: acute and chronic allergen challenge. Dis Model Mech 1:213–220

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  36. Odaka M, Matsukura S, Kuga H, Kokubu F, Kasama T, Kurokawa M, Kawaguchi M, Ieki K, Suzuki S, Watanabe S, Homma T et al (2007) Differential regulation of chemokine expression by Th1 and Th2 cytokines and mechanisms of eotaxin/CCL-11 expression in human airway smooth muscle cells. Int Arch Allergy Immunol 143(Suppl 1):84–88

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  37. Owens GK (1998) Molecular control of vascular smooth muscle cell differentiation. Acta Physiol Scand 164:623–635

    Article  CAS  PubMed  Google Scholar 

  38. Owens GK, Kumar MS, Wamhoff BR (2004) Molecular regulation of vascular smooth muscle cell differentiation in development and disease. Physiol Rev 84:767–801

    Article  CAS  PubMed  Google Scholar 

  39. Pare PD, Mitzner W (2012) Last Word on Counterpoint: alterations in airway smooth muscle phenotype do not cause airway hyperresponsiveness in asthma. J Appl Physiol 113:848

    Article  PubMed Central  PubMed  Google Scholar 

  40. Pepe C, Foley S, Shannon J, Lemiere C, Olivenstein R, Ernst P, Ludwig MS, Martin JG, Hamid Q (2005) Differences in airway remodeling between subjects with severe and moderate asthma. J Allergy Clin Immunol 116:544–549

    Article  PubMed  Google Scholar 

  41. Prefontaine D, Lajoie-Kadoch S, Foley S, Audusseau S, Olivenstein R, Halayko AJ, Lemiere C, Martin JG, Hamid Q (2009) Increased expression of IL-33 in severe asthma: evidence of expression by airway smooth muscle cells. J Immunol 183:5094–5103

    Article  CAS  PubMed  Google Scholar 

  42. Ramos-Barbon D, Fraga-Iriso R, Brienza NS, Montero-Martinez C, Verea-Hernando H, Olivenstein R, Lemiere C, Ernst P, Hamid QA, Martin JG (2010) T Cells localize with proliferating smooth muscle alpha-actin + cell compartments in asthma. Am J Respir Crit Care Med 182:317–324

    Article  PubMed  Google Scholar 

  43. Robins S, Roussel L, Schachter A, Risse PA, Mogas AK, Olivenstein R, Martin JG, Hamid Q, Rousseau S (2011) Steroid-insensitive ERK1/2 activity drives CXCL8 synthesis and neutrophilia by airway smooth muscle. Am J Respir Cell Mol Biol 45:984–990

    Article  CAS  PubMed  Google Scholar 

  44. Sanders PN, Koval OM, Jaffer OA, Prasad AM, Businga TR, Scott JA, Hayden PJ, Luczak ED, Dickey DD, Allamargot C, Olivier AK et al (2013) CaMKII is essential for the proasthmatic effects of oxidation. Sci Transl Med 5:195ra97

    Article  PubMed Central  PubMed  Google Scholar 

  45. Saunders MA, Mitchell JA, Seldon PM, Yacoub MH, Barnes PJ, Giembycz MA, Belvisi MG (1997) Release of granulocyte-macrophage colony stimulating factor by human cultured airway smooth muscle cells: suppression by dexamethasone. Br J Pharmacol 120:545–546

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  46. Schmitz J, Owyang A, Oldham E, Song Y, Murphy E, McClanahan TK, Zurawski G, Moshrefi M, Qin J, Li X, Gorman DM et al (2005) IL-33, an interleukin-1-like cytokine that signals via the IL-1 receptor-related protein ST2 and induces T helper type 2-associated cytokines. Immunity 23:479–490

    Article  CAS  PubMed  Google Scholar 

  47. Schramm CM, Puddington L, Wu C, Guernsey L, Gharaee-Kermani M, Phan SH, Thrall RS (2004) Chronic inhaled ovalbumin exposure induces antigen-dependent but not antigen-specific inhalational tolerance in a murine model of allergic airway disease. Am J Pathol 164:295–304

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  48. Schworer CM, Rothblum LI, Thekkumkara TJ, Singer HA (1993) Identification of novel isoforms of the delta subunit of Ca2+/calmodulin-dependent protein kinase II. Differential expression in rat brain and aorta. J Biol Chem 268:14443–14449

    CAS  PubMed  Google Scholar 

  49. Shan L, Redhu NS, Saleh A, Halayko AJ, Chakir J, Gounni AS (2010) Thymic stromal lymphopoietin receptor-mediated IL-6 and CC/CXC chemokines expression in human airway smooth muscle cells: role of MAPKs (ERK1/2, p38, and JNK) and STAT3 pathways. J Immunol 184:7134–7143

    Article  CAS  PubMed  Google Scholar 

  50. Shi HZ, Humbles A, Gerard C, Jin Z, Weller PF (2000) Lymph node trafficking and antigen presentation by endobronchial eosinophils. J Clin Invest 105:945–953

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  51. Spinelli AM, Gonzalez-Cobos JC, Zhang X, Motiani RK, Rowan S, Zhang W, Garrett J, Vincent PA, Matrougui K, Singer HA, Trebak M (2012) Airway smooth muscle STIM1 and Orai1 are upregulated in asthmatic mice and mediate PDGF-activated SOCE, CRAC currents, proliferation, and migration. Pflugers Arch 464:481–492

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  52. Sumi M, Kiuchi K, Ishikawa T, Ishii A, Hagiwara M, Nagatsu T, Hidaka H (1991) The newly synthesized selective Ca2+/calmodulin dependent protein kinase II inhibitor KN-93 reduces dopamine contents in PC12h cells. Biochem Biophys Res Commun 181:968–975

    Article  CAS  PubMed  Google Scholar 

  53. Tan X, Alrashdan YA, Alkhouri H, Oliver BG, Armour CL, Hughes JM (2013) Airway smooth muscle CXCR3 ligand production: regulation by JAK-STAT1 and intracellular Ca(2)(+). Am J Physiol Lung Cell Mol Physiol 304:L790–L802

    Article  CAS  PubMed  Google Scholar 

  54. Tombes RM, Faison MO, Turbeville JM (2003) Organization and evolution of multifunctional Ca(2+)/CaM-dependent protein kinase genes. Gene 322:17–31

    Article  CAS  PubMed  Google Scholar 

  55. Trian T, Benard G, Begueret H, Rossignol R, Girodet PO, Ghosh D, Ousova O, Vernejoux JM, Marthan R, Tunon-de-Lara JM, Berger P (2007) Bronchial smooth muscle remodeling involves calcium-dependent enhanced mitochondrial biogenesis in asthma. J Exp Med 204:3173–3181

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  56. Venge P (2010) The eosinophil and airway remodelling in asthma. Clin Respir J 4(Suppl 1):15–19

    Article  CAS  PubMed  Google Scholar 

  57. Walker C, Bode E, Boer L, Hansel TT, Blaser K, Virchow JC Jr (1992) Allergic and nonallergic asthmatics have distinct patterns of T-cell activation and cytokine production in peripheral blood and bronchoalveolar lavage. Am Rev Respir Dis 146:109–115

    Article  CAS  PubMed  Google Scholar 

  58. Wright DB, Trian T, Siddiqui S, Pascoe CD, Johnson JR, Dekkers BG, Dakshinamurti S, Bagchi R, Burgess JK, Kanabar V, Ojo OO (2013) Phenotype modulation of airway smooth muscle in asthma. Pulm Pharmacol Ther 26:42–49

    Article  CAS  PubMed  Google Scholar 

  59. Zuyderduyn S, Sukkar MB, Fust A, Dhaliwal S, Burgess JK (2008) Treating asthma means treating airway smooth muscle cells. Eur Respir J 32:265–274

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

The authors greatly acknowledge the following research support: AMC’s Candice Weir Fund, NIH grant HL097111 (to MT), NIH grant HL049426 (to HAS), and Albany Medical Center fellowship and stipend support (AMS). We would also like to acknowledge receipt of generous gifts including founder mice carry floxed CaMKIIδ and CaMKIIγ alleles which were kindly provided by Dr. Johannes Backs (Heidelberg) and Dr. Eric Olson (UTSW) and the eosinophil-specific MPB antibody, provided by Dr. Jamie Lee (Mayo Clinic Scottsdale, Arizona). We also thank Xiaolan Ding, Miao Jiang, and Diane Singer for their general animal and cell culture support, Dr. Margarida Barroso for her assistance and insight into immunohistochemical and microscopy techniques, and the Singer, Trebak, and Jourd’heuil lab for the exchange of ideas and experiences on the first floor of the ME building at AMC.

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  1. Amy M. Spinelli

    Present address: Department of Cellular and Molecular Physiology, The Pennsylvania State University College of Medicine, Hershey, PA, USA

  2. José C. González-Cobos

    Present address: Barlovento Brewing Company, Manati, Puerto Rico

Authors and Affiliations

  1. Center for Cardiovascular Sciences, Albany Medical College, Albany, NY, USA

    Amy M. Spinelli, Yongfeng Liu, Li-Yan Sun, José C. González-Cobos & Harold A. Singer

  2. Department of Cellular and Molecular Physiology, The Pennsylvania State University College of Medicine, Hershey, PA, USA

    Mohamed Trebak

  3. Laboratory for Cardiac Epigenetics, Department of Cardiology, University of Heidelberg, and German Center for Cardiovascular Research (DZHK), Heidelberg, Germany

    Johannes Backs

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  1. Amy M. Spinelli
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Spinelli, A.M., Liu, Y., Sun, LY. et al. Smooth muscle CaMKIIδ promotes allergen-induced airway hyperresponsiveness and inflammation. Pflugers Arch - Eur J Physiol 467, 2541–2554 (2015). https://doi.org/10.1007/s00424-015-1713-5

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