Allergy Asthma Immunol Res.  2012 Jul;4(4):222-230. 10.4168/aair.2012.4.4.222.

Roles of Periostin in Symptom Manifestation and Airway Remodeling in a Murine Model of Allergic Rhinitis

• Hur DG
• Khalmuratova R
• Ahn SK
• Ha YS
• Min YG

• KMID: 1970711
• DOI: http://doi.org/10.4168/aair.2012.4.4.222

Abstract

PURPOSE
Periostin was originally identified as a secreted factor during screening of a mouse osteoblastic library. In a recent study, periostin was found to directly regulate eosinophil accumulation in allergic mucosal inflammation. Chronic eosinophilic inflammation is related to the development of remodeling. The present study examined the expression of periostin and evaluated its role in the inflammatory process and remodeling associated with allergic rhinitis.
METHODS
A murine model of allergic rhinitis was established in periostin knockout mice. We analyzed the expression of periostin, manifestation of nasal symptoms, eosinophilic inflammation, and subepithelial fibrosis as well as the expression of MMP-2, TIMP-1, and type 1 collagen in nasal tissue.
RESULTS
Periostin was mainly distributed in the subepithelial tissue of the nasal mucosa. The subepithelial tissue was thinner in the knockout group than in the control group. No differences in the expression of MMP-2 or TIMP-1 were found in the knockout group. However, after a month of allergen challenge, type I collagen in the nasal tissue was lower in the knockout group than in the control group. The number of eosinophils and the symptom score were also lower in the knockout group.
CONCLUSIONS
Periostin is expressed in nasal tissues of murine models of allergic rhinitis. Periostin deficiency may affect the remodeling of nasal tissue with reduced subepithelial fibrosis, and lead to less eosinophilic inflammation.

Keyword

Airway remodeling; allergy; eosinophils; mice; periostin protein; rhinitis

MeSH Terms

Airway Remodeling
Animals
Collagen Type I
Eosinophils
Fibrosis
Hypersensitivity
Inflammation
Mass Screening
Mice
Mice, Knockout
Nasal Mucosa
Osteoblasts
Rhinitis
Rhinitis, Allergic, Perennial
Tissue Inhibitor of Metalloproteinase-1
Collagen Type I
Tissue Inhibitor of Metalloproteinase-1

Figure

• 

  Fig. 1 Protocol for induction of the murine AR model. Three groups of mice were used in these experiments, and each group contained three subgroups, according to the day of sacrifice.PBS, phosphate buffered saline; OVA, ovalbumin; KO, knockout; W, week; M, month.

• 

  Fig. 2 Periostin immunohistochemistry. Images show the nasal respiratory mucosa after 3 months of allergen challenge. (A) OVA group, original magnification, ×20. (B) OVA group, original magnification, ×400. (C) PBS group, original magnification, ×400. (D) KO group, original magnification, ×400. Periostin was highly expressed in the subepithelial area in the OVA group (asterisk in B). However, it was less expressed in the same area of the PBS group, and no periostin expression was observed in the KO group.PBS, phosphate buffered saline; OVA, ovalbumin; KO, knockout.

• 

  Fig. 3 Periostin levels in serum. Serum periostin levels were not significantly different in the PBS or OVA groups over time. After 19 days, 1 month, and 3 months of allergen challenge, the PBS and OVA groups showed similar serum periostin levels. Serum periostin was not found in the KO group. Data are expressed as mean±SE.PBS, phosphate buffered saline; OVA, ovalbumin; KO, knockout.

• 

  Fig. 4 Subepithelial thickness of the nasal mucosa. The subepithelial thickness of the nasal mucosa was measured (yellow arrow). (A) The nasal tissue around the septal cartilage in the PBS group after 3 months of allergen challenge (Masson's trichrome staining, original magnification, ×400). (B) The same area in the OVA group after 3 months of allergen challenge. (C) The same area in the KO group after 3 months of allergen challenge. (D) The subepithelial tissue was thinner in the KO group than in the OVA groups. *P<0.05. The subepithelial thickness increased over time in the OVA group. **P<0.05. Data are expressed as mean±SE.PBS, phosphate buffered saline; OVA, ovalbumin; KO, knockout.

• 

  Fig. 5 Expression of type I collagen in the KO group visualized by immunohistochemistry. (A) Original magnification, ×20. (B) Original magnification, ×400. Type I collagen was identified in the subepithelial tissue (asterisk in B) of the nasal mucosa, regardless of periostin deficiency. The graph depicts the ratio of collagen type I to β-actin from nasal tissue lysates. The KO group showed a trend toward decreased type I collagen expression after 1 and 3 months of allergen challenge compared to the OVA group by immunoblotting. Two representative results of immunoblotting (n=5) over time are shown.OVA, ovalbumin; KO, knockout.

• 

  Fig. 6 MMP-2 and TIMP-1 expression in the nasal mucosa. The KO and OVA groups showed no significant differences in MMP-2 or TIMP-1 expression in the nasal mucosa by immunoblotting over time. MMP-2, matrix metalloproteinase 2; TIMP-1, tissue inhibitor of metalloproteinase. Two representative results of immunoblotting (n=5) over time are shown.OVA, ovalbumin; KO, knockout.

• 

  Fig. 7 Eosinophil counts in the nasal mucosa. The nasal tissue around conchal cartilage in the OVA group after (A) 19 days, (B) 1 month, and (C) 3 months are shown. Eosinophils are reddish (arrow). The nasal tissue of the KO group after (D) 19 days, (E) 1 month, and (F) 3 months are also shown (Luna staining, original magnification, ×400). (G) The number of eosinophils in the nasal mucosa was significantly lower in the KO group than in the OVA groups. *P<0.05. In the OVA group, the number of eosinophils was higher at 3 months of allergen challenge than at the other time points. **P<0.05. Data are expressed as mean±SE.HPF, high power field.

• 

  Fig. 8 Allergen-induced nasal symptoms. The numbers of (A) sneezes and (B) rubbings for 10 min immediately after the allergen challenge were counted. The OVA group had a higher symptom score than the PBS group at all time points, *P<0.05; The KO group had a lower sneeze score than the OVA group at 1 and 3 months of allergen challenge. The KO group had a lower rubbing score than the OVA group at 3 months of allergen challenge, **P<0.05. Data are expressed as mean±SE.

Reference

1. Humbles AA, Lloyd CM, McMillan SJ, Friend DS, Xanthou G, McKenna EE, Ghiran S, Gerard NP, Yu C, Orkin SH, Gerard C. A critical role for eosinophils in allergic airways remodeling. Science. 2004; 305:1776–1779. PMID: 15375268.

2. Jeffery PK. Remodeling and inflammation of bronchi in asthma and chronic obstructive pulmonary disease. Proc Am Thorac Soc. 2004; 1:176–183. PMID: 16113432.

3. Bousquet J, Van Cauwenberge P, Khaltaev N. Aria Workshop Group. World Health Organization. Allergic rhinitis and its impact on asthma. J Allergy Clin Immunol. 2001; 108:S147–S334. PMID: 11707753.

4. Bousquet J, Jacot W, Vignola AM, Bachert C, Van Cauwenberge P. Allergic rhinitis: a disease remodeling the upper airways? J Allergy Clin Immunol. 2004; 113:43–49. PMID: 14713906.

5. Butcher JT, Norris RA, Hoffman S, Mjaatvedt CH, Markwald RR. Periostin promotes atrioventricular mesenchyme matrix invasion and remodeling mediated by integrin signaling through Rho/PI 3-kinase. Dev Biol. 2007; 302:256–266. PMID: 17070513.

6. Takayama G, Arima K, Kanaji T, Toda S, Tanaka H, Shoji S, McKenzie AN, Nagai H, Hotokebuchi T, Izuhara K. Periostin: a novel component of subepithelial fibrosis of bronchial asthma downstream of IL-4 and IL-13 signals. J Allergy Clin Immunol. 2006; 118:98–104. PMID: 16815144.

7. Yuyama N, Davies DE, Akaiwa M, Matsui K, Hamasaki Y, Suminami Y, Yoshida NL, Maeda M, Pandit A, Lordan JL, Kamogawa Y, Arima K, Nagumo F, Sugimachi M, Berger A, Richards I, Roberds SL, Yamashita T, Kishi F, Kato H, Arai K, Ohshima K, Tadano J, Hamasaki N, Miyatake S, Sugita Y, Holgate ST, Izuhara K. Analysis of novel disease-related genes in bronchial asthma. Cytokine. 2002; 19:287–296. PMID: 12421571.

8. Horiuchi K, Amizuka N, Takeshita S, Takamatsu H, Katsuura M, Ozawa H, Toyama Y, Bonewald LF, Kudo A. Identification and characterization of a novel protein, periostin, with restricted expression to periosteum and periodontal ligament and increased expression by transforming growth factor beta. J Bone Miner Res. 1999; 14:1239–1249. PMID: 10404027.

9. Huber O, Sumper M. Algal-CAMs: isoforms of a cell adhesion molecule in embryos of the alga Volvox with homology to Drosophila fasciclin I. EMBO J. 1994; 13:4212–4222. PMID: 7925267.

10. Litvin J, Selim AH, Montgomery MO, Lehmann K, Rico MC, Devlin H, Bednarik DP, Safadi FF. Expression and function of periostin-isoforms in bone. J Cell Biochem. 2004; 92:1044–1061. PMID: 15258926.

11. Skonier J, Neubauer M, Madisen L, Bennett K, Plowman GD, Purchio AF. cDNA cloning and sequence analysis of beta ig-h3, a novel gene induced in a human adenocarcinoma cell line after treatment with transforming growth factor-beta. DNA Cell Biol. 1992; 11:511–522. PMID: 1388724.

12. Terasaka K, Yamaguchi R, Matsuo K, Yamazaki A, Nagai S, Yamada T. Complete nucleotide sequence of immunogenic protein MPB70 from Mycobacterium bovis BCG. FEMS Microbiol Lett. 1989; 49:273–276. PMID: 2663636.

13. Bao S, Ouyang G, Bai X, Huang Z, Ma C, Liu M, Shao R, Anderson RM, Rich JN, Wang XF. Periostin potently promotes metastatic growth of colon cancer by augmenting cell survival via the Akt/PKB pathway. Cancer Cell. 2004; 5:329–339. PMID: 15093540.

14. Gonzalez HE, Gujrati M, Frederick M, Henderson Y, Arumugam J, Spring PW, Mitsudo K, Kim HW, Clayman GL. Identification of 9 genes differentially expressed in head and neck squamous cell carcinoma. Arch Otolaryngol Head Neck Surg. 2003; 129:754–759. PMID: 12874078.

15. Kudo Y, Siriwardena BS, Hatano H, Ogawa I, Takata T. Periostin: novel diagnostic and therapeutic target for cancer. Histol Histopathol. 2007; 22:1167–1174. PMID: 17616943.

16. Norris RA, Moreno-Rodriguez RA, Sugi Y, Hoffman S, Amos J, Hart MM, Potts JD, Goodwin RL, Markwald RR. Periostin regulates atrioventricular valve maturation. Dev Biol. 2008; 316:200–213. PMID: 18313657.

17. Sasaki H, Yu CY, Dai M, Tam C, Loda M, Auclair D, Chen LB, Elias A. Elevated serum periostin levels in patients with bone metastases from breast but not lung cancer. Breast Cancer Res Treat. 2003; 77:245–252. PMID: 12602924.

18. Blanchard C, Mingler MK, McBride M, Putnam PE, Collins MH, Chang G, Stringer K, Abonia JP, Molkentin JD, Rothenberg ME. Periostin facilitates eosinophil tissue infiltration in allergic lung and esophageal responses. Mucosal Immunol. 2008; 1:289–296. PMID: 19079190.

19. Nakaya M, Dohi M, Okunishi K, Nakagome K, Tanaka R, Imamura M, Yamamoto K, Kaga K. Prolonged allergen challenge in murine nasal allergic rhinitis: nasal airway remodeling and adaptation of nasal airway responsiveness. Laryngoscope. 2007; 117:881–885. PMID: 17473688.

20. Kitajima S, Kudo Y, Ogawa I, Bashir T, Kitagawa M, Miyauchi M, Pagano M, Takata T. Role of Cks1 overexpression in oral squamous cell carcinomas: cooperation with Skp2 in promoting p27 degradation. Am J Pathol. 2004; 165:2147–2155. PMID: 15579456.

21. Silberstein DS. Eosinophil function in health and disease. Crit Rev Oncol Hematol. 1995; 19:47–77. PMID: 7741980.

22. Bavbek S, Sencer H, Misirligil Z, Beder S, Gurbuz L. Light and electron microscope study in allergic rhinitis patients (ARP) with or without bronchial hyperreactivity (BHR). J Investig Allergol Clin Immunol. 1996; 6:172–182.

23. Sanai A, Nagata H, Konno A. Extensive interstitial collagen deposition on the basement membrane zone in allergic nasal mucosa. Acta Otolaryngol. 1999; 119:473–478. PMID: 10445064.

24. Berry M, Morgan A, Shaw DE, Parker D, Green R, Brightling C, Bradding P, Wardlaw AJ, Pavord ID. Pathological features and inhaled corticosteroid response of eosinophilic and non-eosinophilic asthma. Thorax. 2007; 62:1043–1049. PMID: 17356056.

25. Brown NJ, Salome CM, Berend N, Thorpe CW, King GG. Airway distensibility in adults with asthma and healthy adults, measured by forced oscillation technique. Am J Respir Crit Care Med. 2007; 176:129–137. PMID: 17463413.

26. Ward C, Johns DP, Bish R, Pais M, Reid DW, Ingram C, Feltis B, Walters EH. Reduced airway distensibility, fixed airflow limitation, and airway wall remodeling in asthma. Am J Respir Crit Care Med. 2001; 164:1718–1721. PMID: 11719315.

27. Brewster CE, Howarth PH, Djukanovic R, Wilson J, Holgate ST, Roche WR. Myofibroblasts and subepithelial fibrosis in bronchial asthma. Am J Respir Cell Mol Biol. 1990; 3:507–511. PMID: 2223105.

28. Evans MJ, Fanucchi MV, Baker GL, Van Winkle LS, Pantle LM, Nishio SJ, Schelegle ES, Gershwin LJ, Miller LA, Hyde DM, Plopper CG. The remodelled tracheal basement membrane zone of infant rhesus monkeys after 6 months of recovery. Clin Exp Allergy. 2004; 34:1131–1136. PMID: 15248862.

29. Sidhu SS, Yuan S, Innes AL, Kerr S, Woodruff PG, Hou L, Muller SJ, Fahy JV. Roles of epithelial cell-derived periostin in TGF-beta activation, collagen production, and collagen gel elasticity in asthma. Proc Natl Acad Sci U S A. 2010; 107:14170–14175. PMID: 20660732.

30. Blyth DI, Wharton TF, Pedrick MS, Savage TJ, Sanjar S. Airway subepithelial fibrosis in a murine model of atopic asthma: suppression by dexamethasone or anti-interleukin-5 antibody. Am J Respir Cell Mol Biol. 2000; 23:241–246. PMID: 10919992.

31. Cho JY, Miller M, Baek KJ, Han JW, Nayar J, Lee SY, McElwain K, McElwain S, Friedman S, Broide DH. Inhibition of airway remodeling in IL-5-deficient mice. J Clin Invest. 2004; 113:551–560. PMID: 14966564.

32. Flood-Page PT, Menzies-Gow AN, Kay AB, Robinson DS. Eosinophil's role remains uncertain as anti-interleukin-5 only partially depletes numbers in asthmatic airway. Am J Respir Crit Care Med. 2003; 167:199–204. PMID: 12406833.

33. Wenzel SE, Schwartz LB, Langmack EL, Halliday JL, Trudeau JB, Gibbs RL, Chu HW. Evidence that severe asthma can be divided pathologically into two inflammatory subtypes with distinct physiologic and clinical characteristics. Am J Respir Crit Care Med. 1999; 160:1001–1008. PMID: 10471631.

34. Ohno I, Nitta Y, Yamauchi K, Hoshi H, Honma M, Woolley K, O'Byrne P, Tamura G, Jordana M, Shirato K. Transforming growth factor beta 1 (TGF beta 1) gene expression by eosinophils in asthmatic airway inflammation. Am J Respir Cell Mol Biol. 1996; 15:404–409. PMID: 8810646.

35. Rios H, Koushik SV, Wang H, Wang J, Zhou HM, Lindsley A, Rogers R, Chen Z, Maeda M, Kruzynska-Frejtag A, Feng JQ, Conway SJ. Periostin null mice exhibit dwarfism, incisor enamel defects, and an early-onset periodontal disease-like phenotype. Mol Cell Biol. 2005; 25:11131–11144. PMID: 16314533.

36. Takeshita S, Kikuno R, Tezuka K, Amann E. Osteoblast-specific factor 2: cloning of a putative bone adhesion protein with homology with the insect protein fasciclin I. Biochem J. 1993; 294:271–278. PMID: 8363580.

37. Kruzynska-Frejtag A, Machnicki M, Rogers R, Markwald RR, Conway SJ. Periostin (an osteoblast-specific factor) is expressed within the embryonic mouse heart during valve formation. Mech Dev. 2001; 103:183–188. PMID: 11335131.

38. Kudo H, Amizuka N, Araki K, Inohaya K, Kudo A. Zebrafish periostin is required for the adhesion of muscle fiber bundles to the myoseptum and for the differentiation of muscle fibers. Dev Biol. 2004; 267:473–487. PMID: 15013807.

39. Katsuragi N, Morishita R, Nakamura N, Ochiai T, Taniyama Y, Hasegawa Y, Kawashima K, Kaneda Y, Ogihara T, Sugimura K. Periostin as a novel factor responsible for ventricular dilation. Circulation. 2004; 110:1806–1813. PMID: 15381649.

40. Blanchard C, Wang N, Stringer KF, Mishra A, Fulkerson PC, Abonia JP, Jameson SC, Kirby C, Konikoff MR, Collins MH, Cohen MB, Akers R, Hogan SP, Assa'ad AH, Putnam PE, Aronow BJ, Rothenberg ME. Eotaxin-3 and a uniquely conserved gene-expression profile in eosinophilic esophagitis. J Clin Invest. 2006; 116:536–547. PMID: 16453027.

41. Sehra S, Yao W, Nguyen ET, Ahyi AN, Barbe Tuana FM, Ahlfeld SK, Snider P, Tepper RS, Petrache I, Conway SJ, Kaplan MH. Periostin regulates goblet cell metaplasia in a model of allergic airway inflammation. J Immunol. 2011; 186:4959–4966. PMID: 21402898.