Molecular Biology Reports

, Volume 40, Issue 11, pp 6091–6096

Adenovirus conducted connective tissue growth factor on extracellular matrix in trabecular meshwork and its role on aqueous humor outflow facility

  • Ying Su
  • Jingli Cheng
  • Hongtao Liu
  • Feng Wang
  • Shiguang Zhao
Article

Abstract

Deposition of extracellular matrix (ECM) in trabecular meshwork, such as fibronectin, collagen IV, elastin. leads to increased resistance of trabecular meshwork in primary open angle glaucoma (POAG). Connective tissue growth factor (CTGF) is known to regulate the ECM deposits. In this study, we detect the effect of adenovirus conducted CTGF (Adv-CTGF) transfection on either the expression of ECM components or aqueous humor outflow facility. Adv-CTGF was used to transfect rat trabecular meshwork cells in vivo and in vitro. Aqueous humor outflow facility was test by microbeads perfusion. Protein expression of CTGF, fibronectin, and collagen IV was determined using Western blot. In the Adv-CTGF group, the outflow facility displayed a significant decrease from baseline. It appears as though the transfection with Adv-CTGF significantly affects the aqueous humor outflow pattern. A negative correlation between IOP and PEFL indicated that a decrease in the area of bead deposition corresponded to an overall decrease of outflow, leading to an elevated IOP. Adv-CTGF can enhance the expression of CTGF, fibronectin and collagen IV. CTGF is the novel target for treatment of POAG. It is necessary to further study to test inhibition of CTGF expression for treatment of POAG.

Keywords

Glaucoma Trabecular meshwork Connective tissue growth factor Microbead Facility 

References

  1. 1.
    Quigley HA, Broman AT (2006) The number of people with glaucoma worldwide in 2010 and 2020. Br J Ophthalmol 90:262–267PubMedCrossRefGoogle Scholar
  2. 2.
    Lutjen-Drecoll E (2005) Morphological changes in glaucomatous eyes and the role of TGFβ2 for the pathogenesis of the disease. Exp Eye Res 81:1–4PubMedCrossRefGoogle Scholar
  3. 3.
    Hynes RO (2009) The extracellular matrix: not just pretty fibrils. Science 326:1216–1219PubMedCrossRefGoogle Scholar
  4. 4.
    Chung EJ, Lee HK, Jung SA, Lee SJ, Chee HY, Sohn YH, Lee JH (2012) Transduction of PTEN proteins using the tat domain modulates TGF-β1-mediated signaling pathways and transdifferentiation in subconjunctival fibroblasts. Invest Ophthalmol Vis Sci 53(1):379–386PubMedCrossRefGoogle Scholar
  5. 5.
    Takai Y, Tanito M, Ohira A (2012) Multiplex cytokine analysis of aqueous humor in eyes with primary open-angle glaucoma, exfoliation glaucoma, and cataract. Invest Ophthalmol Vis Sci 53(1):241–247PubMedCrossRefGoogle Scholar
  6. 6.
    Zenkel M, Krysta A, Pasutto F, Juenemann A, Kruse FE, Schlötzer-Schrehardt U (2011) Regulation of lysyl oxidase-like 1 (LOXL1) and elastin-related genes by pathogenic factors associated with pseudoexfoliation syndrome. Invest Ophthalmol Vis Sci 52(11):8488–8495PubMedCrossRefGoogle Scholar
  7. 7.
    Tektas OY, Lutjen-Drecoll E (2009) Structural changes of the trabecular meshwork in different kinds of glaucoma. Exp Eye Res 88:769–775PubMedCrossRefGoogle Scholar
  8. 8.
    Roberts AB, Heine UI, Flanders KC, Sporn MB (1990) Transforming growth factor-β. Major role in regulation of extracellular matrix. Ann N Y Acad Sci 580:225–232PubMedCrossRefGoogle Scholar
  9. 9.
    Bollinger KE, Crabb JS, Yuan X, Putliwala T, Clark AF, Crabb JW (2011) Quantitative proteomics: TGFβ2 signaling in trabecular meshwork cells. Invest Ophthalmol Vis Sci 52(11):8287–8294PubMedCrossRefGoogle Scholar
  10. 10.
    Read AT, Chan DW, Ethier CR (2007) Actin structure in the outflow tract of normal and glaucomatous eyes. Exp Eye Res 84:214–226PubMedCrossRefGoogle Scholar
  11. 11.
    Yam GH, Gaplovska-Kysela K, Zuber C, Roth J (2007) Sodium 4-phenylbutyrate acts as a chemical chaperone on misfolded myocilin to rescue cells from endoplasmic reticulum stress and apoptosis. Invest Ophthalmol Vis Sci 48:1683–1690PubMedCrossRefGoogle Scholar
  12. 12.
    Jacobson N, Andrews M, Shepard AR et al (2001) Non-secretion of mutant proteins of the glaucoma gene myocilin in cultured trabecular meshwork cells and in aqueous humor. Hum Mol Genet 10:117–125PubMedCrossRefGoogle Scholar
  13. 13.
    Ihn H (2002) Pathogenesis of fibrosis: role of TGF-β and CTGF. Curr Opin Rheumatol 14:681–685PubMedCrossRefGoogle Scholar
  14. 14.
    Leask A, Abraham DJ (2004) TGF-β signaling and the fibrotic response. FASEB J 18:816–827PubMedCrossRefGoogle Scholar
  15. 15.
    Ying Su, Yang Chen-Yuan C, Li Zhongrui, Feng Xu, Zhang Lei, Wang Feng, Zhao Shiguang (2012) Smad7 siRNA inhibit expression of extracellular matrix in trabecular meshwork cells treated with TGF-β2. Mol Vis 18:1881–1884Google Scholar
  16. 16.
    Buckingham BP, Inman DM, Lambert W, Oglesby E, Calkins DJ, Steele MR, Vetter ML, Marsh-Armstrong N, Horner PJ (2008) Progressive ganglion cell degeneration precedes neuronal loss in a mouse model of glaucoma. J Neurosci 28:2735–2744PubMedCrossRefGoogle Scholar
  17. 17.
    Howell GR, Libby RT, Jakobs TC, Smith RS, Phalan FC, Barter JW, Barbay JM, Marchant JK, Mahesh N, Porciatti V, Whitmore AV, Masland RH, John SW (2007) Axons of retinal ganglion cells are insulted in the optic nerve early in DBA/2J glaucoma. J Cell Biol 179:1523–1537PubMedCrossRefGoogle Scholar
  18. 18.
    Chudgar SM, Deng P, Maddala R, Epstein DL, Rao PV (2006) Regulation of connective tissue growth factor expression in the aqueous humor outflow pathway. Mol Vis 12:1117–1126PubMedGoogle Scholar
  19. 19.
    Junglas B, Yu AH, Welge-Lussen U, Tamm ER, Fuchshofer R (2009) Connective tissue growth factor induces extracellular matrix deposition in human trabecular meshwork cells. Exp Eye Res 88:1065–1075PubMedCrossRefGoogle Scholar
  20. 20.
    Ihn H (2002) Pathogenesis of fibrosis: role of TGF-βand CTGF. Curr Opin Rheumatol 14:681–685PubMedCrossRefGoogle Scholar
  21. 21.
    Phanish MK, Winn SK, Dockrell ME (2009) Connective tissue growth factor-(CTGF, CCN2): a marker, mediator and therapeutic target for renal fibrosis. Nephron Exp Nephrol 114:e83–e92PubMedCrossRefGoogle Scholar
  22. 22.
    Tamm ER, Toris CB, Crowston JG, Sit A, Lim S, Lambrou G, Alm A (2007) In: Weinreb RN, Brandt JD, Garway-Heath D, Medeiros F (eds) Basic science of intraocular pressure. Kugler Publications, Amsterdam, pp 1–14Google Scholar
  23. 23.
    Shepard AR, Millar JC, Pang IH, Jacobson N, Wang WH, Clark AF (2010) Adenoviral gene transfer of active human transforming growth factor-2 elevates intraocular pressure and reduces outflow facility in rodent eyes. Invest Ophthalmol Vis Sci 51(4):2067–2076PubMedCrossRefGoogle Scholar
  24. 24.
    Tamm ER (2009) The trabecular meshwork outflow pathways: structural and functional aspects. Exp Eye Res 88:648–655PubMedCrossRefGoogle Scholar
  25. 25.
    Zenkel M, Krysta A, Pasutto F, Juenemann A, Kruse FE, Schlötzer-Schrehardt U (2011) Regulation of lysyl oxidase-like 1 (LOXL1) and elastin-related genes by pathogenic factors associated with pseudoexfoliation syndrome. Invest Ophthalmol Vis Sci 52(11):8488–8495PubMedCrossRefGoogle Scholar
  26. 26.
    Zhaozeng Lu, Zhang Yuyan, Freddo Thomas F, Gong Haiyan (2011) Similar hydrodynamic and morphological changes in the aqueous humor outflow pathway after washout and Y27632 treatment in monkey eyes. Exp Eye Res 93(4):397–404CrossRefGoogle Scholar
  27. 27.
    Bollinger KE, Crabb JS, Yuan X, Putliwala T, Clark AF, Crabb JW (2011) Quantitative proteomics: TGFβ signaling in trabecular meshwork cells. Invest Ophthalmol Vis Sci 52(11):8287–8294PubMedCrossRefGoogle Scholar
  28. 28.
    Iyer P, Maddala R, Pattabiraman PP, Rao PV (2012) Connective tissue growth factor-mediated upregulation of neuromedin U expression in trabecular meshwork cells and its role in homeostasis of aqueous humor outflow. Invest Ophthalmol Vis Sci 53(8):4952–4962Google Scholar
  29. 29.
    Junglas B, Kuespert S, Seleem AA, Struller T, Ullmann S, Bösl M, Bosserhoff A, Köstler J, Wagner R, Tamm ER, Fuchshofer R (2012) Connective tissue growth factor causes glaucoma by modifying the actin cytoskeleton of the trabecular meshwork. Am J Pathol 180(6):2386–2403Google Scholar
  30. 30.
    Robertson JV, Golesic E, Gauldie J, West-Mays JA (2010) Ocular gene transfer of active TGF-β induces changes in anterior segment morphology and elevated IOP in rats. Invest Ophthalmol Vis Sci 51(1):308–318PubMedCrossRefGoogle Scholar
  31. 31.
    Ghanem AA, Arafa LF, El-Baz A (2011) Connective tissue growth factor and tissue inhibitor of matrix metalloproteinase-2 in patients with exfoliative glaucoma. Curr Eye Res 36(6):540–545PubMedCrossRefGoogle Scholar
  32. 32.
    Browne JG, Ho SL, Kane R, Oliver N, Clark AF, O’Brien CJ, Crean JK (2011) Connective tissue growth factor is increased in pseudoexfoliation glaucoma. Invest Ophthalmol Vis Sci 52(6):3660–3666PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Ying Su
    • 1
    • 2
  • Jingli Cheng
    • 1
  • Hongtao Liu
    • 1
  • Feng Wang
    • 1
  • Shiguang Zhao
    • 3
  1. 1.Department of OphthalmologyFirst Clinical College of Harbin Medical UniversityHarbinChina
  2. 2.Department of Ophthalmology, Department of Anatomy and NeurobiologyBoston University School of MedicineBostonUSA
  3. 3.Department of NeurosurgeryFirst Clinical College of Harbin Medical UniversityHarbinChina

Personalised recommendations