Proteinase and Growth Factor Alterations Revealed by Gene Microarray Analysis of Human Diabetic Corneas

Saghizadeh, Mehrnoosh; Kramerov, Andrei A.; Tajbakhsh, Jian; Aoki, Annette M.; Wang, Charles; Chai, Ningli; Ljubimova, Julia Y.; Sasaki, Takako; Sosne, Gabriel; Carlson, Marc; Nelson, Stanley F.; Ljubimov, Alexander V.

doi:10.1167/iovs.04-1507

Cited by 76 publications

(88 citation statements)

References 73 publications

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“…[42][43][44][45] Cathepsins B, G, and F have been implicated in fibrosis at Bowman layer breaks of keratoconus corneas and in corneal wound healing. 23,46,47 These data suggest the occurrence of ongoing proteolysis around the INTACS channels, possibly as an attempt of corneal stromal cells to remodel the stroma and remove iatrogenic accumulations of fibrotic ECM. Future studies of this process may focus on cytokines that control ECM accumulation and proteinase activation.…”

Section: Discussionmentioning

confidence: 83%

“…MMP-3 and MMP-10 (stromelysin-1 and -2, respectively), as well as MMP-7, which can degrade ECM and BM components, have been implicated in corneal wound healing and stromal remodeling. 23,40,41 Although cathepsins are largely considered lysosomal proteinases, some of them may be secreted (cathepsins B, H, K) or have extracellular localization, such as cathepsin V/L2. [42][43][44][45] Cathepsins B, G, and F have been implicated in fibrosis at Bowman layer breaks of keratoconus corneas and in corneal wound healing.…”

Section: Discussionmentioning

confidence: 99%

“…20 Routine specificity controls without primary or secondary antibodies were negative. Well-characterized antibodies [20][21][22][23][24] were used for α1/α2, α3, α4, and α6 chains of type IV collagen; α2, α4, α5, β1, β2, and γ1 chains of laminin; laminin-332; fibronectin 8th type III repeat and cellular (ED-A repeat containing) fibronectin; nidogen-1; nidogen-2; types III, VI, VIII, and XIV collagen; perlecan; tenascin-C; vitronectin; thrombospondin-1; fibrillin-1; α-smooth-muscle actin; matrix metalloproteinases (MMP)-1, -2, -3, -7, -9, and -10; urokinase; and cathepsins B, F, H, and L. Monoclonal antibody III-53 to type III collagen was from EMD Biosciences/Calbiochem (La Jolla, CA), monoclonal antibody HIM6 to EMMPRIN/CD147/neurothelin was from BD Biosciences Pharmingen (San Jose, CA), and polyclonal antibody to MMP-10 hinge region was from AnaSpec (San Jose, CA).…”

Section: Immunohistochemistrymentioning

confidence: 99%

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Alterations of Extracellular Matrix Components and Proteinases in Human Corneal Buttons With INTACS for Post-Laser In Situ Keratomileusis Keratectasia and Keratoconus

Maguen¹,

Rabinowitz²,

Regev³

et al. 2008

Cornea

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Purpose-To perform an immunohistochemical evaluation of corneas with INTACS for post-laser in situ keratomileusis (LASIK) keratectasia and keratoconus, obtained after corneal transplantation.Methods-Corneas from 1 patient with INTACS for post-LASIK keratectasia and 2 patients with INTACS for keratoconus were obtained within 3 hours after penetrating keratoplasty, and cryostat sections were analyzed by immunostaining for 35 extracellular matrix (ECM) components and proteinases.Results-In the stroma of all corneas next to an INTACS implant, ECM components typically associated with fibrosis were observed. These included tenascin-C, fibrillin-1, and types III, IV (α1/ α2 chains), and XIV collagen. Also, significant deposition of perlecan, nidogen-2, and cellular fibronectin was revealed in the same locations. The keratoconus cases displayed typical Bowman layer breaks and subepithelial fibrosis with deposition of various ECM components. In all cases, some keratocytes around INTACS were positive for specific proteinases associated with stromal remodeling, including cathepsins F and H, matrix metalloproteinase (MMP)-1, MMP-3, and MMP-10. Staining for MMP-7 was variable; MMP-2 and MMP-9 were mostly negative. Patterns of type IV collagen α3, α4, and α6 chains; types VI and VIII collagen;α4, α5,β1, β2, and γ1 laminin chains; vitronectin; thrombospondin-1; urokinase; EMMPRIN; and cathepsins B and L were unchanged around INTACS in all 3 cases compared with normal. Recently, INTACS has acquired new interest as an alternative treatment of keratoconus. [5][6][7][8][9][10] The rationale is that INTACS not only flattens the central cornea but also corrects some of the astigmatic component. The corneal profile becomes more symmetrical, usually allowing better correction with glasses and contact lenses. Given that, for these patients, the alternative would be penetrating keratoplasty, INTACS may delay the need for this option. 10,11 Midstromal semicircular channels for INTACS ring insertion are created by a mechanical system with a blunt spiral device. 1,12,13 Despite the streamlined mechanistic procedure, attempts were made to improve control over the depth and shape of the channels. A femtosecond laser mounted on a delivery system capable of creating an intrastromal pattern of virtually any shape was recently used to create channels for INTACS insertion. 10 This method may be technically easier and more precise, reflecting better final outcomes. Conclusions-AbnormalKeratectasia is a rare but serious complication of laser vision correction of any refractive errors. It occurs mostly after LASIK but was also described after surface excimer laser ablation (photorefractive keratectomy). 14-16 It involves severe refractive changes mostly occurring several years after the surgery, which are caused by thinning and sagging of the corneal profile, similar to keratoconus.The etiology of this complication remains uncertain. In some cases, preexisting forme fruste keratoconus was documented. 17,18 Other cases could be caused by a me...

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Section: Discussionmentioning

confidence: 83%

Section: Discussionmentioning

confidence: 99%

Section: Immunohistochemistrymentioning

confidence: 99%

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Alterations of Extracellular Matrix Components and Proteinases in Human Corneal Buttons With INTACS for Post-Laser In Situ Keratomileusis Keratectasia and Keratoconus

Maguen¹,

Rabinowitz²,

Regev³

et al. 2008

Cornea

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“…Examples of molecules down-regulated to undetectable levels upon this phenotypic change include the keratin sulfate proteoglycans, lumican and keratocan, and the corneal crystallins, aldehyde dehydrogenase-1, aldehyde dehydrogenase-3 and α-transketolase (Saghizadeh et al, 2005). CpG islands in the 5′ UTR and 1000 bp upstream of the start site were found in aldehyde dehydrogenase-3 and α-transketolase using MethPrimer.…”

Section: Discussionmentioning

confidence: 99%

Epigenetic silencing of maspin expression occurs early in the conversion of keratocytes to fibroblasts

Horswill

Narayan

Warejcka

et al. 2008

Experimental Eye Research

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Maspin, a 42 kDa non-classical serpin (serine protease inhibitor) that controls cell migration and invasion, is mainly expressed by epithelial-derived cells but is also expressed in corneal stromal keratocytes. Upon culture of stromal keratocytes in the presence of FBS, maspin is down regulated to nearly undetectable levels by passage two. DNA methylation is one of several processes that controls gene expression during cell differentiation, development, genetic imprinting, and carcinogenesis but has not been studied in corneal stromal cells. The purpose of this study was to determine whether DNA methylation of the maspin promoter and histone H3 dimethylation are involved in the mechanism of down regulation of maspin synthesis in human corneal stromal fibroblasts and myofibroblasts. Human donor corneal stroma cells were immediately placed into serum-free defined medium or cultured in the presence of FBS and passed into serum-free medium or medium containing FBS or FGF-2 to induce the fibroblast phenotype or TGF-β1 for the myofibroblast phenotype. These cell types are found in wounded corneas. The cells were used to prepare RNA for semi-quantitative or quantitative RT-PCR or to extract protein for western analysis. In addition, P4 FBS cultured fibroblasts were treated with the DNA demethylating agent, 5-aza-2′-deoxycytidine (5-Aza-dC), and the histone deacetylase inhibitor, trichostatin A (TSA). Cells with and without treatment were harvested and assayed for DNA methylation using sodium bisulfite sequencing. The methylation state of histone H3 associated with the maspin gene in the P4 fibroblast cells was determined using a ChIP assay. Freshly harvested corneal stromal cells expressed maspin but upon phenotypic differentiation, maspin mRNA and protein were dramatically down-regulated. Sodium bisulfite sequencing revealed that the maspin promoter in the freshly isolated stromal keratocytes was hypomethylated while both the P0 stromal cells and the P1 cells cultured in the presence of serum free defined medium, FGF-2 and TGF-β1 were hypermethylated. Down regulation of maspin synthesis was also associated with histone H3 dimethylation at Lysine 9. Both maspin mRNA and protein were reexpressed at low levels with 5-Aza-dC but not TSA treatment. Addition of TSA to 5-Aza-dC treated cells did not increase maspin expression. Treatment with 5-Aza-dC did not significantly alter demethylation of the maspin promoter but did demethylate histone H3. These results show maspin promoter hypermethylation and histone methylation occur with down regulation *Corresponding Author: Sally S. Twining, PhD, Department of Biochemistry, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, Phone: 414-456-8431, Fax: 414-456-6510, e-mail: stwining@mcw.edu. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of...

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“…The PCR for ANG II receptor subtypes AT1 and AT2 was performed using specific pairs of primers previously described by Hou et al (32) and carried out with the following profile: 2 min at 95°C for 1 cycle, 1 min at 95°C and 1 min at 60°C for 35 cycles, and 7 min at 72°C. For the amplification of the human collagen type IV (chain ␣ 1), collagen type I (chain ␣1), and laminin (chain ␣4), primers previously described were used (13,55,59). Collagen type IV (chain ␣1) gene was amplified by a first step of 15 s at 95°C, followed by 1 min at 60°C for 40 cycles, and 7 min at 72°C.…”

Section: Compounds and Drugsmentioning

confidence: 99%

Regulation of angiotensin II receptors and extracellular matrix turnover in human retinal pigment epithelium: role of angiotensin II

Striker

Praddaude

Garnica

et al. 2008

American Journal of Physiology-Cell Physiology

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Striker GE, Praddaude F, Alcazar O, Cousins SW, MarinCastañ o ME. Regulation of angiotensin II receptors and extracellular matrix turnover in human retinal pigment epithelium: role of angiotensin II. Am J Physiol Cell Physiol 295: C1633-C1646, 2008. First published October 15, 2008 doi:10.1152/ajpcell.00092.2008.-The early stage of age-related macular degeneration (AMD) is characterized by the formation of subretinal pigment epithelium (RPE) deposits as a result of the dysregulation in the turnover of extracellular matrix (ECM) molecules. However, the mechanism involved remains unclear. Hypertension (HTN) is an important risk factor for AMD, and angiotensin II (ANG II) is the most important hormone associated with HTN. However, the relevance of ANG II receptors and ANG II effects on RPE have not been investigated yet. Therefore, the expression and regulation of ANG II receptors as well as the ECM turnover were studied in human RPE. ANG II receptors were expressed and upregulated by ANG II in human RPE. This regulation resulted in functional receptor expression, since an increase in intracellular concentration of calcium was observed upon ANG II stimulation. ANG II also increased matrix metalloproteinase (MMP)-2 activity and MMP-14 at the mRNA and protein levels as well as type IV collagen degradation. These ANG II effects were abolished in the presence of the ANG II receptor subtype 1 (AT1) receptor antagonist candesartan. In contrast, ANG II decreased type IV collagen via both AT1 and AT2 receptors, suggesting a synergistic effect of the two receptor subtypes. In conclusion, we have confirmed the presence of ANG II receptors in human RPE and their regulation by ANG II as well as the regulation of ECM molecules via ANG II receptors. Our data support the hypothesis that ANG II may exert biological function in RPE through ANG II receptors and that ANG II may cause dysregulation of molecules that play a major role in the turnover of ECM in RPE basement membrane and Bruch's membrane, suggesting a pathogenic mechanism to explain the link between HTN and AMD. calcium; hypertension; Bruch's membrane EARLY AGE-RELATED MACULAR DEGENERATION (AMD) is one of the most common irreversible causes of severe loss of vision in elderly population (23,39). Dysfunction of retinal photoreceptors is the ultimate cause of vision loss. However, the initial pathogenic target of AMD is the retinal pigment epithelium (RPE) and the subjacent extracellular matrix (Bruch's membrane, or BrM) (25). Although age is the major determinant for developing AMD, clinical studies have revealed a number of systemic and environmental risk factors (23,49,54). Among them, hypertension (HTN) is of special relevance (29,34,37,39). However, the mechanism(s) by which HTN may affect AMD remains unclear. Interestingly, different reports have related that angiotensin II (ANG II), the final product of the renin-angiotensin system (RAS) and the most important hormone associated with HTN, may contribute to chronic diseases (17,43,58,69).Expression and localization ...

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Proteinase and Growth Factor Alterations Revealed by Gene Microarray Analysis of Human Diabetic Corneas

Cited by 76 publications

References 73 publications

Alterations of Extracellular Matrix Components and Proteinases in Human Corneal Buttons With INTACS for Post-Laser In Situ Keratomileusis Keratectasia and Keratoconus

Alterations of Extracellular Matrix Components and Proteinases in Human Corneal Buttons With INTACS for Post-Laser In Situ Keratomileusis Keratectasia and Keratoconus

Epigenetic silencing of maspin expression occurs early in the conversion of keratocytes to fibroblasts

Regulation of angiotensin II receptors and extracellular matrix turnover in human retinal pigment epithelium: role of angiotensin II

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