Adenoviral p53 gene transfer inhibits human Tenon's capsule fibroblast proliferation

Johnson, Kristian; Rödicker, Florian; Heise, Kerstin; Heinz, Carsten; Steuhl, Klaus‐Peter; Pützer, Brigitte M.; Hudde, Tobias

doi:10.1136/bjo.2004.051664

Cited by 15 publications

(10 citation statements)

References 24 publications

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“…Both subconjunctival injection and topical treatment can deliver recombinant adenovirus containing the p21 gene (rAd-p21) to conjunctiva to prevent cell proliferation after glaucoma surgery [15,22] . The vector rAd-p53 induced a marked overexpression of p53 in human Tenon's fibroblasts, leading to increased expression of p21 CIP1 , which regulated wound healing following glaucoma surgery [16] . Other experiments have shown efficient expression of reporter genes (CAT and β-gal) in glaucoma surgery tissues (conjunctival fibroblasts) using plasmid or adenovirus constructs injected postoperatively into the filtering bleb [23] .…”

Section: Discussionmentioning

confidence: 99%

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Adenovirus-mediated delivery of p27KIP1 to prevent wound healing after experimental glaucoma filtration surgery

et al. 2009

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Aim:The aim of the study was to evaluate the outcome of adenovirus-mediated p27 KIP1 (Ad-p27) expression on wound healing after filtration surgery and to investigate the inhibition of cell proliferation induced by Ad-p27. Methods: We constructed the adenovirus recombinant vector Ad-p27 and administered it to a rabbit model of glaucoma filtration surgery by subconjunctival injection; phosphate-buffered saline (PBS) and mitomycin C (MMC) were used as controls. Intraocular pressure (IOP), bleb scores, and anterior chamber depths were observed during a 28-d period. Histological examinations, fluorescence observations and Western blot analyses were evaluated. Results: Ad-p27 enhanced the surgical outcome and inhibited cell proliferation when compared with PBS. Bleb scores in the Ad-p27-treated eyes were higher than those in the PBS-treated eyes on d 7 (P<0.01), 14 (P<0.01) and 21 (P<0.05). On d 28, IOP remained significantly decreased in the Ad-p27 group compared with the PBS group (P<0.05). However, no differences in bleb scores or IOPs were observed between the Ad-p27 and MMC groups. Histological analysis showed that total cell numbers were markedly reduced, and less scar tissue was observed at the surgical site in eyes treated with Ad-p27. The number of fibroblasts was decreased in Tenon's capsule in Ad-p27-treated eyes; however, a marked and diffuse signal from the green fluorescent protein (GFP) was observed in fibroblasts. Western blot analysis revealed a high level of p27 KIP1 expression in conjunctival epithelium (P<0.01), relatively high expression in superficial scleral stroma (P<0.01), and low expression in corneal epithelium in the Ad-p27 group. Conclusions: Ad-p27 administration significantly improves the outcome of filtration surgery and inhibits postoperative proliferation in rabbit eyes. These findings suggest that p27 KIP1 is a potential adjunctive agent for inhibition of wound healing after filtration surgery.

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Section: Discussionmentioning

confidence: 99%

“…Gene delivery to Tenon's capsule fibroblasts by viral vectors is effective in inhibiting cell proliferation in several models of filtration surgery [15,16] . The hypothesis of our study was that a recombinant p27 KIP1 would prevent wound healing in experimental filtration surgery.…”

Section: Introductionmentioning

confidence: 99%

Adenovirus-mediated delivery of p27KIP1 to prevent wound healing after experimental glaucoma filtration surgery

et al. 2009

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“…The cytotoxicity was significant when MOI reached above 500, and there was no influence on cell growth when MOI was below 50 (36)(37)(38). In present research, relatively stable infecting efficiency (>95%) could be obtained when MOI of rAd was in the range of 100 to 500, and no significant difference was detected in proliferative inhibition ratio of POE-9n cell in MTT experiment, whereas in the MOI = 500 group, significant cytotoxic effect was observed.…”

Section: Discussionmentioning

confidence: 99%

In vitro and Clinical Studies of Gene Therapy with Recombinant Human Adenovirus-p53 Injection for Oral Leukoplakia

Zhang

et al. 2009

Clinical Cancer Research

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Purpose: Oral leukoplakia is a well-recognized precancerous lesion of squamous cell carcinoma. When accompanied with abnormal p53 expression, it suffered a higher risk of canceration. The present study was carried out to test whether the recombinant human adenovirus-p53 could introduce wild-type p53 gene to oral leukoplakia cells and induce cell cycle arrest and apoptosis. Experimental Design: We select p53(-) oral dysplastic keratinocyte POE-9n, to observe the growth inhibition, cell cycle change, apoptosis-induced effects, and elaborate the corresponding molecular mechanism of recombinant adenovirus-p53 on POE-9n cells. Meanwhile, we evaluate the feasibility, safety, and biological activity of multipoints intraepithelial injections of recombinant adenovirus-p53 in 22 patients with dysplastic oral leukoplakia. Results: Exogenous p53 could be successfully transduced into POE-9n cells by recombinant adenovirus-p53. The optimal infecting titer in this study was multiplicity of infection (MOI) = 100. Recombinant adenovirus-p53 could strongly inhibit cell proliferation, induce apoptosis, and arrest cell cycle in stage G 1 in POE-9n cells by inducing p21 CIP/WAF and downregulating bcl-2 expression. In the posttreatment patients, p53 protein and p21 CIP/WAF protein expression were significantly enhanced, yet bcl-2 protein presented low expression. Sixteen patients showed clinical response to the treatment, and 14 patients showed obvious histopathologic improvement. Conclusion: Intraepithelial injections of recombinant human adenovirus-p53 were safe, feasible, and biologically active for patients with dysplastic oral leukoplakia. (Clin Cancer Res 2009;15(21):6724-31)

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“…Thus, inhibiting the proliferation of Tenon's fibroblasts during the wound healing is an important antiscarring strategy. [1][2][3][4]17,18 The importance of CDKN1B in regulating cell proliferation has been demonstrated in varied animal models. It has been shown that there was markedly decreased expression of CDKN1B in the corneal endothelium cells after injury in a rat model of connexin43 knockdown, 31 whereas the adenovirusmediated overexpression of CDKN1B inhibited vascular smooth muscle cells proliferation, 32 and led to cell growth arrest in human renal carcinoma cells.…”

Section: Discussionmentioning

confidence: 99%

“…However, excessive scarring of the conjunctiva at the bleb and sclerostomy sites after GFS often leads to the failure of the surgery. [1][2][3][4] It has been proved that the proliferation of fibroblasts from the subconjunctival space is the major cause of the scarring process. 5,6 Both positive and negative regulators of cell-cycle activity are the critical regulators of cell proliferation and differentiation.…”

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confidence: 99%

Overexpression of CDKN1B Inhibits Fibroblast Proliferation in a Rabbit Model of Experimental Glaucoma Filtration Surgery

Yang

Deng

Zhou

et al. 2013

Invest. Ophthalmol. Vis. Sci.

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PURPOSE. To investigate the potential antiproliferative effect of cyclin-dependent kinase inhibitor 1B (CDKN1B) overexpression in a rabbit model of glaucoma filtration surgery (GFS).METHODS. The recombinant adenovector expressing exogenous CDKN1B was delivered to Tenon's capsule by subconjunctival injection during unilateral filtration surgery. The time course of CDKN1B expression was monitored by immunohistochemistry and Western blot analysis. Evaluation of proliferating activity was performed by proliferating cell nuclear antigen (PCNA), argyrophilic nucleolar organizing region (AgNOR) staining, and fibroblast-specific protein 1 (FSP-1). Cyclin-dependent kinase 2 (Cdk2) and Cdk4 expression were detected with immunohistochemical analysis.RESULTS. The overexpression of CDKN1B in Tenon's capsule was monitored throughout the experimental period. Immunoreactivity to CDKN1B was mainly observed in the nucleus of fibroblasts. The increased expression of CDKN1B in sclera was detected up to 21 days after viral infection, whereas the level of CDKN1B protein in corneal stroma was not significantly increased. The overexpression of CDKN1B induced a significant decrease in AgNOR number/nucleus and area/nucleus, PCNA staining, FSP-1 positive cells, and the decreased expressions of Cdk2 and Cdk4, as evidenced by nuclear and cytoplasmic immunoreactivity to Cdk2 and Cdk4 antibodies in positive fibroblasts.CONCLUSIONS. The persistent overexpression of CDKN1B mediated by the recombinant adenovector expressing exogenous CDKN1B in Tenon's fibroblasts after GFS may lead to the inhibition of fibroblast proliferation and the downregulation of Cdk2 and Cdk4 activity, thereby reducing the severity of scar formation and the surgical outcome. (Invest Ophthalmol Vis Sci. 2013;54:343-352)

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Adenoviral p53 gene transfer inhibits human Tenon's capsule fibroblast proliferation

Cited by 15 publications

References 24 publications

Adenovirus-mediated delivery of p27KIP1 to prevent wound healing after experimental glaucoma filtration surgery

Adenovirus-mediated delivery of p27KIP1 to prevent wound healing after experimental glaucoma filtration surgery

In vitro and Clinical Studies of Gene Therapy with Recombinant Human Adenovirus-p53 Injection for Oral Leukoplakia

Overexpression of CDKN1B Inhibits Fibroblast Proliferation in a Rabbit Model of Experimental Glaucoma Filtration Surgery

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