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AAV2/1-TNFR:Fc gene delivery prevents periodontal disease progression.

Cirelli JA, Park CH, MacKool K, Taba M, Lustig KH, Burstein H, Giannobile WV - Gene Ther. (2008)

Bottom Line: Therapeutic control of progressive tissue destruction in high-risk patients is a significant challenge in therapy.AAV2/1-TNFR:Fc therapy led to sustained therapeutic levels of serum TNFR protein and protected against Pg-LPS-mediated loss of bone volume and density.Furthermore, AAV2/1-TNFR:Fc administration reduced local levels of multiple proinflammatory cytokines and osteoclast-like cells at the periodontal lesions.

View Article: PubMed Central - PubMed

Affiliation: Department of Periodontics and Oral Medicine, School of Dentistry, University of Michigan, Ann Arbor, MI 48109-1078, USA.

ABSTRACT
Periodontal disease is a chronic inflammatory condition induced by tooth-associated microbial biofilms that induce a host immune response. Therapeutic control of progressive tissue destruction in high-risk patients is a significant challenge in therapy. Soluble protein delivery of antagonists to tumor necrosis factor-alpha (TNF-alpha) inhibits alveolar bone resorption due to periodontitis. However, protein therapy raises several concerns, such as recurrence of disease activity after treatment cessation and repeated dosing regimens. In this study, we used pseudotyped adeno-associated virus vector based on serotype 1 (AAV2/1) to deliver the TNF receptor-immunoglobulin Fc (TNFR:Fc) fusion gene to rats subjected to experimental Porphyromonas gingivalis (Pg)-lipopolysaccharide (LPS)-mediated bone loss. Animals received Pg-LPS delivered to the gingivae thrice weekly for 8 weeks, vehicle alone, Pg-LPS and intramuscular delivery of pseudotyped AAV2/1-TNFR:Fc vector (1 x 10(11) DNase I-resistant particles) or AAV2/1-TNFR:Fc vector delivered to naive animals. AAV2/1-TNFR:Fc therapy led to sustained therapeutic levels of serum TNFR protein and protected against Pg-LPS-mediated loss of bone volume and density. Furthermore, AAV2/1-TNFR:Fc administration reduced local levels of multiple proinflammatory cytokines and osteoclast-like cells at the periodontal lesions. These findings suggest that delivery of AAV2/1-TNFR:Fc may be a viable approach to modulate periodontal disease progression.

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Histological sections (H/E, 200X) of subepithelial (upper panels) and bone crest (lower panels) levels of periodontal tissues around the maxillary second molar of Vehicle (a and d), Pg-LPS (b and e) and AAV2/1-TNFR:Fc + Pg-LPS (c and f) treated animals, at 4 weeks. The number of inflammatory cells was determined by stereometric analysis using a point-counting technique. A 500 μm2 rectangular-lattice grid with 50 intersection points was constructed and the type of structure found on the intersection of the grid lines was counted on a optical microscope. (One-way ANOVA and Tukey’s post hoc tests).
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Figure 1: Histological sections (H/E, 200X) of subepithelial (upper panels) and bone crest (lower panels) levels of periodontal tissues around the maxillary second molar of Vehicle (a and d), Pg-LPS (b and e) and AAV2/1-TNFR:Fc + Pg-LPS (c and f) treated animals, at 4 weeks. The number of inflammatory cells was determined by stereometric analysis using a point-counting technique. A 500 μm2 rectangular-lattice grid with 50 intersection points was constructed and the type of structure found on the intersection of the grid lines was counted on a optical microscope. (One-way ANOVA and Tukey’s post hoc tests).

Mentions: Animals were sacrificed at four and eight weeks after Pg-LPS disease induction. Stereometric histological analysis using hematoxylin and eosin (H/E) staining was performed to evaluate the inflammatory cell infiltrate induced by an 8-week course of local Pg-LPS administration with or without IM AAV2/1-TNFR:Fc gene therapy. At both timepoints, an intense inflammatory cell infiltrate was consistently observed in the subepithelial connective tissue and surrounding alveolar bone of the periodontia of Pg-LPS-treated animals (Fig. 1b,e). In contrast, a significantly less intense inflammatory reaction was observed in the AAV2/1-TNFR:Fc + Pg-LPS-treated animal (Fig. 1c, f). Control animals (Veh, NT and vector only) did not display evidence of significant inflammatory cell infiltrates (Fig. 1a, d). Statistical analysis demonstrated higher number of inflammatory cells for diseased group (Pg-LPS) in both analyzed areas at 4 weeks timepoint and in subepithelial area at 8 weeks timepoint when compared to treated animals (AAV2/1-TNFR:Fc + Pg-LPS) (Fig. 1). These results demonstrate the anti-inflammatory effect of TNFR:Fc in the face of continued Pg-LPS challenge.


AAV2/1-TNFR:Fc gene delivery prevents periodontal disease progression.

Cirelli JA, Park CH, MacKool K, Taba M, Lustig KH, Burstein H, Giannobile WV - Gene Ther. (2008)

Histological sections (H/E, 200X) of subepithelial (upper panels) and bone crest (lower panels) levels of periodontal tissues around the maxillary second molar of Vehicle (a and d), Pg-LPS (b and e) and AAV2/1-TNFR:Fc + Pg-LPS (c and f) treated animals, at 4 weeks. The number of inflammatory cells was determined by stereometric analysis using a point-counting technique. A 500 μm2 rectangular-lattice grid with 50 intersection points was constructed and the type of structure found on the intersection of the grid lines was counted on a optical microscope. (One-way ANOVA and Tukey’s post hoc tests).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2656404&req=5

Figure 1: Histological sections (H/E, 200X) of subepithelial (upper panels) and bone crest (lower panels) levels of periodontal tissues around the maxillary second molar of Vehicle (a and d), Pg-LPS (b and e) and AAV2/1-TNFR:Fc + Pg-LPS (c and f) treated animals, at 4 weeks. The number of inflammatory cells was determined by stereometric analysis using a point-counting technique. A 500 μm2 rectangular-lattice grid with 50 intersection points was constructed and the type of structure found on the intersection of the grid lines was counted on a optical microscope. (One-way ANOVA and Tukey’s post hoc tests).
Mentions: Animals were sacrificed at four and eight weeks after Pg-LPS disease induction. Stereometric histological analysis using hematoxylin and eosin (H/E) staining was performed to evaluate the inflammatory cell infiltrate induced by an 8-week course of local Pg-LPS administration with or without IM AAV2/1-TNFR:Fc gene therapy. At both timepoints, an intense inflammatory cell infiltrate was consistently observed in the subepithelial connective tissue and surrounding alveolar bone of the periodontia of Pg-LPS-treated animals (Fig. 1b,e). In contrast, a significantly less intense inflammatory reaction was observed in the AAV2/1-TNFR:Fc + Pg-LPS-treated animal (Fig. 1c, f). Control animals (Veh, NT and vector only) did not display evidence of significant inflammatory cell infiltrates (Fig. 1a, d). Statistical analysis demonstrated higher number of inflammatory cells for diseased group (Pg-LPS) in both analyzed areas at 4 weeks timepoint and in subepithelial area at 8 weeks timepoint when compared to treated animals (AAV2/1-TNFR:Fc + Pg-LPS) (Fig. 1). These results demonstrate the anti-inflammatory effect of TNFR:Fc in the face of continued Pg-LPS challenge.

Bottom Line: Therapeutic control of progressive tissue destruction in high-risk patients is a significant challenge in therapy.AAV2/1-TNFR:Fc therapy led to sustained therapeutic levels of serum TNFR protein and protected against Pg-LPS-mediated loss of bone volume and density.Furthermore, AAV2/1-TNFR:Fc administration reduced local levels of multiple proinflammatory cytokines and osteoclast-like cells at the periodontal lesions.

View Article: PubMed Central - PubMed

Affiliation: Department of Periodontics and Oral Medicine, School of Dentistry, University of Michigan, Ann Arbor, MI 48109-1078, USA.

ABSTRACT
Periodontal disease is a chronic inflammatory condition induced by tooth-associated microbial biofilms that induce a host immune response. Therapeutic control of progressive tissue destruction in high-risk patients is a significant challenge in therapy. Soluble protein delivery of antagonists to tumor necrosis factor-alpha (TNF-alpha) inhibits alveolar bone resorption due to periodontitis. However, protein therapy raises several concerns, such as recurrence of disease activity after treatment cessation and repeated dosing regimens. In this study, we used pseudotyped adeno-associated virus vector based on serotype 1 (AAV2/1) to deliver the TNF receptor-immunoglobulin Fc (TNFR:Fc) fusion gene to rats subjected to experimental Porphyromonas gingivalis (Pg)-lipopolysaccharide (LPS)-mediated bone loss. Animals received Pg-LPS delivered to the gingivae thrice weekly for 8 weeks, vehicle alone, Pg-LPS and intramuscular delivery of pseudotyped AAV2/1-TNFR:Fc vector (1 x 10(11) DNase I-resistant particles) or AAV2/1-TNFR:Fc vector delivered to naive animals. AAV2/1-TNFR:Fc therapy led to sustained therapeutic levels of serum TNFR protein and protected against Pg-LPS-mediated loss of bone volume and density. Furthermore, AAV2/1-TNFR:Fc administration reduced local levels of multiple proinflammatory cytokines and osteoclast-like cells at the periodontal lesions. These findings suggest that delivery of AAV2/1-TNFR:Fc may be a viable approach to modulate periodontal disease progression.

Show MeSH
Related in: MedlinePlus