PMC:2575619 / 31108-36611 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/2575619","sourcedb":"PMC","sourceid":"2575619","source_url":"https://www.ncbi.nlm.nih.gov/pmc/2575619","text":"Evaluation of alternative splice variant activity in an in vivo model of arthritis\nSince angiogenesis plays a key role in RA, we next evaluated the therapeutic potential of ASV in an extensively validated mouse model of arthritis – namely, acute CIA. On the day of disease onset, replication-incompetent alternative splice variant-expressing adenoviruses were administered as a single dose of 1 × 107 plaque-forming units. The severity of arthritis in the mice was consecutively recorded for the following 10 days.\nControl adenovirus (LacZ) was without significant effect on disease severity (Table 3 and Figures 5 and 6). In contrast, treatment with either Tie1-751 (Table 3 and Figure 5) or VEGFR1-541 (Table 3 and Figure 6) alternative splice variant adenoviruses significantly reduced disease severity, as evidenced by decreased clinical scores (P \u003c 0.001), reduced paw thickness (P \u003c 0.001), and reduced joint inflammation and destruction (P \u003c 0.01 and P \u003c 0.001 for VEGFR1-541 and Tie1-751, respectively). An example of the joint histology for untreated, LacZ ASV-treated and Tie1-751 ASV-treated mice is shown in Figure 5c, with quantitative analysis of the histology depicted in Table 3.\nTable 3 Effect of alternative splice variant-expressing adenoviruses on joint inflammation and destruction Following onset of arthritis, mice were treated with the alternative splice variant adenovirus indicated. Data presented as P values of mice treated with the indicated recombinant alternative splice variant adenoviruses as compared with untreated mice, and are expressed as the P value of clinical scores, paw swelling, and histological evaluation. For clinical scores and paw swelling, data were analyzed using two-way analysis of variance versus untreated mice. For histological evaluation, H \u0026 E and toluidine blue stained sections were scored for pannus formation, synovitis, and bone and cartilage erosion. Data were analyzed using the chi-square test for trend versus untreated mice.\nFigure 5 Inhibition of murine collagen-induced arthritis by Tie1-751. On the day of arthritis onset, mice received intravenously 1 × 107 plaque-forming units of adenoviruses expressing either LacZ (○) or Tie1-751 alternative splice variants (ASV) (●), or remained untreated (□) as indicated. (a) Clinical score was recorded daily, and data were analyzed by two-way analysis of variance versus untreated mice. LacZ, not significant (P = 0.3734); Tie1-751, P \u003c 0.001; n = 6 per group. (b) Paw swelling was recorded using calipers daily, and data were analyzed by two-way analysis of variance versus untreated mice. LacZ, not significant (P = 0.5134); Tie1-751, P \u003c 0.001. Data are means of n = 6. (c) Serial sections of mouse hind feet were stained with either H \u0026 E (left panels) or toluidine blue (right panels). Figure shows tibia–tarsus joint sections from untreated mice (top panels), from LacZ adenovirus-treated mice (middle panels), and from Tie1-751 ASV adenovirus-treated mice (bottom panels). Sections are shown at 40× magnification; scale bar = 20 μm. (d) Pharmacokinetics of Tie1-751 from the ASV-expressing adenovirus. Sera from untreated mice or mice treated intravenously with 1 × 109 plaque-forming units of Tie1-751 ASV adenovirus were analyzed after the indicated times by western blot, followed by scanning and quantitation using Tie1-751 standard. (e) Effect of recombinant Tie1-751-Fc protein on clinical score. Results are from mice on day 10 of arthritis. Filled bars, untreated mice; empty bars, mice treated with recombinant Tie1-751-Fc 30 mg/kg, three times weekly. Data are means of n = 6. **P \u003c 0.01 for Tie-751-Fc treated mice versus untreated mice.\nFigure 6 Differential effects of alternative splice variant-expressing adenoviruses on collagen-induced arthritis mice. On the day of arthritis onset, mice received intravenously 1 × 107 plaque-forming units of the indicated alternative splice variant (ASV)-expressing adenoviruses. Clinical scores ((a), (c), and (e))and paw thickness measured by calipers ((b), (d), and (f))were recorded daily. Data were analyzed by two-way analysis of variance versus untreated mice (Table 3). (a) and (b) Mice received adenoviruses expressing either LacZ (○), VEGFR1-541 (■), VEGFR2-712 (▲) or VEGFR3-765 (●), or remained untreated (□). Data are means of n = 5 per group. (c) and (d) Mice received adenoviruses expressing either LacZ (○), Met-877 (■), Tie1-751 (▲) or FGFR1-320 (●), or remained untreated (□). Data are means of n = 6 per group. (e) and (f) Mice received adenoviruses expressing either LacZ (○), RAGE-387 (■), PDGFRβ-366 (▲), c-Kit-413 (●), or CSF1R-306 (◆), or remained untreated (□). Data are means of n = 6 per group. The presence of Tie1-751 in mouse sera was confirmed by western blotting (Figure 5d). The effectiveness of Tie1-751 in CIA was confirmed using recombinant Tie1-751-Fc protein (Figure 5e).\nA less marked disease-modifying effect was seen with the adenovirus encoding FGFR1-320 (Table 3 and Figure 6), which reduced clinical scores and paw thickness (P \u003c 0.01) but without achieving a statistically significant improvement of joint histological evaluation (P \u003c 0.057). Similarly, VEGFR2-712 reduced the clinical score (P \u003c 0.001) but failed to affect the paw thickness and the histological scores (Table 3 and Figure 6).\nTreatment with ASV derived from VEGFR3, RAGE, Met, c-Kit, PDGFRβ, and CSF1R adenoviruses did not generate a significant effect on any of the disease parameters (Table 3 and Figure 6).","divisions":[{"label":"Title","span":{"begin":0,"end":82}},{"label":"Table caption","span":{"begin":1196,"end":1995}},{"label":"Figure caption","span":{"begin":1995,"end":3675}},{"label":"Figure caption","span":{"begin":3674,"end":4701}}],"tracks":[]}