Angiogenesis in Viral Immunoinflammatory Lesions

1. Angiogenesis in Viral Immunoinflammatory Lesions Barry T. Rouse University of Tennessee Part 4

2. Does control of angiogenesis represent a smart approach to control stromal keratitis? • Problem: many factors to deal with

3. EMAP IIEndothelial Monocyte Activating Polypeptide • Cytokine with several effects on neovascularization • Apoptosis of vascular endothelial cells • Upregulates TNF receptors – so TNFa induces apoptosis • Activates tissue factors and blood clots • Proinflammatory effects • Rationale - targets growing blood vessels so should inhibit effects of several angiogenic factors

4. EMAP II induces apoptosis in vascular endothelial cells in vitro and in vivo 40 * b a Epi * 30 Lim # of Apoptotic Cells/HPF 20 Endo 10 0 0 10 100 1000 EMAP II (ng/ml)

5. EMAP II suppresses neovascularization in murine HSK Control EMAP II 20 c EMAP II i * * c D2 p.I. D8 p.I. D15 p.I. Control * 15 i * c Angiogenesis Score 10 c i * 5 c c 0 0 5 10 15 20 25 Days p.i.

6. Control EMAP II D8 D8 D15 D15 D15 D15 F F E E EMAP II Suppresses HSK lesion score 5 * EMAP II * Control 4 * 3 HSK Lesion Score 2 * 1 0 0 5 10 15 20 25 Days p.i.

7. Future approaches – possible siRNA targeting multiple angiogenesis factors

8. Preliminary study • Plasmid encoding siRNA expressed following intra-stromal or sub-conjunctival injection • Mixture of siRNA inhibiting VEGF and VEGF receptors gave significant inhibition of angiogenesis induced by CpG

9. Inhibition of CpG induced angiogenesis by siRNA delivery CpG + siLacZ Day 4 CpG + siVEGF, R1, R2 Day 4

10. General Conclusions • Herpes induced stromal keratitis an immunoinflammatory lesion • Neovascularization a major step in pathogenesis • HSV induces angiogenesis indirectly • Control of angiogenesis inhibits SK • Need multiple approaches for satisfactory therapy

11. Dennis Klinman Margaret Schwartz Napoleone Ferrara Mei Zheng Bum-Seok Kim Kaustuv Banerjee Partha Biswas Sujin Lee Colleagues Involved