Eotaxin: a Potential Target for Therapy in Chronic Rhinosinusitis

1. Eotaxin: a Potential Target for Therapy in Chronic Rhinosinusitis Alisha N. West, MD Grand Rounds University of California, Los Angeles December 1, 2010

2. History • Chronic Rhinosinusitis (CRS) is characterized by signs and symptoms of inflammation • Stimulated by environmental pathogen • Chronic epithelial stimulation  immune response with a subset of patients exhibiting eosinophilia • Lack of effective mucociliary clearance • Currently, paucity of effective therapies

3. History • Chronic Rhinosinusitis is one of the leading causes of annual visits to the Otolaryngologist • 66 million adults in the US reported sinus problems last year • $5.8 billion dollars is spent annually on treatment for CRS • Symptoms usually recur within three months of therapy

4. Hypothesis Eotaxin plays a role in CRS inflammation and could serve as a potential target for therapy

5. Background • Eotaxin-1 and 2 implicated in inflammation seen in ABPA and asthma • IL-13 and IL-4 up-regulate production of Eotaxin by epithelial cells • Eotaxin is a chemotactic factor that attracts eosinophils to the site of inflammation • Eosinophils release MBP which has been shown to cause epithelial cell damage in ABPA patients • Toxic levels of MBP have been found in the mucus of CRS patients • Prior to our current project, the role of eosinophils and eotaxin had not been examined in CRS

6. Materials and Methods • UNC IRB approved protocols • Sinonasal mucosa harvested during endoscopic sinus surgery60 subjects • Tissue embedded in parrifin and H&E stain performed for EOS • Cell culture protocol • Cellular disaggregation • Plated in serum-free media until confluent, dispersion in trypsin • Re-plated on Milicells™ in air-liquid interface media at density of 250,000 cells/Millicell™ • After confluent monolayeraspirate apical media • Washed and re-fed basally for 21 days until ciliated and mucus production • 4 cell cultures/specimen • Apical challenge with Pseudomonas, Staph, and TSB • Basal Media collected, washed and replaced daily 0-96 hours

7. IL-1beta IL-2 IL-4 IL-5 IL-6 IL-10 IL-12p70 TNF-alpha IFN-gamma G-CSF IP-10 GM-CSF IL-9 IL-13 KC MCP-1 MIP-1a RANTES IL-1a IL-7 IL-15 IL-17 Materials and MethodsLuminex 22-plex battery of Cytokines

8. Materials and Methods • Alveolar Lavage has demonstrated eosinophilia in ABPA and asthma patients as well as animal models • Endosinus lavage with evaluation of eosinophilia in CRS patients and normal volunteers • Complete cell count and Eosinophil count performed with cytospin

9. Tissue Harvest

10. Results

11. H&E Stain 10xCRS Control

12. EosinophiliaCRS Control

13. Eosinophilia in Endosinus Mucosa

14. Eotaxin ELISA

15. Luminex Results

16. Conclusions • There is significant Eosinophilia in CRS endosinus epithelium when compared to normal controls • CRS epithelium produces a higher concentration of eotaxin at baseline and after an infectious challenge when compared to controls • Eotaxin should be investigated as a potential target for therapy in CRS

17. References

18. Thank You