1. Immune Response and Fighting Brain Cancer with Gene Therapy Abbie Hunt November 10, 2009

2. Overview Glioblastoma multiforme (GBM) Dendritic Cells (DC) Fms-like tyrosine kinase 3 ligand (Flt3L) Toll-like receptors (TLR) Bone-marrow derived DCs (BMDC) High-mobility-group box 1 (HMGB1)

3. Cell lines, etc. Challengers Ad-TK Ad-Flt3L Ad0 Returning Champions GL26 GL261 B16-F10 LLc1

4. Mice! Female C57BL/6 MyD88 -/- TLR4 -/- WT Chimeric WT and GFP +/+ WT and TLR2 -/- http://brainwindows.files.wordpress.com/2008/10/image-gfp-mouse-crop-copy.jpg

5. Glioma Models GL26 GL261 B16-F10 Ad0 Ad-Flt3L + Ad0 Ad-TK + Ad0 Ad-Flt3L + Ad-TK GCV Saline Rechallenged

6. T cells necessary in tumor regression

7. DCs come from bone marrow

8. TLR2 expression by BMDC necessary in tumor regression

9. HMGB1 mediates TLR2-dependent glioma tumor regression Increased levels of HMGB1 from GL26 treated with Ad-TK+GCV in vitro Increased serum levels in mice Blocked with glycyrrhizin or specific anti- HMGB1 antibodies

10. HMGB1 released from tumor cells with Ad- TK+GCV, chemotherapy, or radiation therapy

11. Ad-TK(+GCV) + Ad-Flt3L = Survival and Increased HMGB1

12. Recap T cells necessary in tumor regression DCs come from bone marrow TLR2 expression by BMDC necessary in t.r. HMGB1 mediates TLR2-dependent glioma t.r. HMGB1 released from tumor cells with Ad- TK+GCV, chemotherapy, or radiation therapy Ad-TK (+GCV) and Ad-Flt3L therapy results in GL261 and B16-F10 tumor regression and increased HMGB1 levels

14. References 1. Curtin J.F., Liu N., Candolfi M., et al. 2009. HMB1 Mediate Endogenous TLR2 Activation and Brain Tumor Regression. PLoS Medicine. 6(1): 84-104. 2. Southgate T., Kroeger K.M., Liu C., Lowenstein P.R. (2008). Gene transfer into meural cells in vivo using adenoviral vectors. Published online at Wiley Interscience. Available online at http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2659706/ Accessed 11/4/09. 3. Sandmair A., Vapalahti M., Yla-Herttuala S. (2000). Adenovirus- Mediated Herpes Simplex Thymidine Kinase Gene Therapy for Brain Tumors. Cancer Gene Therapy: Past Achievements and Future Challenges. Plenum Publishers, New York. 163-170. 4. Stevenson, P. Immunological Memory. The Australian Naturopathic Network. Revised 5/18/2002. Available online at http://www.ann.com.au/MedSci/immunolo.htm Accessed 11/3/09. http://www.ann.com.au/MedSci/immunolo.htm 5. Multitudes of Wikipedia pages