Egal Gorse 2006-2007

1. Transfection of Recombinant Plasmid DNA From Immunotoxins into Eukaryotic Cells for the Development of an Antibody Targeted Cancer Therapy Egal Gorse 2006-2007

2. Goals • To determine if mammalian carrier cells would accept and express immunotoxin DNA. • To evaluate which carrier cells most effectively expressed immunotoxin DNA.

3. Cancer • Cancer is the leading cause of death in the world (7.6 million per year). • There is no effective treatment for cancer. • Chemotherapy is inefficient (resistance and not targeted). • Immunotoxins have shown promise as an effective cancer therapy (Vallera 2005).

7. How EpCAMKDEL Kills Cells A T B Binding Outside Inside Cytosol Golgi Apparatus Endoplasmic Reticulum

8. How EpCAMKDEL Kills Cells A T B Outside Inside Transport Vesicle Cytosol Golgi Apparatus Endoplasmic Reticulum

9. How EpCAMKDEL Kills Cells A T B Outside Inside Transport Vesicle Cytosol Golgi Apparatus Endoplasmic Reticulum

10. How EpCAMKDEL Kills Cells A T T B Outside Inside Cytosol Transport Vesicle Golgi Apparatus Endoplasmic Reticulum

11. How EpCAMKDEL Kills Cells T Outside Inside Cytosol A T B Golgi Apparatus Transport Vesicle Endoplasmic Reticulum

12. How EpCAMKDEL Kills Cells T Outside Inside Cytosol Golgi Apparatus A T B Endoplasmic Reticulum

13. How EpCAMKDEL Kills Cells B T Outside Inside Cytosol Golgi Apparatus Endoplasmic Reticulum

14. How EpCAMKDEL Kills Cells A T T Outside Inside Cytosol Golgi Apparatus A Endoplasmic Reticulum

15. How EpCAMKDEL Kills Cells T Outside Inside Cytosol Golgi Apparatus Transport Vesicle Endoplasmic Reticulum A

16. How EpCAMKDEL Kills Cells A T Outside Inside Cytosol Golgi Apparatus Endoplasmic Reticulum

17. How EpCAMKDEL Kills Cells A T Outside Inside Cytosol Golgi Apparatus Endoplasmic Reticulum

18. Conventional Immunotoxin Administration

19. Cells Transfected With Immunotoxins Directly at the Site of The Tumor Therapy Targeted Directly at The Site Of The Tumor

20. Transfecting EpCAMKDEL Immunotoxin DNA Into Cancerous Cells Using LNCX.NGFR

21. Transfecting EpCAMKDEL Immunotoxin DNA Into Cancerous Cells Using LNCX.NGFR

22. Transfecting EpCAMKDEL Immunotoxin DNA Into Cancerous Cells Using LNCX.NGFR

23. Transfecting EpCAMKDEL Immunotoxin DNA Into Cancerous Cells Using LNCX.NGFR

24. Transfecting EpCAMKDEL Immunotoxin DNA Into Cancerous Cells Using LNCX.NGFR

25. Transfecting EpCAMKDEL Immunotoxin DNA Into Cancerous Cells Using LNCX.NGFR

26. NIH-3T3 Transfected with EpCAMKDEL/LNCX.NGFR

27. Proliferation Assay

28. Proliferation Assay

29. Conclusions • Immunotoxin genes were capable of transfection into mammalian cells. • The mammalian carrier cells were capable of expressing the immunotoxin genes. • Expressed immunotoxins were capable of killing cancerous cells. • The mouse cell lines tested were more capable of transfection than the human cell lines.

30. Future Study • Testing in a live model (in vivo). • Use results in an effective cancer therapy for human.

31. Acknowledgments • Team Research • Dr. Daniel Vallera • Dr. Deborah Todhunter • Ms. Lois Fruen

32. Transfection of Recombinant Plasmid DNA From Immunotoxins into Eukaryotic Cells for the Development of an Antibody Targeted Carcinoma Therapy Egal Gorse 2006-2007