1. Cancer and Apoptosis combined An historical introduction Mathieu Noteborn LIC, Leiden University, NL LC Workshop - 29 October 2007

2. Cancer and Apoptosis combined An historical Introduction Apoptosis versus cancer Viruses inducing Cancer: lessons to be learned Proteins killing tumor cells Apoptin E4orf4 HAMLET MDA7 NS1 TRAIL A brief preview to our workshop

3. Apoptosis and life belong together

4. Apoptosis is a genetically regulated network of pathways Factors can “positively or negatively” interfere with apoptosis machinery Imbalances in apoptosis pathways underlie many different diseases

5. West Nile virus induces apoptosis in brain cells resulting in meningitis/encephalitis Inhibition of apoptosis results in tumor cell formation Interference in apoptosis machinery Induction Inhibition

6. Interference at various steps in the apoptotic machinery results in tumor formation

7. “Living with p53, dying of p53” Aylon & Oren, 2007 Mutated p53/partners might result in genetic instability/cancer Cellular background Cellular background

8. Up-regulation of anti-apoptotic proteins results in tumor formation

9. Bcl-2/BclXl form dimers via their BH3 domains with each other or with pro-apoptotic family members

10. EpiGalleCatechin Gallate; EGCG Reed, 2006 Prevention of formation of anti-apoptotic Bcl-2 homo- or hetero-dimers results in killing of tumor cells anti-apoptotic proteins as Bcl-2 are potential drug targets

11. Expression of Livin causes chemo-resistance of tumor cells

12. Liu et al., 2007 Down-regulation of Livin sensitizes tumor cells to chemotherapeutic drugs

13. Aberrancies in Bcl-2, p53 & Fas: tumor “beats” our defense mechanism

14. Apoptosis versus Cancer Summary Defects in apoptosis pathways Contribute to cancer Permitting aberrant cell behavior Desensitizing tumor cells to: Immune-mediated attack Chemotherapy Cancer development and anti-cancer therapy

15. Cancer and Apoptosis combined An historical Introduction Apoptosis versus cancer Viruses inducing Cancer: lessons to be learned Proteins killing tumor cells Apoptin E4orf4 HAMLET MDA7 NS1 TRAIL A brief preview to our workshop

16. DNA tumor viruses Papilloma virus Adenovirus Herpesvirus Epstein Barr virus Hepatitis B virus Polyoma virus SV40 RNA tumor viruses Retroviruses

17. SV40 transforms cells SV40 LT interacts with tumor suppressor p53

18. DNA-tumor viruses express proteins interfering with cellular factors resulting in cellular transformation O’Shea, 2004

19. SV40 LT and st have transforming activities LT interacts with e.g. p53, Rb st interacts with PP2A Pipas

20. PP2A-dependent pathways targeted by SV40 st contributes to cell transformation Sontag & Sontag, 2006 Arroyo

21. …. and most likely much more gene products are affected by st synthesis Affymetrix studies SV40 st affects e.g. expression of factors within the Wnt, Hedgehog & Notch pathways Ali-Seyed et al., 2006

22. SV40 st affects Apoptosis Pathways Down-regulation expression of pro-apoptotic factors Up-regulation expression of anti-apoptotic factors

23. DNA tumor viruses & apoptosis SV40 transforming LT and st Inhibition of pro-apoptotic processes (p53/caspase) Stimulation of anti-apoptotic processes (survivin) Adenovirus proteins Inhibition of pro-apoptotic processes Stimulation of anti-apoptotic processes ?

24. White, 2006 Transforming adenovirus inhibits and induces apoptosis

25. SV40 LT/st inhibits apoptosis Adenovirus E1B proteins inhibit apoptosis Adenovirus E1A induces apoptosis Why does transforming adenovirus harbor E1A protein? Apoptosis is an essential step in (Ad) transformation? Next Oncolytic viruses require transformation events to induce cell death/apoptosis

26. Oncolytic Viruses E.g. Parvovirus H1, Newcastle Disease Virus, (engineered) Adenovirus Vähä-Koskela et al., 2007

27. Molecular basis for selective permissiveness of cancer cells to oncolytic viruses - Chromosomal instability - Apoptosis resistance - Higher levels of virus receptors (e.g. ICAM-1) - Impaired anti-viral response ( lack of critical IFN response ) - Adenovirus lacking E1B grows only in p53-minus cells

28. Using viruses in conjunction with other forms of treatment Nuesch/Rommelaere

29. Viruses inducing cancer: lessons to be learned Summary Viruses transform cells by inhibiting apoptosis SV40 Viruses transform cells by inhibiting & inducing apoptosis Adenovirus + others? Oncolytic viruses “use” transforming events to kill cells Parvovirus H1 (NS1 protein) Newcastle disease virus

30. Cancer and Apoptosis combined An historical Introduction Apoptosis versus cancer Viruses inducing Cancer: lessons to be learned Proteins killing tumor cells Apoptin E4orf4 HAMLET MDA7 NS1 TRAIL A brief preview to our workshop

31. Transforming events are needed for proteins inducing tumor-specific apoptosis: Use of a paradox? DNA damage ApoptosisArrest Apoptosis Survival Pathway interference affecting cell transformation Apoptin, E4orf4, HAMLET, NS1, MDA7, TRAIL All seem to sense tumor-specific processes Abrahams, unpublished

32. Transformation events activate apoptin Apoptosis Normal cells + LT/st N localization Apoptin + LT Apoptin a-108P Phosphorylation Peng SV40 LT/st are transforming proteins

33. E4orf4 affects directly tumor-related PP2A PP2A Lavoie

34. Tumor factors encountered by HAMLET - MDA7 – TRAIL TRAIL MDA7 HAMLET Svanborg Ramesh Walczak & Borst

35. All Participants are invited to send in their contribution to the book “Proteins Killing Tumour Cells” Within the Research SignPost Series on Cell Death and Disease Editors Backendorf – Borst - Noteborn – Tavassoli

36. Organizers C Backendorf, J Borst M Noteborn & M Tavassoli Advisory Board members J Brouwer, M Green, F Farzaneh & AJ van der Eb WISH YOU A VERY FRUITFUL LC-WORKSHOP

37. References Ali-Seyed et al., 2006, BMC Cancer 6, 54 Aylon & Oren, 2007, Cell 130, 597 Kooistra et al., Nov. 2007, MRMC, in press Liu et al., 2007, Cancer Letters 250, 168 Reed, 2006, Nature Clinical Practice 3, 388 Sontag & Sontag, 2006, Cell. Mol. Life Sciences 63, 2979 Vähä-Koskela et al., 2007, Cancer Letters, 254, 178 White, 2006, Cell Death & Differentiation 13, 1371