Notch and Cancer

1. Notch and Cancer IPO-LISBOA CIPM Angiogenesis group Francisco Caiado Sérgio Dias

2. The Hallmarks of Cancer(Hanahan and Weinberg) Tissue Invasion and Metastasis

3. Cancer Hallmarks and Notch signaling Tissue Invasion and Metastasis Modulation of the Notch Signaling Pathway

4. Notch-Delta signaling pathway • Regulates: • establishment of patterns of gene expression; • cell differentiation; • regulates binary cell fate choice; • maintenance of stem cell populations; • Function: • Embryonic Development; • Adult Self-Renewing Organs; CANCER Roca, C. and Adams, R. Genes & Dev. 2007 21: 2511-2524

5. Abnormal Notch signaling and cancer Oncogenic activity of Notch Tumor supressor activity of Notch Maillard,I. and Pear, W. Cancer Cell 2003

6. Abnormal Notch signaling and cancer • Targeting Notch signaling: • γ-secretase inhibitors (GSIs) are in early clinical trials; • mAbs targeting the ‘negative regulatory region’ (NRR) of notch; • mAbs that against DLL4 inhibit Notch signaling in endothelial cells and cause non-functionaltumor angiogenesis; De la Pompa et al. Endocrine Reviews 28(3):339–363

7. Cancer Hallmarks and Notch signaling Tissue Invasion and Metastasis Modulation of the Notch Signaling Pathway

8. Tumor Angiogenesis • Pro-angiogenic factors: • VEGF, FGF, Neuropillin, Ang-2, MMPs… • Vessel stabilizing factors: • Notch-Delta, PDGF-1, Ang-1, ECMs… Hashizume, H.l NCR 2000

9. Notch signaling and tumor angiogenesis Ridgway, J. et al Nature 2006 Thuston, G. et al NCR 2006

10. Notch signaling pathway? Post-natal vasculogenesis • Bone marrow (BM) derived progenitor cells: • recruited during physiological and “malignant” angiogenesis; • BM mobilization; homing to angiogenic sites (Integrins); invasion and migration; • Induction of angiogenesis: • Differentiation into endothelial cells; • Activation of pre-existing endothelial cells; Rafii, S. et al NCR 2002

11. 1. Notch signaling regulates BM-progenitor endothelial differentation? Markers: CD133+CD34+KDR+ Lin- Sca-1+ Flk-1+ Igreja, C. et al Exp. Hematol. 2006 Caiado, F. et al Plos One 2008

12. Notch signaling inhibition impairs adhesion and integrin expression Caiado, F. et al Plos One 2008

13. 2. BM-progenitor modulate endothelial activation via Notch signaling? Subcutaneous injection of human or mouse tumors Transplant of BM-progenitors control or with reduced Dll4 • Tumor growth; • Tumor apoptosis; • Vessel density; • Vessel stability; Sub – lethal irrad. NOD-SCID female

14. ** C 60 * WT-EPC 50 Dll4-EPC 12 40 Apoptosis index 10 30 Tumor Volume (mm3) 8 20 10 6 0 4 Control wtEPC Dll4+/-EPC 2 0 18 0 14 22 26 Days BM progenitors with reduced Dll4 decrease tumor proliferation and increase apoptosis Real, C Submited 2008

15. Pericyte coverage : Hypoxic index : BM progenitors with reduced Dll4 induce increased but non-functional vascularization Vessel Density : Real, C Submited 2009

16. EPC Activated endothelial cell Endothelial cell Apoptotic endothelial cell Pericyte Apoptotic pericyte Notch signaling modulates BM-progenitor function during tumor angiogenesis • Notch signaling regulates BM-progenitor cell endothelial differentiation; • Dll4 expressed on BM-progenitor cells regulates endothelial stabilization during tumor angiogenesis;

17. Acknowledgments: S Dias Francisco A. Gomes A. Cachaco Carla Tânia A. Costa Cheila Cristina Jacinta Cristiana Sara • (2008/2009) Angiogenesis Lab members (Leonor Remédio Missing); • Dr.Antonio Duarte (group members), Dr.Yadgita Hideo (group members); • FCT, GlaxoSmithKline, Fundação Calouste Gulbenkian;