CirculatingTumor Cells: Toward a clinical benefit?

1. CirculatingTumor Cells:Toward a clinical benefit? Giuseppe Naso MD, PhD, Associated Professor of Medical Oncology Director of Traslational Oncology Paola Gazzaniga, MD, PhD , University Researcher in Traslational Oncology Sapienza Università di Roma

2. Metastatic Breast Cancer MBC is a heterogeneous condition Treatment is palliative Current clinical, laboratory, and radiological methods inadequate for Assessment of prognosis Definition of biology Measurement of efficacy/treatment benefit

3. Prognostic and Predictive Value of CTCs in MBC Hypothesis • Measurement of CTCs in MBC may: • Identify aggressive disease (prognostic value) • Provide early determination of treatment efficacy/benefit (predictive value)

5. ≥ 5 CTCs/7.5 mL Prognostic Value of Baseline CTC Counts Progression-Free Survival 100% 90% 80% < 5 CTCs/7.5 mL 70% n = 90 (51%) 60% Probability of Progression-Free Survival 7.0 months 50% 2.7months 40% 30% Log rank 20% P = .0001 10% n = 87 (49%) 0% 0 8 14 16 18 20 22 24 26 28 2 6 10 12 4 30 Time From Baseline (Months) Cristofanilli M, et al. N Engl J Med. 2004;351:781-791.

6. < 5 CTCs/7.5 mL n = 90 (51%) ≥ 5 CTCs/7.5 mL Prognostic Value of Baseline CTC Counts Overall Survival 100% 90% 80% 70% 60% Log rank P < .0001 21.9 months Probability of Progression-Free Survival 50% 10.9months 40% 30% 20% n = 87 (49%) 10% 0% 0 8 14 16 18 20 22 24 26 28 2 4 6 10 12 30 Time From Baseline (Months) Cristofanilli M, et al. N Engl J Med. 2004;351:781-791.

7. Probability of Death and Baseline CTC Counts 100% ≥ 5 CTCs 90% n = 87 (49%) 80% 70% 64% 60% 53% Probability of Death 50% 43% 40% < 5 CTCs 29% 30% n = 90 (51%) 19% 20% 12% 10% 0% 0 8 14 16 18 20 22 24 26 28 2 6 10 12 4 30 Time From Baseline (Months) Cristofanilli M, et al. ASCO 2005. Abstract 524.

8. Change in CTC Count During Therapy Predicts Overall Survival 1 <5 CTCs at all time points 83 (47%) 2 > 5 at baseline and < 5 CTC at last draw 38 (21%) 3 < 5 at Early Draw and > 5 CTC at last draw 17 (10%) 100% 4 > 5 CTCs at all time points 39 (22%) 90% 1 vs 2 P = .3188 80% 1 vs 3 P = .0014 70% 1 vs 4 P < .0001 60% Probability of Survival 50% 2 1 40% 2 vs 3 P = .0397 30% 20% P < .0001 2 vs 4 10% 3 vs 4 P = .0051 4 3 0% 0 8 14 16 18 20 22 24 26 28 2 6 10 12 4 30 Time From Baseline (Months) Cristofanilli M, et al. ASCO 2005. Abstract 524.

9. CTC Significance in MBC: Prognostic and Predictive Values • Prognosis — strongest independent factor • In newly diagnosed disease (first-line treatment) • Over the course of the disease (second-, third-, or higher-line treatment) • Prediction — predict treatment efficacy at 3-4 weeks • May represent a new standard of care

10. 3 key points in metastatic breast cancer 1.Una elevata conta delle CTC (>5/7.5 ml) è associata a ridotta sopravvivenza globale (10 vs 22 mesi) e ridotto intervallo libero da malattia (2 vs 8 mesi) 2. La conta delle CTC è fattore prognostico più affidabile delle indagini radiologiche e dello stato recettoriale nei k mammari 3. 1/3 delle pazienti con k mammario HER-2 negativo hanno CTC HER-2 positive

11. 2 key points in early breast cancer • La risposta alla terapia adiuvante è associata ad una riduzione • del numero di CTC 2. La presenza di CTC 4 settimane dopo la chirurgia correla con un ridotto intervallo libero da malattia e con ridotta sopravvivenza globale

16. possiamo caratterizzare il loro profilo di chemiosensibilità?????

18. Human equilibrative nucleoside transporter 1 Deoxycytidine kinase (dCK)

20. qual'è il nostro scopo? profilo di chemiosensibilità sulle CTC verificare se tale profilo correli con la risposta clinica alla chemioterapia

23. cellule epiteliali???? 5-10 mononuclear cells/ml

24. CTC= CD45- EpCam+ CK8+

25. assay sensitivity test CK8 Spiked MCF7 cells in 10 ml of blood

26. moriranno in circolo????

27. CTC= CD45- EpCam+ CK8+ survivin +

29. Sensibilità: bassa espressione MRP Elevata espressione hENT/dCK Resistenza: elevata espressione MRP Bassa espressione hENT/dCK