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
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
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)
Progression-Free Survival
Prob
abili
ty o
f Pr
ogre
ssio
n-Fr
ee S
urvi
val
Time From Baseline (Months)0 2 4 6 8 10 12 14 16 18 22 24 26 28 30
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
20
Log rankP = .0001
7.0 months2.7
months
≥ 5 CTCs/7.5 mLn = 87 (49%)
< 5 CTCs/7.5 mLn = 90 (51%)
Cristofanilli M, et al. N Engl J Med. 2004;351:781-791.
Prognostic Value of Baseline CTC Counts
Overall Survival
Time From Baseline (Months)0 2 4 6 8 10 12 14 16 18 22 24 26 28 30
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
20
Log rank P < .000121.9 months
10.9months
≥ 5 CTCs/7.5 mLn = 87 (49%)
< 5 CTCs/7.5 mLn = 90 (51%)
Cristofanilli M, et al. N Engl J Med. 2004;351:781-791.
Prognostic Value of Baseline CTC Counts
Prob
abili
ty o
f Pr
ogre
ssio
n-Fr
ee S
urvi
val
64%
43%
< 5 CTCsn = 90 (51%)
Cristofanilli M, et al. ASCO 2005. Abstract 524.
Probability of Death and Baseline CTC Counts
Time From Baseline (Months)0 2 4 6 8 10 12 14 16 18 22 24 26 28 30
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
20
29%
≥ 5 CTCsn = 87 (49%)
53%
12%19%
Prob
abili
ty o
f Dea
th
Change in CTC Count During Therapy Predicts Overall Survival
Cristofanilli M, et al. ASCO 2005. Abstract 524.
4
1 vs 4 P < .0001
4 > 5 CTCs at all time points 39 (22%)
2
1 vs 2 P = .3188
2 vs 4 P < .0001
2 > 5 at baseline and < 5 CTC at last draw 38 (21%)
3
1 vs 3 P = .0014
3 vs 4 P = .0051
2 vs 3 P = .0397
3 < 5 at Early Draw and > 5 CTC at last draw 17 (10%)
Time From Baseline (Months)0 2 4 6 8 10 12 14 16 18 22 24 26 28 30
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
20
1
1 <5 CTCs at all time points 83 (47%)
Prob
abili
ty o
f Sur
viva
l
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
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
1. 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
Human equilibrative nucleoside transporter 1Deoxycytidine kinase (dCK)
CK8
Spiked MCF7 cells in 10 ml of blood
PRIMER SEQUENCE 5’→3’ Annealing(t°C)
SIZEbp
GAPDH upstream ACATGTTCCAATATGATTCC 60 161
downstream TGGACTCCACGACGTACTCAG
CD 45 upstream CAGCGCTTCCAGAAGGGCTC 60
downstream CCGTGCAGCTCTACGAGAGG
CK-8
upstream ACTGAGATCTCAGAGATCAA
60downstream AATACTCATGTTCTGCATCC
MRP 1 upstream CTGACAAGCTAGACCATGAATGT 60 353
downstream TCACACCAAGCCGGCGTCTTT
MRP 2 upstream GTGATGGTGCTTGTAATCCCA 60 277
downstream TGACCACAGATACCAGGACTG
MRP 4 upstream GGATCCAAGAACTGATGAGTTAAT 65 358
downstream TCACAGTGCTGTCTCGAAAATAG
MRP 5
upstream CTAGAGAGACTGTGGCAAGAAGAGC 68 332
downstream AAATGCCATGGTTAGGATGGC
MRP 7 upstream TAGGCACTGACTCTGAACGG 60 160
Downstream TTGTTGACGGGTACCAGCAG
dCK upstream TGCAGGGAAGTCAACATT 52 146
downstream TCCCACCATTTTTCTGAG
hENT 1 upstream GCTTGAAGGACCCGGGGAGC 67 502
downstream TGGAGAAGGCAAAGGCAGCCA
Sensibilità:bassa espressione MRP
Elevata espressione hENT/dCK
Resistenza:elevata espressione MRP
Bassa espressione hENT/dCK