Open access

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Breast cancer is the leading cause of death by cancer among women and there is an urgent need to improve its
diagnosis and prognosis. MicroRNAs (miRNAs) are non-coding RNAs that regulate gene expression and many
have been implicated in breast cancer. In this article, we focus on circulating miRNAs as biomarkers for breast
cancers and we describe the deregulation of their expression during neoadjuvant chemotherapy (NAC).
Conclusions !!!!!!!!!!!!
1) Accurate biomarkers for minimally invasive diagnosis of primary and metastatic
breast cancer was identified ;
2) NAC induces expression of miR-34a in plasma and tumor tissue, which could be
involved in anti-tumor effect and cardiotoxicity of chemotherapy agents. !
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The expression of 188 plasma miRNAs was determined in 101
patients with primary or metastatic breast cancer and 20 healthy
women. We found 116 miRNAs modified in both primary and
metastatic breast cancer patients compared to controls.
A diagnostic model, based on 8 miRNAs (Fig. 1) measured in 71
breast cancer patients and 14 healthy women, was designed. The
validity of this classification model has then been confirmed on an
independent cohort of 30 patients and 6 controls!
Receiver operating characteristic (ROC) curve derived from the 8-
miRNAs based diagnostic tool exhibited an area under the curves
(AUC) of 0,95 in the validating cohort
Circulating miRNAs expression was assessed in plasma and
tumor of 25 patients before and after the NAC. 25 plasma miRNAs
are significantly modified after NAC (Fig. 2). Tumor suppressor
miR-34a is highly upregulated in plasma at the end of NAC (Fig.
3A). This miRNA displays also an increased expression in the
tumor tissue after NAC in 7 patients with pathological partial
response to NAC (Fig. 3B). This work demonstrates for the first
time that NAC induces expression of miR-34a in plasma and
tumor tissue that might be involved in the anti-tumor effect of the
chemotherapy. Studying the kinetic of miR-34a expression during
NAC revealed that its level is especially increased after
anthracycline-based chemotherapy (Fig. 3C). Moreover, miR-34a
upregulation after anthracycline is correlated with troponin
upregulation in NAC treated patients, suggesting a role of this
miRNA in anthracycline-mediated cardiotoxicity (Fig. 3D).
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Pierre Frères, Claire Josse, Stéphane Wenric, Nicolas Bovy, Meriem Boukerroucha, Ingrid
Struman, Vincent Bours, Guy Jerusalem
GIGA-Research,!!Human!Gene6cs,!!Université!de!Liège!
Contact!:[email protected]!
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20 25 30 35 40
-1
0
1
2
Ct
Ct (NA1 - D-8)
miR-34a
miR-451
miR-122
Upregulated miRNAs
miRNAs P-value FDR Fold-change
miR-106a <0.05 0.08 1.4
miR-122 <0.01 <0.01 1.9
miR-126 <0.01 <0.05 1.3
miR-139-5p <0.001 <0.05 1.5
miR-15b <0.01 <0.05 1.3
miR-197 <0.05 0.1 1.2
miR-199a-3p <0.01 0.1 1.4
miR-223 <0.01 0.09 1.2
miR-328 <0.05 <0.05 1.3
miR-34a <0.0001 <0.001 2.5
miR-374b <0.01 0.07 1.3
miR-423-3p <0.05 <0.05 1.2
miR-425 <0.01 <0.01 1.8
miR-503 <0.01 0.06 1.6
!
Downregulated miRNAs
miRNAs P-value FDR Fold-change
miR-127-3p <0.01 0.08 0.7
miR-150 <0.05 0.06 0.5
miR-29c <0.05 0.1 0.8
miR-324-3p <0.05 0.06 0.6
miR-329 <0.05 0.06 0.6
miR-376a <0.05 0.2 0.7
miR-376c <0.01 0.2 0.7
miR-382 <0.01 0.3 0.7
miR-409-3p <0.01 0.2 0.7
miR-411 <0.05 0.1 0.8
miR-451 <0.01 <0.05 0.5
!
NA1$
D&8$
A B
C
NA1 NA2
D-8
3M
0
10
20
30
40
50
NAC
miR-34a expression (fold-change)
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***
miR-34a expression in plasma
NA1
D-8
0
20
40
60
80
100
NAC
miR-34a expression (2-ΔCt)
miR-34a expression in tumor tissues
*
A B
C
NA1
D-8
0
20
40
60
80
miR-34a expression in plasma
NAC
miR-34a expression (2-ΔCt)
****
D
0.1 110 100 1000
0
1
2
3
miR-34a expression (log10*2-ΔCt)
Troponin T
Correlation between upregulation of
plasma miR-34a and troponins T
miR-122 miR-29c miR-33a
miR-199a-3p
miR-324-3p
let-7d
miR-181c
miR-221
0.0
0.5
1.0
1.5
miRNAs expression (2-ΔCt)
Healthy women
Breast cancer patients
** **** *** ***
** **
**
0.000
0.005
0.010
0.015
0.020
miR-181c expression (2-ΔCt )
****
miRNAs expression in plasma
A
B
AUC = 0.95
sensibility = 0.9
specificity = 0.83
threshold = 0.3
Figure 1 : 8 circulating miRNAs significantly dysregulated in
breast cancer patients
Figure 2 : Circulating miRNAs significantly dysregulated after NAC
Figure 3 : miR-34a upregulation under NAC
NA1 NA2
D-8
3M
0
10
20
30
40
50
NAC
miR-34a expression (fold-change)
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***
miR-34a expression in plasma
NA1
D-8
0
20
40
60
80
100
NAC
miR-34a expression (2-ΔCt)
miR-34a expression in tumor tissues
*
A B
C
NA1
D-8
0
20
40
60
80
miR-34a expression in plasma
NAC
miR-34a expression (2-ΔCt)
****
D
0.1 110 100 1000
0
1
2
3
miR-34a expression (log10*2-ΔCt)
Troponin T
Correlation between upregulation of
plasma miR-34a and troponins T