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Table 1. Sample Characteristics (n = 68)
Age
Sex
Stage
Outcome
MYCN
Sample
Plasma
Tissue samples
(corresponding samples)1
Total
31
37 (27)
<=12 m
10
12 (9)
13 -17 m
5 (3)
≥ 18m
17
20 (15)
Male
22
25 (19)
Female
12 (8)
L1
11
12 (10)
L2
8 (5)
11
15 (11)
MS
2 (1)
Dead
10
16 (10)
Alive
21
21 (17)
Non-amplified
25
26 (21)
Amplified
11 (6)
( ) = available corresponding tissue samples to plasma samples
Figure 1. Workflow of the current study. Stage group : INRGSS stage M and non M. Outcome group : deceased and alive.
MYCN group : amplified and non-amplified.
A.
Normalized expression
2500
p = 0.017
2000
1500
1000
500
Alive outcome Deceased
B.
1800
1600
Tissue
1400
1200
1000
800
Alive
600
Deceased
400
200
500
1000
1500
2000
2500
Plasma
Figure 2. Results of analysis in outcome group.
A. Plasma level of the miRNA significantly correlated with outcome,
miR-92a-3p.
B. Correlation plot of miR-92a-3p levels in plasma and tissue samples.
Spearman rho = 0.308.
A.
2000
P = 0.002
Normalized expression
1800
1600
1400
1200
1000
800
600
400
200
L1, L2, MS
stage
B.
900
800
Tissue
700
600
500
400
L1, L2 & MS
300
200
100
500
1000
1500
2000
Plasma
Figure 3. Results of analysis in INRGSS stage group. A. Plasma level
of the miRNA correlated with INRGSS stage, miR-375.
B. Correlation plot of miR-375 levels in plasma and tissue samples.
Spearman rho = 0.545.
10
15
20
25
500
1000
1500
2000
2500
MYCN
Ampilified
Non-Amplified
Amplified
Non-AmplifiedMYCN
p = 0.006
p = 0.007
D.
B.
18
16
14
12
10
1800
1600
1400
1200
1000
800
600
400
200
500
Plasma
1500
Plasma
1000
Non-amplified
2000
20
Non-amplified
Amplified
2500
Amplified
Figure 4. Plasma level of the miRNA significantly correlated with MYCN status, miR-92a-3p. B. Correlation plot of miR-92a-3p levels in plasma
and tissue samples. Spearman rho = 0.255. C. Plasma level of the miRNA significantly correlated with MYCN status, miR-99a-5p.
D. Correlation plot of miR-99a5p levels in plasma and tissue samples. Spearman rho = 0.212.
C.
Normalized expression
Normalized expression
Tissue
Tissue
A.
Figure 5. Receiver operating characteristic curve of the combination of miR-92a-3p, miR-375, and miR-99a-5p levels in
plasma samples (AUC = 0.726, p = 0.001, 95 % CI = 0.612-0.841, sensitivity = 77 %, specificity = 56.7 %)
Figure 6. Kaplan-Meier plot of outcome based on miR-92a-3p levels. The 5-year overall survival rate was 89% for
patients with low miR-92a-3p expression levels and 45 % for patients with high miR-92a-3p expression levels.
Low/high expression levels were defined as expression levels below or above the average of plasma miR-92a-3p levels
in all cases (mean = 732.15 counts).
...