Influence of Disease Duration on Circulating Levels of miRNAs in Children and Adolescents with New Onset Type 1 Diabetes
Abstract
:1. Introduction
2. Results
2.1. Technical Controls Prior to miRNA Profiling
2.2. miRNA Quantification in 182 Plasma Samples
2.3. Effect of Disease Duration on miRNA Expression Levels during the First Five Years after Diagnosis
2.4. Identification of miRNA Target Genes
2.5. Partial remission and Immunological Status
2.6. Association of the Eight Candidate miRNAs to Cytokines
3. Discussion
4. Materials and Methods
4.1. Study Population and Samples
4.2. Partial Remission (PR) Phase
4.3. Pancreatic Anti-Islet Autoantibodies
4.4. Cytokine Measurements
4.5. miRNA Expression Profiling and Normalization Method
4.6. Validation of Target Genes and Pathway Analysis
4.7. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Study Population | 1 Month | 3 Months | 6 Months | 12 Months | 60 Months |
---|---|---|---|---|---|
n (girls/n) | 40 (24) | 37 (22) | 33 (19) | 34 (20) | 38 (23) |
Age at diagnosis (years) | 8.7 (3.4) | - | - | - | - |
HbA1c (DCCT, %) | 9.1 (1.3) | 7.3 (1.1) | 7.8 (1.6) | 7.9 (1.1) | 8.4 (1.0) |
HbA1c (IFCC,mmol/mol) | 76 (14.2) | 56 (12.0) | 62 (17.5) | 63 (12.0) | 68 (11.0) |
C-peptide (pmol/L) | 629 (371) | 528 (323) | 436 (333) | 257 (251) | 62 (76) |
IDAA1c | 10.8 (2.0) | 9.2 (1.5) | 10.1 (2.2) | 11.0 (1.8) | 12.0 (1.9) |
Insulin dose (units/kg/24 h) | 0.45 (0.27) | 0.47 (0.23) | 0.58 (0.26) | 0.78 (0.27) | 0.88 (0.32) |
Autoantibody positivity (%) | |||||
GAD65A | 55 | 62 | 61 | 56 | 55 |
IA | 80 | 100 | 100 | 100 | 78 |
IA-2A | 73 | 68 | 70 | 68 | 74 |
ICA | 93 | 92 | 94 | 94 | 92 |
ZnT8tripleAb | 65 | 70 | 75 | 76 | 26 |
miRNA | Unadjusted p-Value | Adjusted p-Value |
---|---|---|
hsa-miR-99a-5p | <0.00001 | 0.00022 |
hsa-miR-30e-5p | <0.00001 | 0.00027 |
hsa-miR-497-5p | <0.00001 | 0.00041 |
hsa-miR-10b-5p | 0.00001 | 0.00072 |
hsa-miR-423-3p | 0.00001 | 0.00131 |
hsa-miR-125b-5p | 0.00003 | 0.00252 |
hsa-miR-17-5p | 0.00011 | 0.01057 |
hsa-miR-93-5p | 0.00012 | 0.01094 |
hsa-miR-146a-5p | 0.00094 | 0.08678 |
hsa-miR-484 | 0.00195 | 0.17716 |
hsa-miR-185-5p | 0.00208 | 0.18726 |
hsa-miR-24-3 | 0.00291 | 0.25943 |
hsa-miR-660-5p | 0.00944 | 0.83059 |
hsa-miR-25-3p | 0.01088 | 0.94673 |
hsa-let-7b-5p | 0.01133 | 0.97420 |
hsa-miR-320a | 0.01236 | 0.98546 |
hsa-miR-223-3p | 0.01248 | 0.98546 |
hsa-let-7g-5p | 0.01553 | 0.98546 |
hsa-miR-20a-5p | 0.01815 | 0.98546 |
hsa-let-7i-5p | 0.02298 | 0.98546 |
hsa-miR-142-3p | 0.02412 | 0.98546 |
hsa-miR-32-5p | 0.02544 | 0.98546 |
hsa-miR-486-5p | 0.02989 | 0.98546 |
hsa-miR-142-5p | 0.04400 | 0.98546 |
hsa-miR-320b | 0.04421 | 0.98546 |
hsa-miR-145-5p | 0.04545 | 0.98546 |
hsa-miR-221-3p | 0.04598 | 0.98546 |
hsa-miR-106a-5p | 0.04701 | 0.98546 |
Panther Pathways | Fold Enrichment | p-Value | miRNAs |
---|---|---|---|
Angiogenesis | 3.37 | 8.25 × 10−6 | hsa-miR-10b-5p, hsa-miR-125b-5p, hsa-miR-17-5p, hsa-miR-423-3p, hsa-miR-93-5p, hsa-miR-99a-5p |
Gonadotropin-releasing hormone receptor pathway | 2.9 | 1.77 × 10−5 | hsa-miR-17-5p, hsa-miR-125b-5p, hsa-miR-497-5p, hsa-miR-423-3p, hsa-miR-10b-5p, hsa-miR-93-5p, hsa-miR-30e-5p, hsa-miR-99a-5p |
TGF-β signaling pathway | 4.08 | 7.21 × 10−5 | hsa-miR-10b-5p, hsa-miR-125b-5p, hsa-miR-17-5p, hsa-miR-423-3p, hsa-miR-93-5p, hsa-miR-99a-5p |
CCKR signaling map | 3.11 | 1.07 × 10−4 | hsa-miR-17-5p, hsa-miR-125b-5p, hsa-miR-497-5p, hsa-miR-93-5p, hsa-miR-99a-5p, hsa-miR-10b-5p, hsa-miR-423-3p |
EGF receptor signaling pathway | 3.12 | 1.16 × 10−3 | hsa-miR-10b-5p, hsa-miR-125b-5p, hsa-miR-17-5p, hsa-miR-423-3p, hsa-miR-497-5p, hsa-miR-93-5p, hsa-miR-99a-5p |
PDGF signaling pathway | 2.91 | 3.16 × 10−3 | hsa-miR-10b-5p, hsa-miR-125b-5p, hsa-miR-17-5p, hsa-miR-423-3p, hsa-miR-497-5p, hsa-miR-93-5p, hsa-miR-99a-5p |
Apoptosis signaling pathway | 3.12 | 5.05 × 10−3 | hsa-miR-10b-5p, hsa-miR-125b-5p, hsa-miR-17-5p, hsa-miR-30e-5p, hsa-miR-423-3p, hsa-miR-93-5p |
FGF signaling pathway | 3.05 | 6.90 × 10−3 | hsa-miR-10b-5p, hsa-miR-125b-5p, hsa-miR-17-5p, hsa-miR-423-3p, hsa-miR-497-5p, hsa-miR-93-5p, hsa-miR-99a-5p |
p53 pathway feedback loops 2 | 4.49 | 7.85 × 10−3 | hsa-miR-10b-5p, hsa-miR-125b-5p, hsa-miR-17-5p, hsa-miR-423-3p, hsa-miR-93-5p, hsa-miR-99a-5p |
Ras Pathway | 3.66 | 1.34 × 10−2 | hsa-miR-10b-5p, hsa-miR-125b-5p, hsa-miR-17-5p, hsa-miR-423-3p, hsa-miR-497-5p, hsa-miR-93-5p |
Integrin signalling pathway | 2.41 | 3.07 × 10−2 | hsa-miR-10b-5p, hsa-miR-125b-5p, hsa-miR-17-5p, hsa-miR-423-3p, hsa-miR-497-5p, hsa-miR-93-5p, hsa-miR-99a-5p |
Months after Diagnosis | miRNA | Autoantibody | Spearman Correlation (rs) | Unadjusted p-Value | Adjusted p-Values |
---|---|---|---|---|---|
3 | hsa-miR-99a-5p | ICA | −0.35 | 0.036 | N/S |
3 | hsa-miR-125b-5p | ICA | −0.36 | 0.030 | N/S |
3 | hsa-miR-125b-5p | IA-2A | −0.35 | 0.032 | N/S |
6 | hsa-miR-17-5p | GADA | 0.37 | 0.033 | N/S |
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Samandari, N.; Mirza, A.H.; Kaur, S.; Hougaard, P.; Nielsen, L.B.; Fredheim, S.; Mortensen, H.B.; Pociot, F. Influence of Disease Duration on Circulating Levels of miRNAs in Children and Adolescents with New Onset Type 1 Diabetes. Non-Coding RNA 2018, 4, 35. https://doi.org/10.3390/ncrna4040035
Samandari N, Mirza AH, Kaur S, Hougaard P, Nielsen LB, Fredheim S, Mortensen HB, Pociot F. Influence of Disease Duration on Circulating Levels of miRNAs in Children and Adolescents with New Onset Type 1 Diabetes. Non-Coding RNA. 2018; 4(4):35. https://doi.org/10.3390/ncrna4040035
Chicago/Turabian StyleSamandari, Nasim, Aashiq H. Mirza, Simranjeet Kaur, Philip Hougaard, Lotte B. Nielsen, Siri Fredheim, Henrik B. Mortensen, and Flemming Pociot. 2018. "Influence of Disease Duration on Circulating Levels of miRNAs in Children and Adolescents with New Onset Type 1 Diabetes" Non-Coding RNA 4, no. 4: 35. https://doi.org/10.3390/ncrna4040035