Quantitative Prediction of Steatosis in Patients with Non-Alcoholic Fatty Liver by Means of Hepatic MicroRNAs Present in Serum and Correlating with Hepatic Fat
Abstract
:1. Introduction
2. Results
2.1. Human Liver and Serum MicroRNA Sequencing and Building of Steatosis Predictive Models
2.2. Biological Significance of the 19 miRNAs Identified Both in Liver and Serum That Show Correlation with Liver Steatosis
2.3. PLS Model Based on Only 2-3 Serum miRNAs to Predict % of Fat in NAFLD Patients
3. Discussion
4. Materials and Methods
4.1. Patients and Human Samples
4.2. Quantification of Intrahepatic TG and Total Lipids
4.3. RNA Isolation, Small RNA Library Preparation and miRNAseq Analysis
4.4. Bioinformatics Analysis and Modeling
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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miRNA | Guide Passenger (1/0) | Liver R (p-Value) | Serum R (p-Value) | Lipid Metab. Target Genes | L/S Ratio * | Penalty Score | |
---|---|---|---|---|---|---|---|
mirPath microTcds | mirDIP Target Scan | ||||||
miR-10a-5p | 1 | 0.61 (0.003) | −0.65 (0.0007) | 18 | 20 | 99 | 0 |
miR-98-5p | 1 | −0.49 (0.02) | 0.74 (0.0001) | 29 | 39 | 48 | 0 |
miR-19a-3p | 1 | −0.56 (0.008) | 0.61 (0.001) | 36 | 39 | 63 | 0 |
miR-30e-5p | 1 | −0.61 (0.003) | 0.44 (0.03) | 40 | 31 | 12 | 0 |
miR-32-5p | 1 | −0.45 (0.03) | 0.51 (0.01) | 42 | 36 | 14 | 0 |
miR-145-5p | 1 | 0.62 (0.002) | −0.54 (0.007) | 8 | 23 | 104 | −1 |
let-7d-5p | 1 | 0.64 (0.002) | 0.49 (0.02) | 15 | 43 | 2 | −1 |
miR-181c-5p | 1 | 0.70 (0.0003) | −0.47 (0.02) | 12 | 41 | 6 | −1 |
miR-23a-3p | 1 | 0.70 (0.0004) | −0.47 (0.02) | 36 | 34 | 6 | −1 |
let-7b-5p | 1 | 0.61 (0.003) | 0.43 (0.04) | 29 | 41 | 4 | −1 |
miR-148a-3p | 1 | −0.59 (0.004) | 0.23 (0.3) | 31 | 51 | 331 | −1 |
miR-191-5p | 1 | 0.47 (0.03) | −0.49 (0.01) | 0 | 2 | 3 | −2 |
miR-769-5p | 1 | 0.69 (0.0005) | −0.47 (0.02) | 3 | 0 | 6 | −2 |
mir-140-3p | 1 | 0.68 (0.0006) | −0.41 (0.05) | 19 | 0 | 5 | −2 |
mir-660-5p | 1 | −0.58 (0.005) | −0.40 (0.06) | 8 | 0 | 21 | −2 |
miR-335-5p | 1 | −0.29 (0.2) | 0.28 (0.2) | 9 | 22 | 11 | −3 |
mir-30a-3p | 0 | 0.56 (0.007) | 0.58 (0.003) | 8 | 0 | 189 | −3 |
miR-136-3p | 0 | −0.62 (0.002) | 0.42 (0.04) | 3 | 0 | 62 | −3 |
miR-17-3p | 0 | −0.51 (0.02) | −0.17 (0.4) | 17 | 0 | 11 | −5 |
RMSECV | miRNA 1 | miRNA 2 | miRNA 3 |
---|---|---|---|
4.4 | miR-98-5p (2.55) | miR-19a-3p (2.25) | miR-145-5p (−2.00) |
4.4 | miR-98-5p (2.81) | miR-32-5p (2.05) | miR-145-5p (−2.20) |
4.5 | miR-30e-5p (1.73) | miR-98-5p (3.14) | miR-145-5p (−2.46) |
4.5 | miR-98-5p (3.75) | miR-145-5p (-2.93) | |
4.6 | miR-98-5p (3.75) | miR-32-5p (2.73) | |
4.7 | miR-98-5p (3.13) | miR-19a-3p (2.76) |
Mean ± SD | Min | Max | CV% | |
---|---|---|---|---|
µg Liver TG/mg prot | 919 ± 494 | 210 | 1948 | 60% |
µg Liver lipids/mg prot | 730 ± 539 | 107 | 1894 | 70% |
Age | 57 ± 14 | 21 | 75 | 26% |
Weight (Kg) | 82 ± 13 | 60 | 110 | 16% |
Height (cm) | 171 ± 6 | 157 | 185 | 4% |
Body mass index (kg/m2) | 28 ± 5 | 21 | 38 | 16% |
Thorax (cm) | 107 ± 13 | 84 | 134 | 13% |
Abdomen (cm) | 107 ± 12 | 86 | 127 | 12% |
Bilirubin (mg/dL) | 0.8 ± 0.7 | 0.1 | 2.8 | 81% |
Creatinine (mg/dL) | 1.0 ± 0.3 | 0.4 | 1.6 | 32% |
Glucose (mg/dL) | 185 ± 70 | 84 | 340 | 39% |
AST (U/L) | 38 ± 20 | 18 | 103 | 55% |
ALP (U/L) | 32 ± 22 | 13 | 94 | 72% |
Hemoglobin (g/dL) | 11 ± 3 | 4 | 15 | 26% |
Urea (mg/dL) | 47 ± 17 | 25 | 77 | 36% |
K + (mEq/L) | 3.9 ± 0.5 | 3.3 | 5.1 | 12% |
Na + (mEq/L) | 150 ± 13 | 136 | 192 | 9% |
QUICK index | 81 ± 17 | 40 | 100 | 22% |
Mean ± SD or nº of Cases (%) | ||
---|---|---|
Age (years) | 51 ± 11 | |
Sex: Male—Female | 11 (48%)–12 (52%) | |
Body mass index (kg/m2) | 31 ± 6 | |
Glucose (mg/dL) | 116 ± 40 | |
TG (mg/dL) | 154 ± 67 | |
Total cholesterol (mg/dL) | 185 ± 27 | |
HDL-cholesterol (mg/dL) | 45 ± 16 | |
LDL-cholesterol (mg/dL) | 106 ± 29 | |
Total bilirubin (mg/dL) | 0.6 ± 0.3 | |
Albumin (g/dL) | 4.6 ± 0.2 | |
Platelets (103/µL) | 276 ± 90 | |
ALT (IU/L) | 50 ± 33 | |
AST (IU/L) | 43 ± 33 | |
ɣ-GT (IU/L) | 84 ± 61 | |
ALP (IU/L) | 83 ± 35 | |
Prothrombin (s) | 14 ± 2 | |
Hemoglobin (g/dL) | 14 ± 1 | |
Transferrin saturation (%) | 27 ± 14 | |
Insulin (µU/mL) | 22 ± 15 | |
Steatosis (%) | ||
Grade 0 | 6 (26%) | |
Grade 1 | 6 (26%) | |
Grade 2 | 6 (26%) | |
Grade 3 | 5 (22%) | |
Ballooning (%) | None (0) | 11 (48%) |
Moderate (1) | 9 (39%) | |
Severe (2) | 3 (13%) | |
Lobular inflammation (%) | None (0) | 8 (35%) |
Moderate (1) | 13 (56%) | |
Severe (2) | 2 (9%) | |
Fibrosis (%) | Stage 0 | 12 (53%) |
Stage 1 | 7 (30%) | |
Stage 2 | 1 (4%) | |
Stage 3 | 3 (13%) | |
Stage 4 | 0 (0%) | |
NAS scores (%) | NAS: 0–2 | 12 (53%) |
NAS: 3–4 | 4 (17%) | |
NAS: 5–8 | 7 (30%) | |
SAF activity scores (%) | A: 0–1 | 13 (57%) |
A: 2–3–4 | 10 (43%) |
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Quintás, G.; Caiment, F.; Rienda, I.; Pérez-Rojas, J.; Pareja, E.; Castell, J.V.; Jover, R. Quantitative Prediction of Steatosis in Patients with Non-Alcoholic Fatty Liver by Means of Hepatic MicroRNAs Present in Serum and Correlating with Hepatic Fat. Int. J. Mol. Sci. 2022, 23, 9298. https://doi.org/10.3390/ijms23169298
Quintás G, Caiment F, Rienda I, Pérez-Rojas J, Pareja E, Castell JV, Jover R. Quantitative Prediction of Steatosis in Patients with Non-Alcoholic Fatty Liver by Means of Hepatic MicroRNAs Present in Serum and Correlating with Hepatic Fat. International Journal of Molecular Sciences. 2022; 23(16):9298. https://doi.org/10.3390/ijms23169298
Chicago/Turabian StyleQuintás, Guillermo, Florian Caiment, Iván Rienda, Judith Pérez-Rojas, Eugenia Pareja, José V. Castell, and Ramiro Jover. 2022. "Quantitative Prediction of Steatosis in Patients with Non-Alcoholic Fatty Liver by Means of Hepatic MicroRNAs Present in Serum and Correlating with Hepatic Fat" International Journal of Molecular Sciences 23, no. 16: 9298. https://doi.org/10.3390/ijms23169298