Analysis of microRNAs in Small Urinary Extracellular Vesicles and Their Potential Roles in Pathogenesis of Renal ANCA-Associated Vasculitis
Abstract
:1. Introduction
2. Results
2.1. Next Generation Sequencing Based Phase
2.2. RT-PCR Based Confirmation Phase
2.3. Comparison with Proteomic Results and Bioinformatic Analysis
3. Discussion
4. Materials and Methods
4.1. Patients and Urine Sample Collection
4.2. Urine Sample Preparation
4.3. Urinary Extracellular Vesicle Enrichment and miRNA Isolation
4.4. Next Generation Sequencing Based Phase
4.5. Single Target RT-PCR Based Confirmation Phase
4.6. Bioinformatic Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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miRNA | p-Value | FC (P/C) | SD (Controls) | SD (Patients) | Power |
---|---|---|---|---|---|
miR-26a-5p | 5.982 × 10−6 | 0.422 | 0.276 | 0.053 | 1.0000 |
miR-192-5p | 5.417 × 10−4 | 0.312 | 0.546 | 0.099 | 1.0000 |
miR-191-5p | 3.871 × 10−2 | 0.720 | 0.182 | 0.077 | 1.0000 |
miR-31-3p | 2.439 × 10−4 | 2.933 | 0.166 | 0.232 | 1.0000 |
miR-106b-5p | 4.253 × 10−4 | 2.841 | 0.124 | 0.214 | 1.0000 |
miR-99a-5p | 4.257 × 10−3 | 4.329 | 0.161 | 0.282 | 1.0000 |
miR-30a-5p | 9.190 × 10−3 | 2.551 | 0.154 | 0.172 | 1.0000 |
miR-182-5p | 1.348× 10−2 | 3.175 | 0.190 | 0.171 | 1.0000 |
miR-24-3p | 2.075 × 10−2 | 1.412 | 0.071 | 0.073 | 1.0000 |
miR-200a-3p | 3.871 × 10−2 | 1.422 | 0.095 | 0.171 | 0.9999 |
Parameters | NGS Initial Phase | RT-PCR Confirmation Phase | ||
---|---|---|---|---|
AVV Patients n = 10 | Healthy Controls n = 10 | AVV Patients n = 24 | Healthy Controls n = 16 | |
Sex (Male/Female) | 6/4 | 6/4 | 12/12 | 8/8 |
Age (median, range, years) | 65.5 (37–74) | 55 (42–74) | 63.5 (21–78) | 57 (42–74) |
S-creatinine (median, range, μmol/L) | 241.5 (113.1–438.3) | 79.1 (64.4–86.1) | 302.0 (52.3–480.1) | 82.2 (64.4–91.3) |
Proteinuria (median, range, g/24 h) | 1.62 (0.21–3.60) | 0.05 (0.03–0.13) | 1.44 (0.11–3.60) | 0.05 (0.03–0.13) |
Hemoglobin (median, range, g/L) | 91.0 (79–142) | n/a | 98.5 (77–142) | n/a |
C-reactive protein (median, range, mg/L) | 103.5 (1.1–153.0) | n/a | 12.0 (1.1–153.0) | n/a |
Organ involvement (kidney/lung/ENT/eye/skin) | 10/3/0/1/0 | n/a | 24/9/4/1/2 | n/a |
ANCA-subtypes (PR3/MPO/neg.) | 3/7 | n/a | 7/15/2 | n/a |
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Frydlova, J.; Zednikova, I.; Satrapova, V.; Pazourkova, E.; Santorova, S.; Hruskova, Z.; Tesar, V.; Vokurka, M.; Prikryl, P.; Korabecna, M. Analysis of microRNAs in Small Urinary Extracellular Vesicles and Their Potential Roles in Pathogenesis of Renal ANCA-Associated Vasculitis. Int. J. Mol. Sci. 2022, 23, 4344. https://doi.org/10.3390/ijms23084344
Frydlova J, Zednikova I, Satrapova V, Pazourkova E, Santorova S, Hruskova Z, Tesar V, Vokurka M, Prikryl P, Korabecna M. Analysis of microRNAs in Small Urinary Extracellular Vesicles and Their Potential Roles in Pathogenesis of Renal ANCA-Associated Vasculitis. International Journal of Molecular Sciences. 2022; 23(8):4344. https://doi.org/10.3390/ijms23084344
Chicago/Turabian StyleFrydlova, Jana, Iveta Zednikova, Veronika Satrapova, Eva Pazourkova, Sarka Santorova, Zdenka Hruskova, Vladimir Tesar, Martin Vokurka, Petr Prikryl, and Marie Korabecna. 2022. "Analysis of microRNAs in Small Urinary Extracellular Vesicles and Their Potential Roles in Pathogenesis of Renal ANCA-Associated Vasculitis" International Journal of Molecular Sciences 23, no. 8: 4344. https://doi.org/10.3390/ijms23084344