The Profiling of 179 miRNA Expression in Serum from Limb Girdle Muscular Dystrophy Patients and Healthy Controls
Abstract
:1. Introduction
2. Results
2.1. Clinical Aspects and Muscle Biopsy
2.2. Data Control Quality
2.3. The Expression Analysis of Serum-Circulating miRNAs
2.4. The Identification of miRNAs Differentially Expressed in Each LGMDR Pool and the Functional Enrichment of miRNA Targets
2.5. Validation and Specificity Evaluation of miRNA Signature Identification in LGMD Patients
2.6. The Correlation of Circulating miRNAs with Demographic, Morphological and Functional Parameters
2.7. The Specificity of the miRNA Signature
3. Discussion
Study Limitation
4. Materials and Methods
4.1. Patients
4.2. Ethics Statement
4.3. Morphology
4.4. Serum Collection
4.5. RNA Isolation
4.6. cDNA Synthesis
4.7. RT-PCR Based miRNA Assays (Real Time PCR)
4.8. Real-Time Quantitative PCR (RT-qPCR)
4.9. Data Control Quality
4.10. Bioinformatics Tools
4.11. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Patients | LGMDR1 | LGMDR2 | LGMDR3 | LGMDR4 | Ctrl |
---|---|---|---|---|---|
(N = 4) | (N = 4) | (N = 4) | (N = 4) | (N = 8) | |
Sex | 1F/3M | 3F/1M | 3F/1M | 1F/3M | 4F/4M |
Age | 47.25 ± 15.51 | 38.25 ± 14.52 | 46.75 ± 16.25 | 19.50 ± 6.61 | 37.70 ± 0.63 |
Age at onset (yrs) | 30.75 ± 7.88 | 20.75 ± 12.33 | 12.50 ± 5.44 | 7.25 ± 4.03 | NA |
Disease duration (yrs) | 16.5 ± 10.37 | 17.25 ± 5.85 | 32.50 ± 17.74 | 9.0 ± 8.36 | NA |
Morphology | |||||
Fibrosis (% area) | 20.44 ± 2.23 p < 0.0001 | 13.24 ± 0.45 p < 0.0001 | 9.49 ± 0.48 p = 0.8619 | 13.90 ± 2.22 p = 0.0002 | 9.39 ± 0.29 |
Centronucleated Fiber | 2.53 ± 0.66 p < 0.0001 | 3.40 ± 0.61 p < 0.0001 | 1.53 ± 0.43 p = 0.0081 | 0.80 ± 1.03 p = 0.320 | 0.48 ± 0.16 |
CSA type I (mm2) | 2657 ± 116 p < 0.0001 | 2451 ± 89 p < 0.0001 | 3920 ± 114 p = 0.0145 | 1893 ± 67 p < 0.0001 | 4215 ± 38 |
CSA type II (mm2) | 2227 ± 72 p < 0.0001 | 2118 ± 69 p < 0.0001 | 3563 ± 80 p = 0.0019 | 2158 ± 58 p < 0.0001 | 3819 ± 36 |
Clinical data | |||||
CK (IU/L) | 1813 ± 550 | 931 ± 185 | 1996 ± 1457 | 6151 ± 1958 | 60-190 |
Loss of ambulation | 1/4 | 1/4 | 1/4 | 2/4 | NA |
Functional tests | Normal values | ||||
MFM Dimension 1 (score) | 14.75 ± 15.79 | 23.25 ± 9.84 | 21.75 ± 16.45 | 17.25 ± 20.25 | 39 |
MFM Dimension 2 (score) | 30.00 ± 5.16 | 30.00 ± 14.69 | 32.00 ± 6.73 | 19.00 ± 19.69 | 36 |
MFM Dimension 3 (score) | 18.75 ± 1.79 | 18.25 ± 9.33 | 20.00 ± 1.15 | 13.00 ± 8.90 | 21 |
MFM tot (score) | 63.50 ± 21.79 | 71.5 ± 30.50 | 73.75 ± 23.76 | 49.25 ± 48.50 | 96 |
Six Minute Walking test (6-MWT) (meter) | 284.3 ± 141.8 (n = 3) | 381.0 ± 152.0 (n = 3) | 461.2 ± 168.8 (n = 3) | 530.5 ± 67.1 (n = 2) | 615.5 ± 31.8 (n = 102) [22] |
NSAD | 21.00 ± 22.68 | 32.25 ± 15.41 | 31.00 ± 22.13 | 23.00 ± 27.00 | 54 |
PUL | 27.25 ± 10.14 | 35.00 ± 15.15 | 34.00 ± 10.09 | 22.50 ± 20.85 | 42 |
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Magri, F.; Napoli, L.; Ripolone, M.; Ciscato, P.; Moggio, M.; Corti, S.; Comi, G.P.; Sciacco, M.; Zanotti, S. The Profiling of 179 miRNA Expression in Serum from Limb Girdle Muscular Dystrophy Patients and Healthy Controls. Int. J. Mol. Sci. 2023, 24, 17402. https://doi.org/10.3390/ijms242417402
Magri F, Napoli L, Ripolone M, Ciscato P, Moggio M, Corti S, Comi GP, Sciacco M, Zanotti S. The Profiling of 179 miRNA Expression in Serum from Limb Girdle Muscular Dystrophy Patients and Healthy Controls. International Journal of Molecular Sciences. 2023; 24(24):17402. https://doi.org/10.3390/ijms242417402
Chicago/Turabian StyleMagri, Francesca, Laura Napoli, Michela Ripolone, Patrizia Ciscato, Maurizio Moggio, Stefania Corti, Giacomo Pietro Comi, Monica Sciacco, and Simona Zanotti. 2023. "The Profiling of 179 miRNA Expression in Serum from Limb Girdle Muscular Dystrophy Patients and Healthy Controls" International Journal of Molecular Sciences 24, no. 24: 17402. https://doi.org/10.3390/ijms242417402