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| Status |
Public on Dec 30, 2022 |
| Title |
Extracellular vesicle microRNA profiling in synovial fluids and plasma from Oligoarticular Juvenile Idiopathic Arthritis patients: methodological challenges and identification of putative disease biomarkers |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by RT-PCR
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| Summary |
New early low-invasive biomarkers are demanded for the management of Oligoarticular Juvenile Idiopathic Arthritis (OJIA), the most common chronic inflammatory arthritis in childhood and a leading cause of disability. Extracellular vesicles (EVs) released in the synovial fluid (SF) of inflamed joints have gained increased recognition as mediators of arthritis pathogenesis and as a source of biomarkers. No standardized protocol is available, yet, for the analysis of EV-derived microRNAs (EV-miRs) in human SF specimens, and their potential as biomarkers in OJIA has not been explored. This study proposes an optimized procedure for profiling EV-miRs in SF and represents the first investigation of the miRNome in EV isolated from OJIA patients at disease onset. We present data showing efficient recovery of EVs from SF aspirates and optimal profiling of their miRNA content. We identified a subset of 15 EV-miRs whose expression was deregulated in SF. Analysis of EV-miRs target genes evidenced the enrichemnt of processes related to inflammation, cartilage/bone homeostasis, hypoxia, and hormone metabolism, suggesting their contribution to disease pathogenesis and potential as new putative biomarkers. In addition, we detected altered expression levels of several circulating EV-miRs able to discriminate new-onset OJIA patients from control children. Among them, 16 were consensually overexpressed in both PL and SF from OJIA patients respect to controls, potentially representing a disease-associated molecular “fingerprint” measurable at both the systemic and local levels with high diagnostic potential. These results improve our understanding of OJIA molecular mechanisms and provide an important contribution in the search of new candidate biomarkers for early OJIA diagnosis and the design of novel tailored therapies.
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| Overall design |
Thirteen patients newly diagnosed with OJIA at the Pediatric and Rheumatologic Department of the Gaslini Institute, Genova, Italy on the basis of the 2001 International League of Associations for Rheumatology (ILAR) classification criteria 1 were enrolled consecutively in the study from october 2018 through February 2020 (Table I). All patients had clinically active disease, with joint effusion, swelling, pain, and stiffness, and underwent arthrocentesis and intra-articular steroid administration as part of routine clinical care. Arthrocentesis was performed with ultrasound guidance under local anesthesia or, in case of younger patients or multiple joints, under general anesthesia. Patients who were having anti-inflammatory treatment (steroids, disease-modifying anti-rheumatic drugs-DMARDs, or biological therapy) at the time of arthrocentesis were excluded from the study . Eight age- and gender-matched children undergoing minor orthopedic procedures at the Gaslini Institute were enrolled as control group. Detailed clinical and laboratory examination of control subjects was carried out to rule out infections, inflammatory, and chronic diseases. The protocol of the study was reviewed and approved by the Ethical Committee of the Liguria Region (Approval 165/2018), and the procedures were carried out according to the approved guidelines and in adherence to the general ethical principles set forth in the Declaration of Helsinki. Written informed consent to participate to the study was obtained from the parents or the patient legal guardian prior to sample collection.
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| Web link |
https://pubmed.ncbi.nlm.nih.gov/36178089/
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| Contributor(s) |
Raggi F, Cangelosi D, Bosco MC |
| Citation(s) |
36178089 |
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| Submission date |
Dec 20, 2021 |
| Last update date |
Mar 31, 2023 |
| Contact name |
Federica Raggi |
| E-mail(s) |
federicaraggi@gaslini.org
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| Organization name |
IRCCS Istituto Giannina Gaslini
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| Lab |
Molecular biology
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| Street address |
via genrolamo gaslini, 5
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| City |
Genova |
| ZIP/Postal code |
16147 |
| Country |
Italy |
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| Platforms (1) |
| GPL19079 |
Applied Biosystems (Thermo Fisher) TaqMan Human microRNA Array (RT-PCR miRNA Profiling) |
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| Samples (24)
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| Relations |
| BioProject |
PRJNA790988 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE191326_non-normalized_data.txt.gz |
19.3 Kb |
(ftp)(http) |
TXT |
| GSE191326_normalized_data.txt.gz |
20.8 Kb |
(ftp)(http) |
TXT |
| Processed data are available on Series record |
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