Table 3.
miRNAs associated with other types of OP and related fracture risk.
| Study | Study design | Biomarker source | Sample handling | Quantification platform | Evaluated miRNA | Normalization strategy | Validated miRNA biomarker | Potential target gene | AUC-Sensitivity(%)-Specificity(%) | Limits |
|---|---|---|---|---|---|---|---|---|---|---|
| (Chen et al., 2014b) | 4 Chinese OP patients (3 women and 1 man, age 76-88 years) and 5 Chinese subjects with normal BMD (2 women and 3 men, age 19-44 years) | BM-MCSs | Bone marrow aspirated from iliac crest and used for BM-MCSs isolation | SYBR Green RT-qPCR | miR-125b | snRNU6 | ↑miR-125b in OP group vs. non-OP group | OSX (Identified using TargetScan and PicTar database, and validated by in vitro experiments) and RUNX2 for miR-125b | / | Small sample size; no information about the stem-loop arm of miRNA origin; no ROC analysis |
| (Weilner et al., 2016) | 14 men (mean age ∼53 years) with idiopathic osteoporosis and 11 age-matched HC | Cell-free plasma and plasma microvesicles | Filtration and differential centrifugation methodologies for microvescicle purification | TaqMan RT-qPCR | miR-31 | snRNU6 | ↑miR-31 in OP group vs. HC | FZD3 (validated by in vitro experiments for miR-31) | / | Small sample size; no information about the stem-loop arm of miRNA origin; study mainly focused on miRNA evaluation by in vitro studies; no ROC analysis |
| (Wang et al., 2013c) | 40 Chinese patients with fracture (age 60-90 years) and 9 Chinese HC (control group) | Bone specimens | Femurs collected during surgery | RT-PCR | Not specified | Not specified in this paper | ↑miR-214a in older individuals | ATF4 (identified for miR-214a using miRBase and validated by in vitro-in vivo experiments) | / | Small sample size of the HC group; confusing information about the comparisons done among groups; evaluated miRNAs and data normalization not explained in this paper; no information about the stem-loop arm of miRNA origin; no ROC analysis |
| (Kelch et al., 2017) | 28 patients with hip fracture: 7 men + 7 women with OP and 7 men + 7 women with AO (control group) | Cell-free serum and bone tissue | Blood collected 2 h post-fracture (OP) or pre-operation (non-OP) into S-Monovette polypropylene tubes, placed for 30 min at RT upright, centrifuged for 10 min at 1900g, serum stored at -80°CFemoral head samples collected during surgery (within 8 h after fracture in OP group). Cylindrical bone samples obtained from middle of each femoral head, cut into small pieces with Luer forceps, rinsed with D-PBS, collected in TRI-Reagent, snap frozen in liquid nitrogen, and mechanically ground. The bone powder collected with TRI-Reagent and stored at -80°C | miScript SYBR Green RT-qPCR | miR-21-5p, miR-23a-3p, miR-24-3p, miR-93-5p, miR-100-5p, miR-122-5p, miR-124-3p, miR-125b-5p, and miR-148a-3p | SNORD96a | ↑ miR-21-5p, miR-23a-3p, miR-24-3p, miR-93-5p, miR-100-5p, miR-122-5p, miR-124-3p, and miR-148a-3p in OP serum vs. control↑ miR-21-5p, miR-24-3p, miR-93-5p, miR-100-5p and miR-125b-5p in OP tissues vs. control↑ miR-21-5p, miR-23a-3p, miR-24-3p, miR-93-5p, miR-100-5p, and miR-125b-5p in OP osteoblasts vs. control↑ miR-21-5p, miR-93-5p, miR-100-5p, miR-122-5p, miR-124-3p, miR-125b-5p, and miR-148a-3p in OP osteoclasts vs. control | / | / | no evaluation of the target genes; no ROC analysis |
| (Kocijan et al., 2016) | Patients with idiopathic (16 men and 10 premenopausal women); HC without low-trauma fracture (16 men and 12 premenopausal women). | Cell-free serum | Fasting blood samples immediately centrifuged and serum stored a -80°C | SYBR Green RT-qPCR | 187 miRNAs tested | Global mean | ↑ miR-152-3p, miR-335-5p, miR-320a and↓ let-7b-5p, miR-7-5p, miR-16-5p, miR-19a-3p, miR-19b-3p, miR-29b-3p, miR-30e-5p, miR-93-5p, miR-140-5p, miR-215-5p, miR-186-5p, miR-324-3p, miR-365a-3p, miR-378a-5p, miR-532-5p, and miR-550a-3p in fractured groups vs. their control groups | / | 0.962 (miR-152-3p), 0.959 (miR-30e-5p), 0.950 (miR-324-3p), 0.947(miR-140-5p), 0.944 (miR-19b-3p), 0.939 (miR-335-5p), 0.929 (miR-19a-3p), 0.909 (miR-550a-3p), 0.898 (miR-186-5p), 0.898 (miR-532-5p), 0.872 (miR-378a-5p), 0.870 (miR-320a), 0.879 (miR-93-5p), 0.857 (miR-16-5p), 0.853 (miR-215-5p), 0.852 (let-7b-5p), 0.824 (miR-7-5p), 0.838 (miR-29b-3p), and 0.809 (miR-365a-3p) for fracture groups vs. control groups | No evaluation of the target genes; arbitrary choice of the screened miRNAs |
| (Mandourah et al., 2018) | 12 (1 male/11 females) non-OP controls, 61 (9 males/52 females) osteopenia without fracture, 15 (2 males/13 females) osteopenia with fracture, 33 (6 males/27 females) OP without fracture, and 18 (2 males/16 females) OP with fracture | Cell-free serum and plasma | Serum/plasma samples obtained by centrifuging at 2500g and RT for 30 min. Supernatants further centrifuged at 14000g and 4°C for 30 min. Samples stored at -80°C | Screening: Human Serum and Plasma miRNA PCR arrays Validation: miScript SYBR Green RT-qPCR |
Screening: 370 miRNAs tested Validation: 40 miRNAs tested |
SNORD96A and RNU6-6P | ↓ miR-122-5p and miR-4516 in OP vs. non-OP and osteopenia patients | BMP2K, FSHB, IGF1R, VDR, SPARC, TSC22D3 and RUNX2 (miR-122-5p and miR-4516); ANKH, ALPL, CNR2, CD44, LRP6, and ESR1 (miR-122-5p); AR and CNR1 (miR-4516) (identified using miRWalk2.0 database but not validated in the study) | 0.727-71-62 (miR-4516) and 0.752 (miR-122-5p+miR-4516) for OP | Small sample size; confusing information about the comparisons done among groups; no validation of the identified target genes |
| (Heilmeier et al., 2016) | 80 PM women; two study arms with two groups each:T2DM arm composed of T2DM women with (n = 20) and without (n = 20) fragility fractures since T2DM onsetOP arm composed of healthy non-T2DM PM women with OP fragility fracture (n = 20), and control group of fracture-free PM women (n = 20). | Cell-free serum | Fasting blood placed for 40 min upright and centrifuged for 15 min at 2000g. | SYBR Green Low-density qPCR platform | 375 miRNAs tested | Cq values computed using second derivative maximum method provided with LC480 II software. | Most abundant miRNAs among the top 10 four-miRNAs models:↓ miR-382-3p in T2DM and OP with fragility fracture vs. respective controls↑ miR-550a-5p and miR-96-5p in T2DM fragility fracture group vs. controls↓ miR-188-3p and miR-942 in OP fracture group vs. controls | / | 10 candidate four-miRNA models displayed AUC values (0.922 -0.965) for identifying fracture status in T2DM.10 candidate four-miRNA models displayed AUC values (0.972 -0.991) for identifying fracture status in OP group. | No evaluation of the target genes; arbitrary choice of the screened miRNAs |
ALPL, alkaline phosphatase; ANKH, ANKH inorganic pyrophosphate transport regulator; AR, androgen receptor; ATF4, activating transcription factor 4; BMD, bone mineral density; BM-MCSs, bone marrow mesenchymal stem cells; BMP2K, BMP2 inducible kinase; CNR2, cannabinoid receptor 2; ESR1, estrogen receptor 1; FSHB, follicle stimulating hormone subunit beta;FZD3, frizzled-3; HC, healthy controls; IGF1R, insulin-like growth factor 1 receptor;OA, osteoarthritis; OP, osteoporosis; OSX, osterix; RT-qPCR, real-time quantitative polymerase chain reaction; RUNX2, runt-related transcription factor 2; RUNX2, runt-related transcription factor 2; SPARC, secreted protein acidic and cysteine rich; T2DM, type 2 diabetes mellitus; TSC22D3, TSC22 domain family member 3; VDR, vitamin D receptor; WIF1, WNT inhibitory factor 1; WISP1, WNT1-inducible-signaling pathway protein 1.