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. 2020 Apr 11;10(4):589. doi: 10.3390/biom10040589

Table 1.

Circulating microRNAs (miRNAs) in pre- and postmenopausal osteoporosis (PM OP).

Study Population Variables in Pre-Analytical Phase:
- Sample matrix
- Centrifugation conditions
- Storage conditions		 Quantification Method
- Target miRNAs
- Analytical method
- Applied normalization strategy		 - Validated miRNAs
- AUC; sensitivity; specificity		 Reference
Screening Validation
PM Chinese women with normal (n = 40), osteopenia (n = 40) or OP (n = 40) range of BMD - Plasma
- n.d.
- Liquid nitrogen		 - miR-146a, miR-133a, and miR-21
- RT-qPCR
- miR-16		 - ↑ miR-133a and ↓ miR-21 in OP and osteopenia vs. control
- n.d.		 [72]
Screening: PM Chinese women with OP (n = 25) or with osteopenia (n = 23) Validation: PM Chinese women with OP (n = 32), with osteopenia (n = 30), and with normal range of BMD (n = 24) - Whole blood, treated using RBC Lysis Solution;
- 450g, 10 min
- n.d.		 - miR-660-5p, miR-590-5p, miR-194-5p, miR-151a-3p, miR-151b, and miR-130b-3p
- RT-qPCR (comprehensive miRNA expression analysis by Agilent Human miRNA microarray)
- snRNU6		 - miR-194-5p
- RT-qPCR
- snRNU6		 Screening:
- ↑ miR-590-5p, miR-194-5p, miR-151b, miR-151a-3p, and miR-130b-3p in OP vs. osteopenia
- n.d.
Validation:
- ↑ miR-194-5p in OP and osteopenia vs. control
- n.d.		 [75]
Screening: PM Chinese women with OP (n = 5) and healthy premenopausal women (n = 5)
Validation: PM Chinese women with OP (n = 81) and healthy premenopausal women (n = 74) 		-Serum
- n.d.
- n.d. 		- 851 miRNAs
- Agilent Human miRNA Microarray
- n.d.		 - miR-27a
- RT-qPCR
- snRNU6		 - ↓ miR-27a in OP vs. control
- n.d.		 [76]
PM women divided in controls (n = 57) and OP (n = 17) based on lumbar spine/femoral neck/total hip T-score ≤−2.5 SD - EDTA-Plasma
- (i) 10 min, 2800 rpm, 4 °C; (ii) 15 min, 9600g, 4 °C
- −80 °C		 - miR-574-5p, miR-423-5p, miR-199a-3p, miR-148a-3p, miR-126-3p, miR-30d-5p, miR-30e-5p, miR-7d-5p, and miR-7e-5p
- RT-qPCR
- Combination of miR-16-5p and let-7a-5p		 - ↑ miR-148a-3p in OP vs. control
- n.d.		 [77]
PM women with OP (n = 10), osteopenia (n = 7), or normal (n = 19) range of BMD - Serum
- (i) 10 min, 2000g, 4 °C; (ii) 20 min, 12,000g, 4 °C
- −80 °C		 - miR-425-5p, miR-361-5p, miR-345-5p, miR-215, miR-191a-3p, miR-185-5p, miR-142-3p, miR-103-3p, miR-30a-5p, miR-30b-5p, miR-30e-5p, miR-29b-3p, and miR-26a-5p
- RT-qPCR
- miR-25-3p		 - ↓ miR-30b-5p in both osteopenia and OP vs. control;
↓ miR-328-3p, miR-142-3p, miR-103-3p in OP vs. HC
- 0.87; 80%; 100% (miR-328-3p),
0.79; 70%; 79% (miR-142-3p),
0.80; 80%; 72% (miR-103-3p),
0.79; 70.6%; 79.0% (miR-30b-5p)		 [78]
Screening: PM Mexican-Mestizo women with normal (n = 20) and OP (n = 20) hip BMD Validation: PM Mexican-Mestizo women with normal (n = 22), OP (n = 26), and osteopenia (n = 28) hip BMD, and with fractures (n = 21) - Serum, obtained within 1h of collection
- n.d.
- −80 °C		 - 754 miRNAs
- TaqMan Array Human MicroRNA A+B Cards Set v3.0
- snRNU6		 - miR-885-5p, miR-140-3p, and miR-23b-3p
- RT-qPCR
- snRNU6		 - ↑ miR-885-5p for osteopenia vs. HC;
↑ miR-140-3p and miR-23b-3p for osteopenia, OP, and fracture vs. HC
- 0.69 (miR-885-5p),
0.84 for osteopenia, 0.96 for OP, 0.92 for fracture (miR-140-3p),
0.73 for osteopenia, 0.69 for OP, 0.88 for fracture (miR-23b-3p) 		[79]
PM Chinese women: HC (n = 15) and OP (n = 15) - Serum
- n.d.
- n.d. 		- miR-3065-5p and miR-338-3p
- RT-qPCR
- cel-miR-39-3p		 - ↑ miR-3065-5p and miR-338-3p in OP vs. HC
- 0.87 (miR-3065-5p)
0.74 (miR-338-3p)		 [80]
PM OP women (n = 36) and non-OP (n = 36) - Serum
- n.d.
- n.d.		 - miR-483-5p
- RT-qPCR
- U6		 - ↑ miR-483-5p in OP vs. non-OP
- n.d.		 [81]
PM OP treated with teriparatide (n = 30) or denosumab (n = 30) for 12 months. - Serum
- n.d.
- −80 °C 		- miR-2861, miR-503, miR-422a, miR-335-5p, miR-222-5p, miR-218-5p, miR-135b, miR-133a, miR-124-3p, miR-33-3p, miR-29c-3p, miR-27a, miR-26a-5p, miR-24-2-5p, miR-23a-3p, and miR-21-5p
- RT-qPCR
- Combination of SNORD95, SNORD96A,
RNU6-2		 - ↓ miR-33-3p after 3 months of teriparatide treatment;
↓ miR-133a after 12 months of teriparatide treatment
- n.d. 		[83]
PM women divided into: osteopenia with (n = 26) or without anti-OP therapy (n = 14), OP with (n = 29) or without (n = 17) anti-OP therapy; healthy premenopausal women (HC, n = 14) - Serum, clotted at RT for 1 h
- 1000g, 10 min
- −80 °C		 - miR-1290, miR-497-5p, miR-204-3p, miR-181c-5p
- RT-qPCR
- 5S		 - ↓ miR-497-5p, miR-204-3p in osteopenia vs. HC;
↓ miR-497-5p, miR-204-3p, miR-181c-5p in OP vs. HC;
↑ miR-497-5p, miR-181c-5p in both OP and osteopenia with anti-OP therapy vs. OP and osteopenia without anti-OP therapy
- n.d.		 [84]
PM women divided into: OP (n = 35) and non-OP (n = 35);
Premenopausal women divided in: OP (n = 35) and non-OP (n = 35).		 - Serum
- 3000g, 15 min
- −80 °C		 - miR-637, miR-208a-3p, miR-155–5p
- RT-qPCR
- Hs_Snord68_11		 - ↑ miR-208a-3p and ↓ miR-155-5p in premenopausal OP vs. premenopausal non-OP;
↑ miR-637, miR-208a-3p, miR-155-5p in PM OP vs. PM non-OP;
↑ miR-637, miR-208a-3p, miR-155-5p in PM OP vs. premenopausal OP
- 0.82;77.1%;82.9% (miR-208a-3p) and 0.90;94.3%;77.1% (miR-155-5p) for premenopausal OP vs. premenopausal non-OP;
0.81;77.1%;85.7% (miR-637), 0.85;80.0%;82.9% (miR-208a-3p), and 0.83;80.0%;80.0% (miR-155-5p) for PM OP vs. PM non-OP;		 [85]

n.d.: not determined; AUC: Area Under the Curve; BMD: Bone Mineral Density; HC: Healthy Controls; OP: Osteoporosis; PM: Postmenopausal; RT: Room Temperature; RT-qPCR: Real-time quantitative Polymerase Chain Reaction. ↑: increased levels of the validated circulating miRNA; ↓: decreased levels of the validated circulating miRNA.