Expression Levels of miR-30c and miR-186 in Adult Patients with Membranous Glomerulonephritis and Focal Segmental Glomerulosclerosis

Figures & data

Table 1 Demographic and Baseline Clinical Data

Characteristics	Total NS Patients	MGN	FSGS Group	P-value^a
No. of cases	60	30	30
Age, mean ± SD (years)	45.76±13	46.6± 13	44.5± 12.8	0.404
BMI (kg/m^2)	27.3±3.4	27.4± 3.6	27.3± 3.4	0.49
GFR(mL/min/1.73m)	80 (10–137)	80 (18–137)	79 (10–130)	0.324^b
Proteinuria (mg/24 h)	2497 (85–15,900)	3013 (85–15,000)	1360 (97–15,900)	0.26^b
Urea (mg/dL)	40.7± 19.4	38.2±16.5	44.2± 20	0.18
Uric acid (mg/dL)	6.5± 1.1	6.61± 1.2	6.3± 1.1	0.94
Serum creatinine (mg/dL)	1.2 ±0.5	2.21±0.55	1.2±0.5	0.17
Urine creatinine (mg/24h)	1124.38±407	1159.21 ±328	1074.63±500	0.742

Notes: GFR: estimated glomerular filtration rate as measured by GFR [186 × (Creatinine/88.4)^−1.154 × (Age)^−0.203 × (0.742 if female)] formula. The quantity data are expressed as mean ± SD. ^aMGN versus FSGS. ^bMedian (Min-Max) is presented. P-value is based on Mann–Whitney U-test.

Figure 1 The expression of miRNA-30c-5p and miRNA-186-5p in plasma and PBMCs. The expression of miRNA-30c-5p in (A) Plasma and (B) PBMCs between the studied groups. The expression of miRNA-186-5p in (C) Plasma and (D) PBMCs between the studied groups. The relative expression (fold change) was calculated by the 2^−ΔΔCt method. * and ○ are outliers.

Abbreviations: NS, nephrotic syndrome; FSGS, focal segmental glomerulosclerosis; MGN, membranous glomerulonephritis.

Table 2 Internal Correlations Between miRNA Levels in Plasma and Peripheral Blood Cells

	Plasma miR-30c	Plasma miR-186	PBMCs miR-30c	PBMCs miR-186
Plasma miR-30c	1	r= 0.779*	r= −0.191	r= −0.099
		p<0.001	p= 0.137	p= 0.449
Plasma miR-186		1	r= −0.176	r=−0.141
			p= 0.167	p= 0.273
PBMCs miR-30c			1	r= 0.60*
				p<0.001

Note: *Correlation is significant.

Abbreviation: r, correlation coefficient.

Table 3 Results of the ROC Curve Analysis for Discriminating Potential of PBMC and Plasma miR-30c and miR-186 Levels Among Pairs of the Studied Groups

	AUC	95% CI	P-value	Sensitivity	Specificity	Youden Index J	Associated Criterion
NS and controls
Plasma FC30c	0.50	0.418 to 0.585	0.97	87	30	0.13	>0.02
Plasma FC186	0.53	0.419 to 0.591	0.92	54	29	0.16	≤2.58
PBMC FC30c	0.65	0.565 to 0.728	0.001	46	96	0.4158	>2
PBMC FC186	0.45	0.418 to 0.582	1	0	100	0	>0
FSGS and MGN
Plasma FC30c	0.52	0.421 to 0.621	0.72	65	54	0.1849	>0.66
Plasma FC186	0.61	0.499 to 0.707	0.07	82	43	0.2472	>0.19
PBMC FC30c	0.54	0.437 to 0.647	0.48	89	30	0.1902	≤3.56
PBMC FC186	0.5	0.400 to 0.600	0.0	0	100	0	>0

Abbreviations: AUC, area under the curve; CI, confidence interval; ROC, receiver operating characteristic; NS, nephrotic syndrome; MGN, membranous glomerulonephritis; FSGS, focal segmental glomerulosclerosis; FC, fold change; PFC, plasma FC.

Figure 2 The combined ROC curves analysis of miR-30c and miR-186 in plasma and PBMCs samples. (A) Plasma and (B) PBMCs levels of the studied miRNAs could discriminate NS patients from controls. (C, D) ROC curve for the potential of the studied miRNAs in discriminating FSGS from MGN patients in (C) Plasma and (D) PBMCs. AUCs and the corresponding statistics of each miRNA between the groups are shown in Table 3.

Abbreviations: ROC, receiver operating characteristic; AUC, area under the curve.

Table 4 The Ten Top Enriched Biological Processes Among Target Genes of miR-30c and miR-186

GO ID	Biological Process	C	O	E	R	p-value^a
miR-30c
GO:0070671	Response to interleukin-12	49	9	1.518943743	5.925170068	1.67E^−05
GO:0040029	Regulation of gene expression, epigenetic	258	22	7.997703789	2.750789549	1.82E^−05
GO:0001763	Morphogenesis of a branching structure	196	18	6.075774971	2.962585034	3.96E^−05
GO:0002009	Morphogenesis of an epithelium	480	32	14.87944891	2.150617284	4.08E^−05
GO:0045023	G0 to G1 transition	45	8	1.394948335	5.734979424	6.30E^−05
GO:0031050	dsRNA processing	48	8	1.487,944,891	5.37654321	1.02E^−04
GO:0016569	Covalent chromatin modification	468	30	14.50746269	2.067901235	1.43E^−04
GO:0104004	Cellular response to environmental stimulus	320	23	9.919632606	2.318634259	1.67E^−04
GO:1,990,823	Response to leukemia inhibitory factor	95	11	2.94489093	3.735282651	1.69E^−04
GO:0010608	Posttranscriptional regulation of gene expression	486	30	15.06544202	1.9913123	2.74E^−04
miR-186
GO:0016071	mRNA metabolic process	765	79	30.80383	2.564616	8.88E^−15
GO:0010629	Negative regulation of gene expression	1733	127	69.78175	1.81996	1.01E^−11
GO:0006412	Translation	613	62	24.68333	2.511817	2.06E^−11
GO:0006396	RNA processing	887	78	35.71633	2.183875	5.11E^−11
GO:0043043	Peptide biosynthetic process	636	62	25.60946	2.420981	9.58E^−11
GO:0051276	Chromosome organization	1143	92	46.02454	1.998933	1.00E^−10
GO:0044265	Cellular macromolecule catabolic process	1108	88	44.61522	1.972421	5.32E^−10
GO:0008380	RNA splicing	417	46	16.79111	2.739545	5.62E^−10
GO:0043604	Amide biosynthetic process	766	68	30.8441	2.204636	6.64E^−10
GO:0006397	mRNA processing	487	50	19.60976	2.549751	1.20E^−09

Note: ^aThe Benjamini–Hochberg adjusted p-value.

Abbreviations: C, total number of genes in the category; O, number of observed target genes in the category; E, number of expected genes in the category; R, fold enrichment.