Differentially Expressed Circulating microRNAs in the Development of Acute Diabetic Charcot Foot

Figures & data

Table 1.  Principal characteristics of the patients included in the study. Diabetes but no neuropathy (G3), diabetes with neuropathy (G2) and diabetes with both neuropathy and Charcot foot (G1). Data are represented as mean (SD) or number (%).

Characteristics	G1: Diabetes with Charcot, n = 17	G2: Diabetes with neuropathy, n = 17	G3: Diabetes without neuropathy, n = 17	p-value
Age, years	55.76 (9.53)	54.35 (9.21)	53.76 (9.21)	0.81
Gender (male/female)	11/6	12/5	12/5	0.75
BMI, kg/m^2	31.33 (5.86)	34.34 (7.12)	29.93 (3.84)	0.08
HbA1c, %	8.1% (1.80)	8.36% (2.12)	8.24% (1.90)	0.92
Fasting glucose, mmol/L	9.33 (4.51)	10.69 (7.01)	10.28 (4.31)	0.75
Diabetes duration, years	15.23 (5.50)	15 (7.41)	8.47 (6.42)	0.0049
Systolic BP, mmHg	135.64 (18.53)	135.64 (16.57)	131.5 (15.95)	0.74
Diastolic BP, mmHg	73.29 (6.97)	74.36 (10.04)	77.06 (9.21)	0.48
eGFR, ml/min	82.81 (34.38)	73.59 (46.30)	94.69 (19.42)	0.22

BP: Blood pressure; eGFR: Estimated glomerular filtration rate.

Figure 1.  Volcano plots.

Volcano plots showing differentially expressed miRNAs in the following comparisons: T2D with Charcot foot over T2D with neuropathy (Group 1 vs Group 2) (A); T2D with Charcot foot over T2D without neuropathy (Group 1 vs Group 3) (B); and T2D with neuropathy over T2D without neuropathy (Group 2 vs Group 3) (C). miRNAs with more than twofold change and a significant p-value after applying Bonferroni correction are shown in green. Those, which show more than twofold change but whose p-value significance is achieved by discounting Bonferroni correction are shown in yellow. Gray dots represent miRNAs exhibiting less than twofold change, while red dots stand for those showing more than twofold change but lacking statistical significance. Solid vertical lines indicate twofold upregulation or downregulation on the log scale. Dashed horizontal lines either mark p-value = 0.05 on the log scale (red line), or Bonferroni-corrected p-value cut-off (green line).

T2D: Type 2 diabetes.

Figure 2.  Venn diagram.

A three-way Venn diagram representing the apparition of miRNAs in each comparison. The one in the center (black) appears in every contrast, the one at the merging of two circles (orange, purple, green) appears in two comparisons and the one in once circle only (yellow, red, blue) appears in only one set of data.

Figure 3.  A network of predicted genes affected by differentially expressed miRNA from G1 versus G2 comparison created by Ingenuity Pathway Analysis.

The networks hypothesized by Ingenuity Pathway Analysis are based on the molecular relationships, interactions and pathway associations between the methylated candidate genes as shown in a graphical representation.

Figure 4.  Role of miRNA differentially expressed between G1 versus G2 in the osteoclast differentiation.

The graphical representation had been generated by Ingenuity Pathway Analysis and reflects the molecular relationships between miRNAs and candidate genes. miRNAs in red were overexpressed in Charcot foot (CF) patients as compared with Type 2 diabetes (T2D) patients with neuropathy, while the ones in green were downexpressed in CF patients compared with T2D patients with neuropathy.

Table 2.  miRNAs differentially expressed between G1 and G2 and involved in the osteoclast differentiation.

The color corresponds to the group defined within the Venn diagram (Figure 2). The three miRNAs inside the bold square are the one that passed the Bonferroni significance.