MiRNA-based “fitness score” to assess the individual response to diet, metabolism, and exercise

Figures & data

Figure 1. Study design of the pilot study 2018 (A) and the intervention study 2019 − 2020 (B). T0, begin, T1, after 10-/12-weeks, T2, after 10-month, DBS, dried blood spot, BIA, bioelectrical impedance analysis, FFQ, food frequency questionnaire, HRM, high-resolution melting, SNP, single nucleotide polymorphism.

Table 1. Results of the analyzed markers of the participants of the sports intervention group. Results were given in mean ± SD.

	Total (n, 61)			Male (n, 20)		Female (n, 41)
	Fold Change ± SD	Paired t-test	ANCOVA	Fold Change ± SD	Paired t-test	Fold Change ± SD	Paired t-test
miR-19b-3p	1.212 ± 0.764	0.463	0.192	1.119 ± 0.619	0.641	1.221 ± 0,832	0.576
miR-20a-5p	0.948 ± 0.291	0.017*	0.002*	0.917 ± 0.265	0.083	0.963 ± 0.305	0.095
miR-22-5p	0.945 ± 0.315	0.012*	0.010*	0.891 ± 0.311	0.034*	0.971 ± 0.318	0.123
miR-30e-3p	0.973 ± 0.290	0.075	0.059	1.010 ± 0.330	0.568	0.954 ± 0.271	0.072
miR-101-3p	1.424 ± 1.241	0.256	0.265	1.744 ± 1.537	0.280	1.268 ± 1.054	0.621
miR-146a-5p	0.967 ± 0.297	0.066	0.258	0.905 ± 0.290	0.114	0.998 ± 0.298	0.345
miR-378a-3p	0.985 ± 0.248	0.172	0.397	0.974 ± 0.213	0.315	0.991 ± 0.266	0.313
miR-505-3p	0.948 ± 0.177	0.006*	0.016*	0.923 ± 0.127	0.011*	0.961 ± 0.197	0.075
Line-1 methylation	−0.306 ± 4.166	0.568	0.851	−0.448 ± 3.818	0.606	−0.238 ± 4.369	0.729

Abbreviations: SD, standard deviation, *Shows significant p-values.

Figure 2. Fitness/Ω score of the intervention group (n, 61) and the control group (n, 20) before [T0] and after [T1]. *Shows significant p-values (paired t-test).

Table 2. Distribution of the SNPs in the FTO (rs1121980), ACE (rs4341), and ACTN3 (rs1815739) gene. Genotyping values are given in total numbers and as a percentage.

Gene	SNP ID
FTO	rs1121980	Homozygotes (GG)	Homozygotes (AA)	Heterozygotes (AG)
Intervention	Total, % (n)	29.5 % (18)	24.6 % (15)	45.9 % (28)
	Male, % (n)	20.0 % (4)	30.0 % (6)	50.0 % (10)
	Female, % (n)	34.1 % (14)	22.0 % (9)	43.9 % (18)
Control	Total, % (n)	30.0 % (6)	25.0 % (5)	45.0 % (9)
	Male, % (n)	57.1 % (4)	0 % (0)	42.9 % (3)
	Female, % (n)	15.4 % (2)	38.5 % (5)	46.2 % (6)
ACE	rs4341	Homozygotes (CC)	Homozygotes (GG)	Heterozygotes (CG)
Intervention	Total, % (n)	19.7 % (12)	31.1 % (19)	49.2 % (30)
	Male, % (n)	20.0 % (4)	30.0 % (6)	50.0 % (10)
	Female, % (n)	19.5 % (8)	31.7 % (13)	48.8 % (20)
Control	Total, % (n)	5.0 % (1)	25.0 % (5)	70.0 % (14)
	Male, % (n)	14.3 % (1)	14.3 % (1)	71.4 % (5)
	Female, % (n)	0 % (0)	30.8 % (4)	69.2 % (9)
ACTN3	rs1815739	Homozygotes (CC)	Homozygotes (TT)	Heterozygotes (TC)
Intervention	Total, % (n)	32.8 % (20)	18.0 % (11)	49.2 % (30)
	Male, % (n)	20.0 % (4)	30.0 % (6)	50.0 % (10)
	Female, % (n)	39.0 % (16)	12.2 % (5)	48.8 % (20)
Control	Total, % (n)	35.0 % (7)	30.0 % (6)	35.0 % (7)
	Male, % (n)	57.1 % (4)	14.3 % (1)	28.6 % (2)
	Female, % (n)	23.1 % (3)	38.5% (5)	38.5 % (5)

Abbreviations: FTO, Fat mass and obesity-associated protein, ACE, Angiotensin-converting enzyme, ACTN3, alpha-Actinin 3.

Figure 3. Fold changes of the selected miRNAs by genotype. A) Fold change of miR-101-3p by ACE genotype (GG strength, CC endurance). B) Fold change of miR-20a-5p by ACTN3 genotype (CC strength, TT endurance). C) Fold change of miR-22-5p by ACTN3 genotype (CC strength, TT endurance). D) Fold change of miR-101-3p by ACTN3 genotype (CC strength, TT endurance). E) Fold change of miR-22-5p by FTO genotype (AA risk variant). The p-values given here are the results of the post hoc tests (Scheffé). * Shows significant p-values.

Table 3. Body composition changes through the 12-week sports intervention. Anthropometric measurements were given in mean ± SD.

	Total (n, 61)		Male (n, 20)		Female (n, 41)
	Change ± SD	p-Value	Change ± SD	p-Value	Change ± SD	p-Value
Basal metabolic rate [kcal]	23.10 ± 41.74	0.000*	20.45 ± 25.67	0.002*	24.39 ± 47.91	0.002*
Phase angel [°]	0.16 ± 0.42	0.004*	0.07 ± 0.25	0.232	0.21 ± 0.48	0.009*
LBM [kg]	0.68 ± 1.39	0.000*	0.84 ± 1.32	0.010*	0.60 ± 1.43	0.010*
BFM [kg]	−1.14 ± 1.87	0.000*	−1.33 ± 2.19	0.014*	−1.04 ± 1.72	0.000*
BCM [kg]	0.77 ± 2.16	0.007*	0.64 ± 3.17	0.378	0.83 ± 1.48	0.001*
ICW [l]	0.31 ± 0.46	0.000*	0.33 ± 0.43	0.003*	0.30 ± 0.48	0.000*
ECW [l]	0.20 ± 0.63	0.015*	0.29 ± 0.64	0.056	0.16 ± 0.63	0.112

Abbreviations: LBM, lean body mass, BFM, body fat mass, BCM, body cell mass, ICW, intracellular water, ECW, extracellular water. Fold Change, mean difference between the 2 timepoints, SD, standard deviation. *Shows significant p-values (paired t-test).

Figure 4. Significant correlations between the change of the anthropometric data and the expression of selected miRNAs, the fitness/Ω score, and Line-1 methylation after the Intervention [T1]. A) Correlation between the change of LBM and the expression of miR-22-5p at T1. B) Correlation between the change of BFM and the expression of miR-505-3p at T1. C) Correlation between the change of LBM and the expression of miR-101-3p at T1. D) Correlation between the change of BFM and the fitness/Ω score at T1. E) Correlation between the change of ECW and the Line-1 methylation at T1. Statistical significance was defined as a p-value below 0.05.

Figure 5. The properties of the measured miRNAs and their importance and classification as sports-relevant biomarkers.