Towards precision medicine: defining and characterizing adipose tissue dysfunction to identify early immunometabolic risk in symptom-free adults from the GEMM family study

Figures & data

Table 1. Comparison of demographic and biochemical characteristics between the n = 80 vs. n = 14 GEMM symptom-free female individuals, showing strong anthropometric and metabolic similarities and non-significant differences

Females	n = 80	n = 14
Demographic characteristics and metabolic parameters	Mean ± SD	Mean ± SD	P
Age	38.8 ± 13.34	36.39 ± 12.11	0.499
Weight (kg)	70.48 ± 15.92	67.64 ± 13.31	0.361
Waist Circunference (cm)	88.43 ± 14.24	87.46 ± 13.06	0.819
BMI (kg/m^2)	28.17 ± 5.84	28.95 ± 5.78	0.642
% Fat Total	38.58 ± 10.39	43.9 ± 5.43	0.053
Systolic Pressure (mmHg)	107.16 ± 10.48	108.14 ± 10.48	0.848
Diastolic Pressure (mmHg)	68.88 ± 8.14	69.0 ± 6.64	0.885
Glucose (mg/dL)	93.49 ± 22.92	86.71 ± 7.18	0.125
Triglycerides (mg/dL)	117.73 ± 56.77	128.78 ± 56.77	0.258
Total cholesterol (md/dL)	164.43 ± 43.09	165.07 ± 39.30	0.703
HDL- cholesterol (md/dL)	47.08 ± 14.75	43.71 ± 9.07	0.612
LDL- cholesterol (md/dL)	94.51 ± 34.22	98.71 ± 32.08	0.605

Figure 1. Mean ALR in our symptom-free female participants

Figure 2. Mean leptin, adiponectin, Adiponectin/Leptin ratio and anthropometric phenotypes of participants in the GEMM study (ALR and Age with a P-value ≤ 0.05)

Table 2. High (H: >1.8) and Low (L: <1.8) Adiponectin/Leptin ratio (ALR) according to adipokine concentrations in symptom-free female volunteers who underwent deep phenotyping. P-values for adiponectin mean levels: 0.08; leptin mean levels: 0.08; ARL: 0.001. (A p-value ≤ 0.05 is statistically significant)

Symptom-free female adults (N = 14)	Fasting Adiponectin Levels (ug/ml)	ADPN/LEP RATIO	Fasting Leptin Levels (ng/ml)
MTY0017	66.5	4.89	13.6
MTY0003	18.9	4.36	4.3
MTY0007	10.0	2.54	3.9
MTY0006	27.4	2.24	12.2
MTY0014	26.9	1.95	13.8
MEAN (H) ALR	29.9	3.2	9.6
MTY0013	13.2	1.71	7.7
MTY0020	24.1	1.48	16.3
MTY0018	23.4	1.48	15.8
MTY0019	17.2	1.40	12.2
MTY0021	21.3	1.05	20.3
MTY0009	7.5	0.90	8.4
MTY0015	15.1	0.60	25.0
MTY0016	10.0	0.36	27.5
MTY0010	3.9	0.12	32.4
MEAN (L) ALR	15.1	1.0	18.4

Figure 3. Immunometabolic and proinflammatory profile of participants in the GEMM study (non-significant P-values)

Figure 4. Immunometabolic, insulin resistance, liver enzyme profile, postprandial insulin-glucose axis and triglyceride curves of female participants in the GEMM study (n = 14). Adipo-IR Index, HOMA-IR, ALT/GPT and AST/GOT with a P-value ≤ 0.05. Area under the curve (AUC) for glucose in the (h) ALR group and (l) ALG (13,743 and 13,144, 5% difference [diff.]). AUC for insulin and GLP-1 in the (H) ALR group (5055 and 15,143) compared to the (L) ALG (9362 and 19,327) respectively, showed an 85% and 28% diff

Table 3. Multi-dimensional mass spectrometry-based shotgun lipidomics. Fed and fasted shotgun lipidomics data from specific classes considered proinflammatory: Lysophosphatidylethanolamine (LPE), Lysophosphatidylcholine (LPC) and Ceramide (Cer)

	Ceramide(Cer) (nmol/ml plasma)
Table 3	TIME	A/L R	CERN16:0	CERN18:0	CERN20:0	CERN22:0	CERN23:0	CERN24:2	CERN24:1	CERN24:0	CEROH_N24:1	CEROH_N24:0
	0 MIN	HIGH	0.34 ± 0.17	0.17 ± 0.07	0.16 ± 0.05	1.00 ± 0.22	0.99 ± 0.28	0.11 ± 0.02	1.57 ± 0.21	3.30 ± 0.95	0.23 ± 0.07	0.09 ± 0.06
		LOW	0.26 ± 0.06	0.15 ± 0.06	0.15 ± 0.05	1.01 ± 0.37	0.91 ± 0.30	0.11 ± 0.02	1.32 ± 0.37	2.89 ± 0.72	0.18 ± 0.05	0.04 ± 0.02
	30 MIN	HIGH	0.22 ± 0.05	0.15 ± 0.06	0.11 ± 0.05	0.76 ± 0.31	0.83 ± 0.32	0.09 ± 0.03	1.29 ± 0.37	2.91 ± 1.14	0.18 ± 0.08	0.05 ± 0.02
		LOW	0.21 ± 0.05	0.13 ± 0.07	0.10 ± 0.06	0.73 ± 0.39	0.72 ± 0.34	0.08 ± 0.03	1.02 ± 0.39	2.31 ± 0.95	0.14 ± 0.05	0.04 ± 0.02
	180 MIN	HIGH	0.19 ± 0.05	0.10 ± 0.06	0.06 ± 0.04	0.61 ± 0.35	0.71 ± 0.37	0.08 ± 0.05	1.04 ± 0.44	2.32 ± 1.18	0.14 ± 0.07	0.03 ± 0.02
		LOW	0.22 ± 0.08	0.13 ± 0.07	0.09 ± 0.04	0.71 ± 0.37	0.76 ± 0.33	0.09 ± 0.05	1.06 ± 0.45	2.47 ± 0.88	0.15 ± 0.04	0.04 ± 0.02
	300 MIN	HIGH	0.22 ± 0.07	0.12 ± 0.04	0.07 ± 0.02	0.60 ± 0.14	0.63 ± 0.16	0.08 ± 0.04	1.13 ± 0.39	2.54 ± 0.65	0.14 ± 0.06	0.06 ± 0.01
		LOW	0.20 ± 0.06	0.12 ± 0.07	0.10 ± 0.07	0.70 ± 0.31	0.73 ± 0.29	0.09 ± 0.04	1.02 ± 0.40	2.43 ± 0.83	0.15 ± 0.04	0.05 ± 0.04
Lysophosphatidylethanolamine (LPE) (nmol/ml plasma)
TIME	A/L R	LPEP16:1	LPEP16:0	LPE16:0	LPEP18:1	LPEP18:0	LPE18:3	LPE18:2	LPE18:1	LPE18:0	LPE20:4	LPE20:3	LPE22:6
0 MIN	HIGH	0.69 ± 0.44	4.87 ± 2.30	9.75 ± 2.73	2.47 ± 0.96	4.47 ± 3.01	0.30 ± 0.10	7.72 ± 2.88	5.60 ± 1.65	11.51 ± 3.22	9.77 ± 5.47	1.00 ± 0.44	2.35 ± 1.15
	LOW	0.57 ± 0.43	3.80 ± 1.27	7.76 ± 2.31	2.27 ± 0.94	3.95 ± 1.33	0.36 ± 0.13	8.12 ± 1.93	5.53 ± 1.53	9.59 ± 1.88	11.52 ± 5.37	1.21 ± 0.40	2.43 ± 1.13
30 MIN	HIGH	0.21 ± 0.12	1.95 ± 0.12	8.85 ± 3.38	0.95 ± 0.46	2.01 ± 1.01	0.30 ± 0.10	7.93 ± 2.56	5.25 ± 1.72	10.44 ± 4.57	7.03 ± 1.74	0.91 ± 0.19	2.16 ± 0.86
	LOW	0.28 ± 0.31	2.04 ± 1.51	6.43 ± 1.87	1.10 ± 0.97	1.88 ± 1.40	0.30 ± 0.14	8.15 ± 2.86	4.89 ± 1.51	7.35 ± 1.65	8.21 ± 4.10	0.97 ± 0.51	1.97 ± 0.79
180 MIN	HIGH	0.18 ± 0.11	2.20 ± 0.69	8.18 ± 1.76	1.06 ± 0.30	2.09 ± 0.41	0.34 ± 0.21	7.15 ± 1.05	5.36 ± 0.63	9.36 ± 2.01	5.02 ± 2.88	0.63 ± 0.26	1.06 ± 0.60
	LOW	0.28 ± 0.90	2.71 ± 1.73	8.56 ± 2.80	1.28 ± 0.98	2.47 ± 1.45	0.43 ± 0.11	10.34 ± 2.64	7.37 ± 1.62	10.07 ± 3.23	6.32 ± 2.88	0.94 ± 0.47	1.38 ± 0.70
300 MIN	HIGH	0.27 ± 0.10	2.30 ± 0.37	10.00 ± 4.65	1.00 ± 0.11	2.06 ± 0.25	0.63 ± 0.39	10.12 ± 4.11	7.08 ± 2.44	10.46 ± 2.90	7.29 ± 4.30	0.95 ± 0.46	1.87 ± 1.36
	LOW	0.22 ± 0.06	3.86 ± 3.90	8.73 ± 6.24	1.63 ± 1.45	3.56 ± 4.03	0.47 ± 0.17	9.54 ± 2.45	7.47 ± 2.00	12.25 ± 10.09	6.36 ± 3.82	0.95 ± 0.55	1.45 ± 0.81
Lyso Phosphatidylcholine (LPC) (nmol/ml plasma)
TIME	A/L R	LPCP16:0	LPCA16:0	LPC16:1	LPC16:0	LPC18:3	LPC18:2	LPC18:1	LPC18:0	LPC20:4	LPC20:3	LPC20:2	LPC20:1
0 MIN	HIGH	37.12 ± 38.87	3.14 ± 0.82	21.54 ± 15.27	120.54 ± 31. 07	0.76 ± 0.18	42.41 ± 12.88	24.43 ± 5.83	27.07 ± 7.00	9.28 ± 2.61	2.99 ± 0.63	0.93 ± 0.39	0.52 ± 0.15
	LOW	22.23 ± 15.58	2.50 ± 0.38	18.24 ± 9.30	96.82 ± 11.93	0.86 ± 0.36	39.87 ± 10.54	20.40 ± 5.39	21.63 ± 3.43	7.14 ± 1.91	2.69 ± 0.69	1.08 ± 0.40	0.70 ± 0.79
30 MIN	HIGH	4.23 ± 5.18	2.55 ± 0.66	6.20 ± 3.38	114.05 ± 32.40	0.83 ± 0.22	45.16 ± 15.94	23.90 ± 6.06	25.01 ± 7.90	9.46 ± 3.26	3.07 ± 0.92	1.32 ± 0.33	0.58 ± 0.09
	LOW	8.36 ± 21.27	2.12 ± 0.51	8.55 ± 13.11	87.32 ± 12.58	0.78 ± 0.30	39.14 ± 10.90	18.94 ± 5.82	18.70 ± 3.15	6.64 ± 1.67	2.67 ± 1.05	1.29 ± 0.61	0.47 ± 0.13
180 MIN	HIGH	1.35 ± 0.94	2.58 ± 0.70	5.21 ± 2.16	117.18 ± 27.66	0.90 ± 0.28	48.06 ± 13.71	22.34 ± 5.15	23.34 ± 4.99	9.24 ± 3.33	2.94 ± 1.23	0.98 ± 0.78	0.53 ± 0.35
	LOW	2.59 ± 3.34	2.42 ± 0.63	5.53 ± 2.73	107.69 ± 26.04	0.99 ± 0.43	53.03 ± 16.75	22.87 ± 8.54	22.36 ± 5.25	7.56 ± 1.79	2.99 ± 1.36	1.23 ± 0.57	0.52 ± 0.17
300 MIN	HIGH	2.49 ± 3.54	2.34 ± 0.76	4.93 ± 2.87	106.19 ± 38.14	0.91 ± 0.33	52.80 ± 22.19	23.03 ± 8.09	22.13 ± 7.34	9.50 ± 3.72	2.99 ± 1.12	0.95 ± 0.35	0.53 ± 0.19
	LOW	2.49 ± 3.12	2.46 ± 1.39	5.80 ± 3.59	113.44 ± 71.40	0.97 ± 0.47	52.19 ± 16.15	23.00 ± 9.46	24.93 ± 17.73	8.29 ± 2.75	3.18 ± 1.71	0.93 ± 0.60	0.43 ± 0.20

Table 4. Plasma MicroRNA (miR) in symptom-free females with (H) and (L) ALR. Fed and fasted circulating miRNAs selected from recent literature with adipogenesis-promoting or anti-adipogenic function. miR-27b-5p (fasting and postprandial), miR-375 (postprandial), miR-140-5p (postprandial), miR-130b-3p (fasting and postprandial) with a P-value ≤ 0.05

		Adiponectin/leptin ratio
Table 4		TIME 0 (Fasting)			TIME 180 (Postprandial)
FUNCTION	miRNA	HIGH	LOW	P	HIGH	LOW	P
ADIPOGENESIS PROMOTING miRNA	miR-27b-5p	41.2 ± 26.7	65.1 ± 14.6	0.02	41.3 ± 26.6	64.3 ± 16.0	0.04
	miR-378a-3p	54.5 ± 39.9	86.7 ± 15.5	0.11	52.7 ± 37.8	86.1 ± 11.1	0.11
	miR-375	18.1 ± 12.2	35.0 ± 16.9	0.08	17.9 ± 11.3	41.7 ± 21.3	0.01
	miR-140-5p	/	/	/	11.0 ± 6.4	16.7 ± 2.1	0.02
ANTI-ADIPOGENIC miRNA	miR-33a-5p	17.5 ± 8.3	28.4 ± 15.0	0.23	18.8 ± 6.6	27.4 ± 15.0	0.11
	miR-130b-3p	24.8 ± 16.3	41.2 ± 8.0	0.01	23.6 ± 15.7	48.1 ± 8.5	0.02
	miR-7-1-3p	0.006 ± 0.01	0.4 ± 0.5	0.06	0.005 ± 0.01	0.4 ± 0.5	0.06
	let-7a-3p	/	/	/	20.9 ± 14.3	35.0 ± 15.3	0.08

Figure 5. Direct measurements performed for fasting (0ʹ) and fed (180ʹ) subcutaneous fat transcriptomic profiling looking for molecular trends in adipose tissue dysfunction or inflammation differential gene expression in key proinflammatory and anti-inflammatory genes. The expression of the anti-inflammatory gene ANG was significantly decreased in the (l)ALR subgroup in fasting