Gut Microbiome and Metabolomics Profiles of Allergic and Non-Allergic Childhood Asthma

Figures & data

Table 1 Baseline Characteristics of All Study Populations

Variable	Allergic Asthmatics (n=27)	Non-Allergic Asthmatics (n=10)	Healthy Control (n=20)	P value
Gender				0.080
Boys, n (%)	15 (55.6)	4 (40.0)	9 (45.0)
Girls, n (%)	12 (44.4)	6 (60.0)	11 (55.0)
Age (years, mean ± SD)	6.9 ± 1.4	9.3 ± 2.8	8.9 ± 1.5	< 0.010
BMI (kg/m^2, mean ± SD)	16.5 ± 1.8	18.7 ± 2.9	17.9 ± 3.5	0.298
High-fiber food intakes^a (times/week, mean ± SD)	8.9 ± 3.5	10.6 ± 3.2	11.4 ± 5.6	0.150
High-protein food intakes^b (times/week, mean ± SD)	4.5 ± 0.6	3.8 ± 1.5	4.5 ± 0.9	0.107

Notes: ^aHigh-fiber food included vegetables, mushrooms and fruit. ^bHigh-protein food included eggs and duck eggs.

Abbreviations: BMI, body mass index; SD, standard deviation.

Table 2 Clinical Information of Asthmatic Children

Variable	Allergic Asthmatics (n=27)	Non-Allergic Asthmatics (n=10)
Lung function (%, mean±SD)^a
FEV1	96.2±16.1	85.9±21.3
FVC	97.6± 7.0	90.8±19.7
MMEF75/25	75.3± 29.0	61.2±20.8
FeNO (ppb, mean±SD)	39.1± 31.1	21.3±11.9
Allergens foy skin pick testing
Cat and dog allergies, n (%)	7 (25.9)	None
Mites, n (%)	11 (40.7)	None
Pollens of weeds, trees and grasses, n (%)	13 (48.2)	None
Fungi, n (%)	11 (40.7)	None

Note: ^aLung function was the percentage of measured value to predicted value.

Abbreviations: SD, standard deviation; FEV1, forced expiratory volume in the first second; FVC, forced vital capacity; MMEF, maximal mid-expiratory flow; FeNO, fractional exhaled nitric oxide; ppb, part per billion.

Figure 1 Continued.

Figure 1 Characters of gut microbiome. (A and B) Alpha diversity (Chao1 and Simpson parameter) in allergic, non-allergic asthma and healthy control groups. (C) PCoA plot based on weighted UniFrac distance, (D) PCoA plot based on unweighted UniFrac distance. (E) Relative abundance of top 10 phyla by groups. The height of bars represents relative abundance. (F) LEfSe analysis between allergic asthma and healthy control groups. (G) LEfSe analysis between non-allergic asthma and healthy control groups. The graph of LEfSe distinguished the microbial communities of each group with LDA > 2 and p < 0.05.

Abbreviations: AA, allergic asthma group; NA, non-allergic asthma group; HC, healthy control group.

Table 3 Genera with Significant Differences in Relative Abundance Between Asthmatic Group and Healthy Control Group (Wilcoxon Test)

Genus	Affiliated Phylum	Affiliated Class	Affiliated Order	Affiliated Family	Asthmatics	Healthy Control	P value
					Mean ± SD^a	Mean ± SD^a
Allergic asthmatic group vs Healthy control group
Prevotella	Bacteroidetes	Bacteroidia	Bacteroidales	Prevotellaceae	1.27±5.26	4.19±9.41	<0.001
Lactobacillus	Firmicutes	Bacilli	Lactobacillales	Lactobacillaceae	0.42±0.42	0.09±0.12	0.006
Lachnospiraceae ND3007 group	Firmicutes	Clostridia	Lachnospirales	Lachnospiraceae	0.53±0.50	0.79±0.28	0.040
Collinsella	Actinobacteriota	Coriobacteriia	Coriobacteriales	Coriobacteriaceae	0.54±0.88	0.90±0.62	0.011
Terrisporobacter	Firmicutes	Clostridia	Peptostreptococcales. Tissierellales	Peptostreptococcacea	0.42±0.80	1.32±1.90	0.002
Megamonas	Firmicutes	Negativicutes	Veillonellales. Selenomonadales	Selenomonadaceae	0.15±0.17	0.26±0.19	0.005
Eubacterium eligens group	Firmicutes	Clostridia	Lachnospirales	Lachnospiraceae	0.26±0.30	0.82±0.35	<0.001
CAG.56	Firmicutes	Clostridia	Lachnospirales	Lachnospiraceae	0.26±0.35	0.38±0.27	0.017
Lachnospira	Firmicutes	Clostridia	Lachnospirales	Lachnospiraceae	0.30±0.54	0.44±0.17	0.003
Non-allergic asthmatic group vs Healthy control group
Bacteroides	Bacteroidetes	Bacteroidia	Bacteroidales	Bacteroidaceae	10.08±6.28	20.35±7.09	0.003
Agathobacter	Firmicutes	Clostridia	Lachnospirales	Lachnospiraceae	1.72±1.64	3.38±2.16	0.043
CAG.352	Firmicutes	Clostridia	Oscillospirales	Ruminococcaceae	1.31±1.00	2.82±1.61	0.029
Ruminococcus	Firmicutes	Clostridia	Oscillospirales	Ruminococcaceae	0.95±0.85	1.82±0.80	0.029
Intestinibacter	Firmicutes	Clostridia	Peptostreptococcales. Tissierellales	Peptostreptococcaceae	0.81±0.74	1.14±0.33	0.029
Lachnoclostridium	Firmicutes	Clostridia	Lachnospirales	Lachnospiraceae	0.42±0.14	0.68±0.20	0.003
Lactobacillus	Firmicutes	Bacilli	Lactobacillales	Lactobacillaceae	1.86±4.00	0.09±0.12	0.012
Turicibacter	Firmicutes	Bacilli	Erysipelotrichales	Erysipelotrichaceae	0.40±0.35	1.03±1.42	0.005
Lachnospiraceae ND3007 group	Firmicutes	Clostridia	Lachnospirales	Lachnospiraceae	0.46±0.55	0.79±0.28	0.023
Collinsella	Actinobacteriota	Coriobacteriia	Coriobacteriales	Coriobacteriaceae	0.27±0.30	0.90±0.62	0.009
Terrisporobacter	Firmicutes	Clostridia	Peptostreptococcales. Tissierellales	Peptostreptococcaceae	0.12±0.10	1.32±1.90	<0.001
Parasutterella	Proteobacteria	Gammaproteo-bacteria	Burkholderiales	Sutterellaceae	0.23±0.25	0.34±0.24	0.043
Eubacterium eligens group	Firmicutes	Clostridia	Lachnospirales	Lachnospiraceae	0.18±0.14	0.82±0.35	<0.001
Lachnospira	Firmicutes	Clostridia	Lachnospirales	Lachnospiraceae	0.11±0.09	0.44±0.17	<0.001

Note: ^aRelative abundance, %.

Abbreviation: SD, standard deviation.

Table 4 The Involved Metabolic Pathways of the Altered Metabolites in Allergic and Non-Allergic Asthma Groups

Pathway	Allergic Asthmatics (Up, Down)	Non-Allergic Asthmatics (Up, Down)
Amino Acid	8 (1,7)	15 (4,11)
Lipid	10 (5,5)	19 (1,18)
Peptide	7 (0,7)	2 (0,2)
Carbohydrate	0	1 (0,1)
Energy	2 (2,0)	0
Nucleotide	5 (1,4)	4 (0,4)
Cofactors and Vitamins	2 (1,1)	4 (2,2)
Xenobiotics	8 (4,4)	13 (2,11)

Figure 2 Continued.

Figure 2 Continued.

Figure 2 Continued.

Figure 2 Continued.

Figure 2 Spearman correlation heatmap for significantly altered metabolites and genera in asthmatics. *p <0.05, **p <0.01. (A) Correlation heatmap for amino acid. (B) Correlation heatmap for Lipid. (C) Correlation heatmap for peptide. (D) Correlation heatmap for nucleotide. (E) Correlation heatmap for others, including carbohydrate, energy, cofactors and vitamins.