Nasal Lavage Fluid Proteomics Reveals Potential Biomarkers of Asthma Associated with Disease Control

Figures & data

Table 1 Demographic Data of Study Participants

Characteristic	HCs	Patients with Allergic Disease				P-HCs vs Patients	P-AR vs Asthma vs AR+Asthma
		Total	AR	Asthma	AR+Asthma
Subjects (no.)	12	88	25	24	27
Age (y), mean ± SEM	37.58±11.22	40.50±13.42	39.08±11.07	43.04±14.97	39.56±14.11	0.477	0.535
Sex ratio (M/F)	8/4	36/40	9/16	15/9	12/15	0.214	0.166
Smoking status						0.186	0.664
Current smoker (%)	3 (25.00)	9 (10.23)	1 (4.00)	4 (16.67)	4 (14.80)
Quitting	0	13 (14.77)	4 (16.00)	4 (16.67)	5 (18.50)
Never smoke	9 (75.00)	54 (61.36)	20 (80.00)	16 (16.67)	18 (66.70)
BMI (kg/m^2)	23.18±3.31	22.40±3.19	23.51±2.78	21.88±3.45	21.83±3.16	0.433	0.104
FEV1 (% predicted)	97.93±11.94	86.35±22.87	101.70±13.71	75.12±24.11	82.11±21.44	0.013	<0.001
FEV1/FVC (%)	84.50±4.73	78.68±13.86	86.95±6.82	73.77±15.63	75.38±14.00	0.008	0.001
PEF (%)	98.87±14.64	85.16±22.42	93.82±10.81	77.98±25.69	83.53±25.22	0.044	0.069
MMEF75/25 (%)	83.96±19.68	66.29±33.70	87.75±21.62	53.82±37.86	57.49±30.17	0.017	<0.001
ACQ	0	0.95±1.07	NA	1.35±1.00	1.49±1.07	<0.001	0.642
AQLQ	244.58±1.00	224.01±21.70	NA	223.25±12.44	205.26±20.78	<0.001	<0.001

Abbreviations: HCs, healthy controls; AR, allergic rhinitis; M, male; F, female; ACQ, the Asthma Control Questionnaire; AQLQ, the Asthma Quality of Life Questionnaire; NA, not applicable.

Table 2 The List of Proteins Upregulated in Nasal Lavage Fluid Between AR, Asthma, and Asthma +AR as Compared with HCs

Gene Names	Protein Names	Function
CD55	CD55	Cell membrane; complement pathway; disulfide bond; immunity; innate immunity; membrane; receptor
ANPEP	CD13	Angiogenesis; cell membrane; host cell receptor for virus entry; host–virus interaction; involved the cleavage of peptides bound to major histocompatibility complex class II molecules of antigen presenting cells.
ALPL	Alkaline phosphatase	Calcium; cell membrane; direct protein sequencing; disease variant; disulfide bond; glycoprotein; hydrolase; lipoprotein; magnesium; membrane
SOD2	Superoxide dismutase	Oxidoreductase; acetylation; direct protein sequencing; manganese; mitochondrion; nitration; reference proteome; transit peptide
KLK13	Kallikrein-13	Alternative splicing; disulfide bond; glycoprotein; hydrolase; protease; reference proteome; secreted; serine protease; signal
SERPINB7	Serpin B7	Alternative splicing; cytoplasm; palmoplantar keratoderma; phosphoprotein; protease inhibitor; reference proteome; serine protease inhibitor
BLMH	Bleomycin hydrolase	Cytoplasm; direct protein sequencing; hydrolase; protease; reference proteome
C3	Complement C3	Complement alternate pathway; complement pathway
C5	Complement C5	Complement alternate pathway; complement pathway
C7	Complement C7	Complement alternate pathway; complement pathway

Figure 1 Differentially expressed proteins (DEPs) among AR, asthma, combined asthma+ AR and healthy controls (HCs) for nasal lavage fluid. Volcano plot of DEPs, (A) AR vs HCs; (B) asthma vs HCs; (C) combined asthma+ AR vs HCs; (D) Venn diagram of DEPs. Fold change ≥2.0 or fold change ≤0.5 (false-discovery rate, FDR <0.05). AR, allergic rhinitis, HCs, healthy controls.

Figure 2 DEPs among AR, asthma, combined asthma +AR, and HCs (A) CD55, (B) C3, (C) C5, (D) C7, (E) ANPEP, (F) ALPL, (G) SOD2, (H) SERPINB7, (I) KLK13 AR, allergic rhinitis, HCs, healthy controls, DEPs, differentially expressed proteins. *P < 0.05, **P < 0.01, ***P < 0.001.

Figure 3 Proteomic analysis in patients with asthma according to asthma control. (A) Volcano plot of DEPs, Fold change ≥1.5 or ≤0.66 (P < 0.05), (B) Pearson correlation heat maps of DEPs, (C) STRING protein–protein interaction (PIP) network based on DEPs (P < 0.05), (D) the abundance of DEPs. DEPs, differentially expressed proteins. *P < 0.05, **P < 0.01.

Figure 4 Proteomic analysis in patients with combined asthma +AR according to asthma control. (A) Volcano plot of DEPs. Fold change ≥1.5 or ≤0.66 (P < 0.05), (B) Pearson correlation heat maps of DEPs, (C) the abundance of DEPs. DEPs, differentially expressed proteins. *P < 0.05, **P < 0.01, *** P < 0.001.

Data Sharing Statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.