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

Abstract

Purpose

Little research has explored the proteomic characteristics of nasal lavage fluid from asthmatic patients. This study aims to investigate whether differentially expressed proteins (DEPs) in nasal lavage fluid can serve as a biomarker to differentiate asthma patients from healthy controls (HCs) and to discern between individuals with well controlled and poorly controlled asthma.

Patients and Methods

We enrolled patients with allergic rhinitis (AR), asthma, or both conditions, and HCs in this study. We recorded patients’ demographic and medical history data and administered asthma quality of life questionnaire (AQLQ) and asthma control questionnaire (ACQ). Nasal fluid samples were collected, followed by protein measurements, and proteomic analysis utilizing the data-independent acquisition (DIA) method.

Results

Twenty-four with asthma, 27 with combined asthma+ AR, 25 with AR, and 12 HCs were enrolled. Four proteins, superoxide dismutase 2 (SOD2), serpin B7 (SERPINB7), kallikrein-13 (KLK13), and bleomycin hydrolase (BLMH) were significantly upregulated in nasal lavage fluid samples of asthma without AR, compared to HCs (Fold change ≥2.0, false-discovery rate [FDR] <0.05). Conversely, 56 proteins including secretoglobin family 2A member 1 (SCGB2A1) were significantly downregulated (fold change ≥2.0, FDR <0.05). Furthermore, 96.49% of DEPs including peptidase inhibitor 3 (PI3) and C-X-C motif chemokine 17 (CXCL17) were upregulated in poorly controlled asthma patients without AR relative those with well- or partly controlled asthma (fold change ≥1.5, FDR <0.05). Search tool for the retrieval of interacting genes/proteins (STRING) analysis showed that PI3, with 18 connections, may be pivotal in asthma control.

Conclusion

The study revealed significant alteration in the nasal lavage proteome in asthma without AR patients. Moreover, our results indicated a potential association between the expression of proteome in the upper airway and the level of asthma control. Specifically, PI3 appears to be a key role in the regulation of asthma without AR.

Graphical Abstract

Keywords:

• allergy
• proteomic analysis
• nasal lavage fluid
• asthma control

Abbreviations

ACQ, asthma control questionnaire; ALPL, alkaline phosphatase; AMBP, alpha1-microglobulin bikunin; ANPEP, aminopeptidase N; AQLQ, asthma quality of life questionnaire; AR, allergic rhinitis; AUC, area under curve; AZGP1, Zinc-alpha-2-glycoprotein; BLMH, bleomycin hydrolase; BMI, body mass index; C3, complement C3; CALML5, calmodulin-like protein 5; CASP14, caspase-14; CCL, C-C motif ligand; CD59, CD59 glycoprotein; CDSN, corneodesmosin; C1R, complement C1r subcomponent; CI, confidence intervals; CXCL17, C-X-C motif chemokine 17; CXCL8, C-X-C motif chemokine ligand 8; CD55 (DAF), decay-accelerating factor; DBNL, drebrin-like protein; DCD, dermcidin; DEFB1, beta-defensin 1; DEPs, differentially expressed proteins; DSC1, desmocollin-1; DC, dendritic cell; DIA, data-independent acquisition; DSC2, desmocollin-2; DSTN, destrin; ELISA, enzyme-linked immunosorbent assay; FDR, false-discovery rate; FEV1, forced expiratory volume in 1 second; FLG, filaggrin; FVC, forced vital capacity; GO, Gene Ontology; HBB, hemoglobin subunit beta; HCs, healthy controls; HDAC2, histone deacetylase 2; HP, haptoglobin; IGFALS, insulin-like growth factor-binding protein complex acid labile subunit; IGFBP2, insulin-like growth factor-binding protein 2; IGHV3-49, immunoglobulin heavy variable 3-49; IL, interleukin; iRT, indexed retention time; iTRAQ, isobaric tags for relative and absolute quantitation; KLK13, kallikrein-13; KLKB1, plasma kallikrein; KNG1, kininogen-1; KRT16, keratin, type I cytoskeletal 16; KRT6A, keratin, type II cytoskeletal 6A; KRT78, keratin, type II cytoskeletal 78; LYZ, lysozyme C; MDK, midkine; MMEF75/25, maximal mid-expiratory flow; MMP10, stromelysin-2; MUC4, mucin-4; NOS2, nitric oxide synthase, inducible; PEF, peak expiratory flow; PGM1, phosphoglucomutase-1; PGM2, phosphoglucomutase-2; PI3, peptidase inhibitor 3; PIP, prolactin-inducible protein; PLA2G2A, phospholipase A2, membrane associated; POSTIN, periostin; PPI, protein–protein interaction; PRB2, basic salivary proline-rich protein 2; PRB4, basic salivary proline-rich protein 4; PROS1, protein S; PYGL, glycogen phosphorylase, liver form; QPCT, glutaminyl-peptide cyclotransferase; RAB1A, ras-related protein Rab-1A; ROC, receiver operating characteristic; S100A12, protein S100-A12; SCGB1A1, uteroglobin; SCGB1D1, secretoglobin family 1D member 1; SCGB1D2, secretoglobin family 1D member 2; SCGB2A1, secretoglobin family 2A member 1; SEMG2, semenogelin-2; SERPINA1, alpha-1-antitrypsin; SERPINB3, serpin B3; SERPINB7, serpin B7; SERPINI1, neuroserpin; SLPI, antileukoproteinase; SOD2, superoxide dismutase 2; SPRR1B, cornifin-B; STRING, search tool for the retrieval of interacting genes/proteins; TGFBI, transforming growth factor-beta-induced protein ig-h3; Th, T-helper type; TIMP2, metalloproteinase inhibitor 2; TNSS, total nasal symptom score; TSLP, thymic stromal lymphopoietin; WFDC2, WAP four-disulfide core domain protein 2.

Data Sharing Statement

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

Ethics Approval and Informed Consent

This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the ethics committee of the First Affiliated Hospital of Ningbo University (approval 2020-R145). All participants agreed and signed informed consents before enrolment.

Author Contributions

All authors made a significant contribution to the work reported, whether that is in the conception, study design, execution, acquisition of data, analysis and interpretation, or in all these areas; took part in drafting, revising or critically reviewing the article; gave final approval of the version to be published; have agreed on the journal to which the article has been submitted; and agree to be accountable for all aspects of the work.

Disclosure

The authors report no conflicts of interest in this work.

Additional information

Funding

This research was supported by the National Natural Science Foundation of China (Grant number [82170016]).