Inflammatory responses relate to distinct bronchoalveolar lavage lipidome in community-acquired pneumonia patients: a pilot study

Yali Zheng, Pu Ning, Qiongzhen Luo, Yukun He, Xu Yu, Xiaohui Liu, Yusheng Chen, Xiaorong Wang, Yu Kang, Zhancheng Gao, Yali Zheng, Pu Ning, Qiongzhen Luo, Yukun He, Xu Yu, Xiaohui Liu, Yusheng Chen, Xiaorong Wang, Yu Kang, Zhancheng Gao

Abstract

Background: Community-acquired pneumonia (CAP) is a leading cause of morbidity and mortality worldwide. Antibiotics are losing their effectiveness due to the emerging infectious diseases, the scarcity of novel antibiotics, and the contributions of antibiotic misuse and overuse to resistance. Characterization of the lipidomic response to pneumonia and exploring the "lipidomic phenotype" can provide new insight into the underlying mechanisms of pathogenesis and potential avenues for diagnostic and therapeutic treatments.

Methods: Lipid profiles of bronchoalveolar lavage fluid (BALF) samples were generated through untargeted lipidomic profiling analysis using high-performance liquid chromatography with mass spectrometry (HPLC-MS). Principal component analysis (PCA) was applied to identify possible sources of variations among samples. Partitioning clustering analysis (k-means) was employed to evaluate the existence of distinct lipidomic clusters.

Results: PCA showed that BALF lipidomes differed significantly between CAP (n = 52) and controls (n = 68, including 35 healthy volunteers and 33 patients with non-infectious lung diseases); while no clear separation was found between severe CAP and non-severe CAP cases. Lactosylceramides were the most prominently elevated lipid constituent in CAP. Clustering analysis revealed three separate lipid profiles; subjects in each cluster exhibited significant differences in disease severity, incidence of hypoxemia, percentages of phagocytes in BALF, and serum concentrations of albumin and total cholesterol (all p < 0.05). In addition, SM (d34:1) was negatively related to macrophage (adjusted r = - 0.462, p < 0.0001) and PE (18:1p/20:4) was positively correlated with polymorphonuclear neutrophil (PMN) percentages of BALF (adjusted r = 0.541, p < 0.0001). The 30-day mortality did not differ amongst three clusters (p < 0.05).

Conclusions: Our data suggest that specific lower airway lipid composition is related to different intensities of host inflammatory responses, and may contribute to functionally relevant shifts in disease pathogenesis in CAP individuals. These findings argue for the need to tailor therapy based on specific lipid profiles and related inflammatory status.

Trial registration: ClinicalTrials.gov (NCT03093220). Registered on 28 March 2017 (retrospectively registered).

Keywords: Bioactive lipid; Bronchoalveolar lavage; Community-acquired pneumonia; Inflammatory response; Lipidomic profile; Phagocyte.

Conflict of interest statement

Ethics approval and consent to participate

This research was approved by the Ethical Committee of PKUPH and was conducted according to the principles expressed in the Declaration of Helsinki. All subjects provided informed consent prior to the collection of any data.

Consent for publication

All the authors have read and approved the final manuscript and the manu-script is submitted solely to Respiratory Research.

Competing interests

The authors declare that they have no competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Flowchart of study enrollment
Fig. 2
Fig. 2
Overview of bronchoalveolar lavage fluid (BALF) lipidome. a A total of 150 lipid species are detected in BALF samples of community-acquired pneumonia (CAP) and Controls. The lipids are categorized into 5 lipid classes: fatty acids (light blue), acylcarnitines (dark blue), sphingolipids (orange), neutral lipids (pink), and phospholipids (purple). b The compositions of total BALF lipid signal. Proportions of each lipid subclass are calculated by normalizing to total lipid intensities
Fig. 3
Fig. 3
Lipid profiles of CAP patients and controls. a Principal component analysis (PCA) scores plot of lipidomic profiles in BALF samples. PCA scores plot colored according to sample group: red circles, severe CAP (SCAP); green circles, non-severe CAP (NSCAP); turquoise four-point stars, healthy control (HC); and yellow triangles, quality control (QC) samples. The PCA model (R2X = 0.871, Q2 = 0.76) reflect good separation trends among SCAP and controls. Classifications based on disease severity (SCAP vs NSCAP, b), age (adult CAP vs elder CAP, c), gender (male CAP vs female CAP, d), and causative pathogens (viral, bacterial, fungal, or mixed infection, e) revealed indistinct separation trends, suggesting that the major clinical-demographic features are not the sole defining features of these BALF lipids
Fig. 4
Fig. 4
Panel a: PCA scores plot of three distinct lipid clusters (LCluster). Red, LClus1; blue, LClus2; green, LClus3. Panel b: Distinct compositions of lipid subclasses amongst three clusters. Panel c: The distributions of CAP patients with different PSI classes amongst three clusters. PSI 2 (blue section), 51–70 points; PSI 3 (green section), 71–90 points; PSI 4 (yellow section), 91–130 points; and PSI 5 (red section), 131–395 points. Panel d-e: Comparisons of macrophage percentages (d) and PMN percentages of BALF (e) amongst the three clusters. LClus1 exhibits the highest percentage of polymorphonuclear leukocytes (PMNs) and the lowest percentage of macrophages in BALF
Fig. 5
Fig. 5
Two lipid species show significantly correlations with BALF cellular components. a SM (d34:1) is inversely correlated to macrophage percentages of BALF, adjusted r = − 0.462, p < 0.0001. b PE (18:1p/20:4) is positively correlated with PMN percentages of BALF, spearman rank r = 0.3639, p < 0.0001. Red line, the fitted regression line. Areas within the grey lines, the 95% confidence intervals

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