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The exosomal CD44 from bronchoalveolar lavage fluid as a biomarker for pulmonary fibrosis in diffuse parenchymal lung diseases

ABSTRACT: Diffuse parenchymal lung diseases (DPLD) cover heterogeneous types of lung disorders with a plethora of variously combined clinical manifestations. Among many pathological phenotypes, pulmonary fibrosis is the most devastating. Pulmonary fibrosis is a characteristic sign for idiopathic pulmonary fibrosis (IPF); however, it may occur also in later stages of other types of DPLD. Despite a poor prognosis brought by pulmonary fibrosis, there are no specific diagnostic biomarkers for the initial development of this fatal condition. The major hallmark of lung fibrosis is uncontrolled activation of lung fibroblasts to myofibroblasts associated with extracellular matrix deposition and the loss of both lung structure and function. Here, we used this peculiar feature in order to identify specific biomarkers of pulmonary fibrosis in bronchoalveolar lavage fluids. The primary MRC-5 human fibroblasts were activated with BALF collected from patients with clinically diagnosed lung fibrosis; the activated fibroblasts were then washed rigorously, and further incubated to allow secretion; afterwards, the secretomes were analysed by mass spectrometry. In this way, the CD44 protein was identified. Finally, BALF of all DPLD patients were positively tested for the presence of CD44 by ELISA. Biochemical and biophysical characterizations revealed an exosomal origin of CD44. Correlation studies confirmed the exosomal CD44 in BALF as a specific and reliable biomarker of IPF and other types of DPLD accompanied with pulmonary fibrosis.

INSTRUMENT(S): Orbitrap Exploris 480

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Mrc-5 Cell

SUBMITTER: Peter Barath

LAB HEAD: Peter Barath

PROVIDER: PXD055250 | Pride | 2025-01-03

REPOSITORIES: Pride

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			Action	DRS
	20240219_metoda17_M1_controlPBS_20ul_meranie3.raw	Raw
	20240219_metoda17_M1_controlPBS_meranie1.raw	Raw
	20240219_metoda17_M2_controlPBS_meranie1.raw	Raw
	20240219_metoda17_M2_controlPBS_meranie2.raw	Raw
	20240219_metoda17_M3_IPF_meranie1.raw	Raw

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Project description:Objective: Pulmonary complications in systemic sclerosis (SSc), including pulmonary fibrosis (PF) and pulmonary arterial hypertension (PAH), are the leading cause of mortality. We compared the molecular fingerprint of SSc lung tissues and matching primary lung fibroblasts to those of normal donors, and patients with idiopathic pulmonary fibrosis (IPF) and idiopathic pulmonary arterial hypertension (IPAH). Methods: Lung tissues were obtained from 33 patients with SSc who underwent lung transplantation. Tissues and cells from a subgroup of SSc patients with predominantly PF or PAH were compared to those from normal donors, patients with IPF, or IPAH. Microarray data was analyzed using Efficiency Analysis for determination of optimal data processing methods. Real time PCR and immunohistochemistry were used to confirm differential levels of mRNA and protein, respectively. Results: We identified a consensus of 242 and 335 genes that were differentially expressed in lungs and primary fibroblasts, respectively. Enriched function groups in SSc-PF and IPF lungs included fibrosis, insulin-like growth factor signaling and caveolin-mediated endocytosis. Functional groups shared by SSc-PAH and IPAH lungs included antigen presentation, chemokine activity, and IL-17 signaling. Conclusion: Using microarray analysis on carefully phenotyped SSc and comparator lung tissues, we demonstrated distinct molecular profiles in tissues and fibroblasts of patients with SSc-associated lung disease compared to idiopathic forms of lung disease. Unique molecular signatures were generated that are disease- (SSc) and phenotype- (PF vs PAH) specific. These signatures provide new insights into pathogenesis and potential therapeutic targets for SSc lung disease. Lung tissues were obtained from 33 patients with SSc who underwent lung transplantation. Tissues and cells from a subgroup of SSc patients with predominantly PF or PAH were compared to those from normal donors, patients with IPF, or IPAH. Microarray data was analyzed using Efficiency Analysis for determination of optimal data processing methods. Real time PCR and immunohistochemistry were used to confirm differential levels of mRNA and protein, respectively.

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The exosomal CD44 from bronchoalveolar lavage fluid as a biomarker for pulmonary fibrosis in diffuse parenchymal lung diseases

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