Elevated Fibronectin Levels in Profibrotic CD14+ Monocytes and CD14+ Macrophages in Systemic Sclerosis.

Michał Rudnik,Oliver Distler,Janine Schniering,Ievgeniia Kocherova,Carol Feghali-Bostwick,Przemysław Błyszczuk,Martin Ehrbar,Gabriela Kania,Karin Klingel,Amela Hukara,Vincent Milleret,Suzana Jordan

doi:10.3389/fimmu.2021.642891

Abstract

BackgroundSystemic sclerosis (SSc) is an autoimmune disease characterized by overproduction of extracellular matrix (ECM) and multiorgan fibrosis. Animal studies pointed to bone marrow-derived cells as a potential source of pathological ECM-producing cells in immunofibrotic disorders. So far, involvement of monocytes and macrophages in the fibrogenesis of SSc remains poorly understood.Methods and ResultsImmunohistochemistry analysis showed accumulation of CD14+ monocytes in the collagen-rich areas, as well as increased amount of alpha smooth muscle actin (αSMA)-positive fibroblasts, CD68+ and mannose-R+ macrophages in the heart and lungs of SSc patients. The full genome transcriptomics analyses of CD14+ blood monocytes revealed dysregulation in cytoskeleton rearrangement, ECM remodeling, including elevated FN1 (gene encoding fibronectin) expression and TGF-β signalling pathway in SSc patients. In addition, single cell RNA sequencing analysis of tissue-resident CD14+ pulmonary macrophages demonstrated activated profibrotic signature with the elevated FN1 expression in SSc patients with interstitial lung disease. Peripheral blood CD14+ monocytes obtained from either healthy subjects or SSc patients exposed to profibrotic treatment with profibrotic cytokines TGF-β, IL-4, IL-10, and IL-13 increased production of type I collagen, fibronectin, and αSMA. In addition, CD14+ monocytes co-cultured with dermal fibroblasts obtained from SSc patients or healthy individuals acquired a spindle shape and further enhanced production of profibrotic markers. Pharmacological blockade of the TGF-β signalling pathway with SD208 (TGF-β receptor type I inhibitor), SIS3 (Smad3 inhibitor) or (5Z)-7-oxozeaenol (TGF-β-activated kinase 1 inhibitor) ameliorated fibronectin levels and type I collagen secretion.ConclusionsOur findings identified activated profibrotic signature with elevated production of profibrotic fibronectin in CD14+ monocytes and CD14+ pulmonary macrophages in SSc and highlighted the capability of CD14+ monocytes to acquire a profibrotic phenotype. Taking together, tissue-infiltrating CD14+ monocytes/macrophages can be considered as ECM producers in SSc pathogenesis.

Highlights

Systemic sclerosis (SSc) is an autoimmune disease, characterized by high morbidity and mortality and a significant reduction in quality of life
We evaluated the differentiation potential of circulating CD14+ monocytes from healthy controls and SSc patients into a myofibroblast-like phenotype
After integration we reduced the dimensionality of the data using Principal Component Analysis (PCA)

Summary

Introduction

Systemic sclerosis (SSc) is an autoimmune disease, characterized by high morbidity and mortality and a significant reduction in quality of life. Microvascular damages, dysregulation of innate and adaptive immunity and multiorgan fibrosis are implicated in the pathophysiology of SSc [1]. SSc is characterized by a patientto-patient variability in clinical manifestations, autoantibody profiles, extent of disease, treatment response and reduced survival rate [2]. Fibrogenesis is a multistage process, considered as the result of impaired tissue repair responses, in which abnormal production of cytokines, growth factors and angiogenic factors turn tissue homeostasis towards the excessive accumulation of extracellular matrix (ECM) [4]. Systemic sclerosis (SSc) is an autoimmune disease characterized by overproduction of extracellular matrix (ECM) and multiorgan fibrosis. Involvement of monocytes and macrophages in the fibrogenesis of SSc remains poorly understood

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