Ischemia-Reperfusion Injury in a Simulated Lung Transplant Setting Differentially Regulates Transcriptomic Profiles between Human Lung Endothelial and Epithelial Cells.

Sahar Soltanieh,Yun-Bi Lu,Mingyao Liu,Junichi Sugihara,Aaron Wong,Wenyong Zhou,Ricardo Zamel,Cristina Baciu,Gaowa Saren

doi:10.3390/cells10102713

Sahar Soltanieh, Yun-Bi Lu + Show 7 more

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https://doi.org/10.3390/cells10102713

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Abstract

Current understanding of mechanisms of ischemia-reperfusion-induced lung injury during lung preservation and transplantation is mainly based on clinical observations and animal studies. Herein, we used cell and systems biology approaches to explore these mechanisms at transcriptomics levels, especially by focusing on the differences between human lung endothelial and epithelial cells, which are crucial for maintaining essential lung structure and function. Human pulmonary microvascular endothelial cells and human lung epithelial cells were cultured to confluent, subjected to different cold ischemic times (CIT) to mimic static cold storage with preservation solution, and then subjected to warm reperfusion with a serum containing culture medium to simulate lung transplantation. Cell morphology, viability, and transcriptomic profiles were studied. Ischemia-reperfusion injury induced a CIT time-dependent cell death, which was associated with dramatic changes in gene expression. Under normal control conditions, endothelial cells showed gene clusters enriched in the vascular process and inflammation, while epithelial cells showed gene clusters enriched in protein biosynthesis and metabolism. CIT 6 h alone or after reperfusion had little effect on these phenotypic characteristics. After CIT 18 h, protein-biosynthesis-related gene clusters disappeared in epithelial cells; after reperfusion, metabolism-related gene clusters in epithelial cells and multiple gene clusters in the endothelial cells also disappeared. Human pulmonary endothelial and epithelial cells have distinct phenotypic transcriptomic signatures. Severe cellular injury reduces these gene expression signatures in a cell-type-dependent manner. Therapeutics that preserve these transcriptomic signatures may represent new treatment to prevent acute lung injury during lung transplantation.

Highlights

XB130, an adaptor protein that is involved in the regulation of cell proliferation and motility in BEAS-2B cells [23,24,25], was only observed in lung epithelial cells (Figure 1E), whereas vWF and PECAM1 were not detected from epithelial cells (Figure 1D–F)
Gas exchange is the primary function of the lung; injury of alveolar epithelial and endothelial cells during IR contributes to the development of primary graft dysfunction in LTx [27]
We further developed the cell culture model that simulates major features of cold preservation and warm reperfusion with human pulmonary microvascular endothelial cells

Summary

Introduction

Lung transplantation (LTx) is a therapeutic option for patients with end-stage lung disease. Primary graft dysfunction is an important complication, contributed by ischemia-reperfusion injury (IRI). Understanding the cellular and molecular mechanisms of IRI in LTx is crucial. Multiple genes related to acute inflammation and cell death are enriched in human lung grafts during reperfusion compared with hypothermic preserved donor lungs [1]

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