Acellular ex vivo lung perfusate silences pro-inflammatory signaling in human lung endothelial and epithelial cells

Jamie E Jeon; Lei Huang; Zhiyuan Zhu; Aaron Wong; Shaf Keshavjee; Mingyao Liu

doi:10.1186/s12967-023-04601-w

Acellular ex vivo lung perfusate silences pro-inflammatory signaling in human lung endothelial and epithelial cells

J Transl Med. 2023 Oct 17;21(1):729. doi: 10.1186/s12967-023-04601-w.

Authors

Jamie E Jeon^#^{1

2}, Lei Huang^#^{1

3}, Zhiyuan Zhu^#^{1

4}, Aaron Wong^{1

5}, Shaf Keshavjee^{1

5

6}, Mingyao Liu^{7

8

9}

Affiliations

¹ Latner Thoracic Surgery Research Laboratories, Toronto General Hospital Research Institute, University Health Network, 101 College Street, PMCRT2-814, Toronto, ON, M5G 1L7, Canada.
² Department of Physiology, Temerty Faculty of Medicine, University of Toronto, Toronto, ON, Canada.
³ Department of Thoracic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
⁴ Department of Otolaryngology, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, 210000, China.
⁵ Institute of Medical Science, Temerty Faculty of Medicine, University of Toronto, Toronto, ON, Canada.
⁶ Department of Surgery, Temerty Faculty of Medicine, University of Toronto, Toronto, ON, Canada.
⁷ Latner Thoracic Surgery Research Laboratories, Toronto General Hospital Research Institute, University Health Network, 101 College Street, PMCRT2-814, Toronto, ON, M5G 1L7, Canada. mingyao.liu@utoronto.ca.
⁸ Institute of Medical Science, Temerty Faculty of Medicine, University of Toronto, Toronto, ON, Canada. mingyao.liu@utoronto.ca.
⁹ Department of Surgery, Temerty Faculty of Medicine, University of Toronto, Toronto, ON, Canada. mingyao.liu@utoronto.ca.

^# Contributed equally.

Abstract

Background: Ischemia-reperfusion injury is a key complication following lung transplantation. The clinical application of ex vivo lung perfusion (EVLP) to assess donor lung function has significantly increased the utilization of "marginal" donor lungs with good clinical outcomes. The potential of EVLP on improving organ quality and ameliorating ischemia-reperfusion injury has been suggested.

Methods: To determine the effects of ischemia-reperfusion and EVLP on gene expression in human pulmonary microvascular endothelial cells and epithelial cells, cell culture models were used to simulate cold ischemia (4 °C for 18 h) followed by either warm reperfusion (DMEM + 10% FBS) or EVLP (acellular Steen solution) at 37 °C for 4 h. RNA samples were extracted for bulk RNA sequencing, and data were analyzed for significant differentially expressed genes and pathways.

Results: Endothelial and epithelial cells showed significant changes in gene expressions after ischemia-reperfusion or EVLP. Ischemia-reperfusion models of both cell types showed upregulated pro-inflammatory and downregulated cell metabolism pathways. EVLP models, on the other hand, exhibited downregulation of cell metabolism, without any inflammatory signals.

Conclusion: The commonly used acellular EVLP perfusate, Steen solution, silenced the activation of pro-inflammatory signaling in both human lung endothelial and epithelial cells, potentially through the lack of serum components. This finding could establish the basic groundwork of studying the benefits of EVLP perfusate as seen from current clinical practice.

Keywords: Cell culture model; Ex vivo organ perfusion; Ischemia reperfusion; Lung transplantation; RNA sequencing.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Endothelial Cells
Epithelial Cells
Humans
Ischemia
Lung / metabolism
Lung Transplantation* / adverse effects
Perfusion / adverse effects
Reperfusion Injury*

Grants and funding

PJT-148847/CIHR/Canada