MicroRNA-1 protects the endothelium in acute lung injury
Asawari Korde, Maria Haslip, Prachi Pednekar, Alamzeb Khan, Maurizio Chioccioli, Sameet Mehta, Francesc Lopez-Giraldez, Santos Bermejo, Mauricio Rojas, Charles Dela Cruz, Michael A. Matthay, Jordan S. Pober, Richard W. Pierce, Shervin S. Takyar
Asawari Korde, Maria Haslip, Prachi Pednekar, Alamzeb Khan, Maurizio Chioccioli, Sameet Mehta, Francesc Lopez-Giraldez, Santos Bermejo, Mauricio Rojas, Charles Dela Cruz, Michael A. Matthay, Jordan S. Pober, Richard W. Pierce, Shervin S. Takyar
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Research Article Pulmonology Vascular biology

MicroRNA-1 protects the endothelium in acute lung injury

Abstract

Acute lung injury (ALI) and its most severe form, acute respiratory distress syndrome (ARDS), cause severe endothelial dysfunction in the lung, and vascular endothelial growth factor (VEGF) is elevated in ARDS. We found that the levels of a VEGF-regulated microRNA, microRNA-1 (miR-1), were reduced in the lung endothelium after acute injury. Pulmonary endothelial cell–specific (EC-specific) overexpression of miR-1 protected the lung against cell death and barrier dysfunction in both murine and human models and increased the survival of mice after pneumonia-induced ALI. miR-1 had an intrinsic protective effect in pulmonary and other types of ECs; it inhibited apoptosis and necroptosis pathways and decreased capillary leak by protecting adherens and tight junctions. Comparative gene expression analysis and RISC recruitment assays identified miR-1 targets in the context of injury, including phosphodiesterase 5A (PDE5A), angiopoietin-2 (ANGPT2), CNKSR family member 3 (CNKSR3), and TNF-α–induced protein 2 (TNFAIP2). We validated miR-1–mediated regulation of ANGPT2 in both mouse and human ECs and found that in a 119-patient pneumonia cohort, miR-1 correlated inversely with ANGPT2. These findings illustrate a previously unknown role of miR-1 as a cytoprotective orchestrator of endothelial responses to acute injury with prognostic and therapeutic potential.

Authors

Asawari Korde, Maria Haslip, Prachi Pednekar, Alamzeb Khan, Maurizio Chioccioli, Sameet Mehta, Francesc Lopez-Giraldez, Santos Bermejo, Mauricio Rojas, Charles Dela Cruz, Michael A. Matthay, Jordan S. Pober, Richard W. Pierce, Shervin S. Takyar

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Figure 5

miR-1 protects human lungs against cell death.

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miR-1 protects human lungs against cell death. Human lung tissue samples...
Human lung tissue samples were cultured and transfected with a double-stranded miR-1 RNA mimic (miR-1) or control RNA (ctrl), followed by 24-hour treatment with LPS (500 ng/mL) or vehicle (PBS). (A) Percentage TUNEL-positive cells was measured and graphed as in Figure 4A (n = 3 patients, 6 replicates each). *P = 0.0016, **P = 0.0225, ***P = 0.0014, ****P = 0.0058. Scale bars: 100 μm. (B) LDH levels were measured in culture media, and the values were normalized to the mean of the PBS group (n = 3 patients, 2 replicates each). *P = 0.0067, **P = 0.0052, ***P = 0.0011, ****P = 0.0043. (C and D) PCLS samples from the human lung were cultured in the growth medium, transduced with V-miR-1 or V-Ctrl, and treated with LPS (500 ng/mL) for 24 hours. Paraffin-embedded sections were stained for TUNEL, von Willebrand factor (VWF, for endothelial cells), and epithelial cell adhesion molecule (EpCAM, for epithelial cells). (C) Representative confocal images (×200 magnification) of alveoli. The graph represents the quantification of percentage TUNEL-positive cells/length of the alveolar wall (n = 5 replicates for V-ctrl and 4 for V-miR-1). *P = 0.0004. (D) Representative images of small lung vessels. The graph represents percentage TUNEL-positive cells/perimeter of the vessel (n = 6 replicates per group). Scale bars: 50 μm (C and D). *P = 0.0002. Error bars represent the SEM. Data were analyzed by unpaired, 2-tailed t test with Welch’s correction or Mann-Whitney U test based on normality.