tetano
Editor, Senior Moderator
Pulm Circ
. 2022 May 11;12(2):e12071.
doi: 10.1002/pul2.12071. eCollection 2022 Apr.
Pulmonary endothelial NEDD9 and the prothrombotic pathophenotype of acute respiratory distress syndrome due to SARS-CoV-2 infection
George A Alba[SUP] 1 [/SUP], Andriy O Samokhin[SUP] 2 [/SUP], Rui-Sheng Wang[SUP] 2 [/SUP], Bradley M Wertheim[SUP] 3 [/SUP], Kathleen J Haley[SUP] 3 [/SUP], Robert F Padera[SUP] 4 [/SUP], Sara O Vargas[SUP] 5 [/SUP], Ivan O Rosas[SUP] 6 [/SUP], Lida P Hariri[SUP] 1 7 [/SUP], Angela Shih[SUP] 7 [/SUP], Boyd Taylor Thompson[SUP] 1 [/SUP], Richard N Mitchell[SUP] 4 [/SUP], Bradley A Maron[SUP] 2 [/SUP]
Affiliations
Abstract
The pathobiology of in situ pulmonary thrombosis in acute respiratory distress syndrome (ARDS) due to severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection is incompletely characterized. In human pulmonary artery endothelial cells (HPAECs), hypoxia increases neural precursor cell expressed, developmentally downregulated 9 (NEDD9) and induces expression of a prothrombotic NEDD9 peptide (N9[SUB]P[/SUB]) on the extracellular plasma membrane surface. We hypothesized that the SARS-CoV-2-ARDS pathophenotype involves increased pulmonary endothelial N9[SUB]P[/SUB]. Paraffin-embedded autopsy lung specimens were acquired from patients with SARS-CoV-2-ARDS (n = 13), ARDS from other causes (n = 10), and organ donor controls (n = 5). Immunofluorescence characterized the expression of N9[SUB]P[/SUB], fibrin, and transcription factor 12 (TCF12), a putative binding target of SARS-CoV-2 and known transcriptional regulator of NEDD9. We performed RNA-sequencing on normal HPAECs treated with normoxia or hypoxia (0.2% O[SUB]2[/SUB]) for 24 h. Immunoprecipitation-liquid chromatography-mass spectrometry (IP-LC-MS) profiled protein-protein interactions involving N9[SUB]P[/SUB] relevant to thrombus stabilization. Hypoxia increased TCF12 messenger RNA significantly compared to normoxia in HPAECs in vitro (+1.19-fold, p = 0.001; false discovery rate = 0.005), and pulmonary endothelial TCF12 expression was increased threefold in SARS-CoV-2-ARDS versus donor control lungs (p < 0.001). Compared to donor controls, pulmonary endothelial N9[SUB]P[/SUB]-fibrin colocalization was increased in situ in non-SARS-CoV-2-ARDS and SARS-CoV-2-ARDS decedents (3.7 ± 1.2 vs. 10.3 ± 3.2 and 21.8 ± 4.0 arb. units, p < 0.001). However, total pulmonary endothelial N9[SUB]P[/SUB] was increased significantly only in SARS-CoV-2-ARDS versus donor controls (15 ± 4.2 vs. 6.3 ± 0.9 arb. units, p < 0.001). In HPAEC plasma membrane isolates, IP-LC-MS identified a novel protein-protein interaction between NEDD9 and the β[SUB]3[/SUB]-subunit of the α[SUB]v[/SUB]β[SUB]3[/SUB]-integrin, which regulates fibrin anchoring to endothelial cells. In conclusion, lethal SARS-CoV-2-ARDS is associated with increased pulmonary endothelial N9[SUB]P[/SUB] expression and N9[SUB]P[/SUB]-fibrin colocalization in situ. Further investigation is needed to determine the pathogenetic and potential therapeutic relevance of N9[SUB]P[/SUB] to the thrombotic pathophenotype of SARS-CoV-2-ARDS.
Keywords: SARS‐CoV‐2; acute respiratory distress syndrome; endothelium; pulmonary biology; thrombosis.
. 2022 May 11;12(2):e12071.
doi: 10.1002/pul2.12071. eCollection 2022 Apr.
Pulmonary endothelial NEDD9 and the prothrombotic pathophenotype of acute respiratory distress syndrome due to SARS-CoV-2 infection
George A Alba[SUP] 1 [/SUP], Andriy O Samokhin[SUP] 2 [/SUP], Rui-Sheng Wang[SUP] 2 [/SUP], Bradley M Wertheim[SUP] 3 [/SUP], Kathleen J Haley[SUP] 3 [/SUP], Robert F Padera[SUP] 4 [/SUP], Sara O Vargas[SUP] 5 [/SUP], Ivan O Rosas[SUP] 6 [/SUP], Lida P Hariri[SUP] 1 7 [/SUP], Angela Shih[SUP] 7 [/SUP], Boyd Taylor Thompson[SUP] 1 [/SUP], Richard N Mitchell[SUP] 4 [/SUP], Bradley A Maron[SUP] 2 [/SUP]
Affiliations
- PMID: 35599981
- PMCID: PMC9111030
- DOI: 10.1002/pul2.12071
Abstract
The pathobiology of in situ pulmonary thrombosis in acute respiratory distress syndrome (ARDS) due to severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection is incompletely characterized. In human pulmonary artery endothelial cells (HPAECs), hypoxia increases neural precursor cell expressed, developmentally downregulated 9 (NEDD9) and induces expression of a prothrombotic NEDD9 peptide (N9[SUB]P[/SUB]) on the extracellular plasma membrane surface. We hypothesized that the SARS-CoV-2-ARDS pathophenotype involves increased pulmonary endothelial N9[SUB]P[/SUB]. Paraffin-embedded autopsy lung specimens were acquired from patients with SARS-CoV-2-ARDS (n = 13), ARDS from other causes (n = 10), and organ donor controls (n = 5). Immunofluorescence characterized the expression of N9[SUB]P[/SUB], fibrin, and transcription factor 12 (TCF12), a putative binding target of SARS-CoV-2 and known transcriptional regulator of NEDD9. We performed RNA-sequencing on normal HPAECs treated with normoxia or hypoxia (0.2% O[SUB]2[/SUB]) for 24 h. Immunoprecipitation-liquid chromatography-mass spectrometry (IP-LC-MS) profiled protein-protein interactions involving N9[SUB]P[/SUB] relevant to thrombus stabilization. Hypoxia increased TCF12 messenger RNA significantly compared to normoxia in HPAECs in vitro (+1.19-fold, p = 0.001; false discovery rate = 0.005), and pulmonary endothelial TCF12 expression was increased threefold in SARS-CoV-2-ARDS versus donor control lungs (p < 0.001). Compared to donor controls, pulmonary endothelial N9[SUB]P[/SUB]-fibrin colocalization was increased in situ in non-SARS-CoV-2-ARDS and SARS-CoV-2-ARDS decedents (3.7 ± 1.2 vs. 10.3 ± 3.2 and 21.8 ± 4.0 arb. units, p < 0.001). However, total pulmonary endothelial N9[SUB]P[/SUB] was increased significantly only in SARS-CoV-2-ARDS versus donor controls (15 ± 4.2 vs. 6.3 ± 0.9 arb. units, p < 0.001). In HPAEC plasma membrane isolates, IP-LC-MS identified a novel protein-protein interaction between NEDD9 and the β[SUB]3[/SUB]-subunit of the α[SUB]v[/SUB]β[SUB]3[/SUB]-integrin, which regulates fibrin anchoring to endothelial cells. In conclusion, lethal SARS-CoV-2-ARDS is associated with increased pulmonary endothelial N9[SUB]P[/SUB] expression and N9[SUB]P[/SUB]-fibrin colocalization in situ. Further investigation is needed to determine the pathogenetic and potential therapeutic relevance of N9[SUB]P[/SUB] to the thrombotic pathophenotype of SARS-CoV-2-ARDS.
Keywords: SARS‐CoV‐2; acute respiratory distress syndrome; endothelium; pulmonary biology; thrombosis.