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CC BY-NC-ND 4.0 · Thromb Haemost 2024; 124(07): 656-668
DOI: 10.1055/s-0044-1782182
Blood Cells, Inflammation and Infection

Endothelial LAT1 (SLC7A5) Mediates S-Nitrosothiol Import and Modulates Respiratory Sequelae of Red Blood Cell Transfusion In Vivo

Hongmei Zhu
1   Department of Medicine, Duke University Medical Center, Durham, North Carolina, United States
2   Durham VA Health Care System, Durham North Carolina, United States
,
Richard L. Auten
3   Department of Pediatrics, Duke University Medical Center, Durham, North Carolina, United States
,
Augustus Richard Whorton
4   Department of Pharmacology, Duke University Medical Center, Durham, North Carolina, United States
,
Stanley Nicholas Mason
3   Department of Pediatrics, Duke University Medical Center, Durham, North Carolina, United States
,
Cheryl B. Bock
5   Rodent Cancer Models Shared Resource, Duke University Medical Center, Durham, North Carolina, United States
,
Gary T. Kucera
5   Rodent Cancer Models Shared Resource, Duke University Medical Center, Durham, North Carolina, United States
,
Zachary T. Kelleher
1   Department of Medicine, Duke University Medical Center, Durham, North Carolina, United States
,
Aaron T. Vose
1   Department of Medicine, Duke University Medical Center, Durham, North Carolina, United States
2   Durham VA Health Care System, Durham North Carolina, United States
,
Tim J. McMahon 
1   Department of Medicine, Duke University Medical Center, Durham, North Carolina, United States
2   Durham VA Health Care System, Durham North Carolina, United States
› Author Affiliations Funding Veterans Administration (BX-003478) and NIH (P30 CA-014236 and R01 HL-161071).
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Abstract

Background Increased adhesivity of red blood cells (RBCs) to endothelial cells (ECs) may contribute to organ dysfunction in malaria, sickle cell disease, and diabetes. RBCs normally export nitric oxide (NO)-derived vascular signals, facilitating blood flow. S-nitrosothiols (SNOs) are thiol adducts formed in RBCs from precursor NO upon the oxygenation-linked allosteric transition in hemoglobin. RBCs export these vasoregulatory SNOs on demand, thereby regulating regional blood flow and preventing RBC–EC adhesion, and the large (system L) neutral amino acid transporter 1 (LAT1; SLC7A5) appears to mediate SNO export by RBCs.

Methods To determine the role of LAT1-mediated SNO import by ECs generally and of LAT1-mediated SNO import by ECs in RBC SNO-dependent modulation of RBC sequestration and blood oxygenation in vivo, we engineered LAT1fl/fl; Cdh5-Cre+ mice, in which the putative SNO transporter LAT1 can be inducibly depleted (knocked down, KD) specifically in ECs (“LAT1ECKD”).

Results We show that LAT1 in mouse lung ECs mediates cellular SNO uptake. ECs from LAT1ECKD mice (tamoxifen-induced LAT1fl/fl; Cdh5-Cre+) import SNOs poorly ex vivo compared with ECs from wild-type (tamoxifen-treated LAT1fl/fl; Cdh5-Cre−) mice. In vivo, endothelial depletion of LAT1 increased RBC sequestration in the lung and decreased blood oxygenation after RBC transfusion.

Conclusion This is the first study showing a role for SNO transport by LAT1 in ECs in a genetic mouse model. We provide the first direct evidence for the coordination of RBC SNO export with EC SNO import via LAT1. SNO flux via LAT1 modulates RBC–EC sequestration in lungs after transfusion, and its disruption impairs blood oxygenation by the lung.

Keywords

endothelial cells - S-nitrosothiols - adhesion - erythrocyte - respiratory

Supplementary Material



Publication History

Received: 15 March 2023

Accepted: 03 January 2024

Article published online:
22 March 2024

© 2024. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/)

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