Habertheuer Andreas, Ram Chirag, Schmierer Maggie, Chatterjee Shampa, Hu Robert, Freas Andrew, Zielinski Patrick, Rogers Wade, Silvestro Eva M, McGrane Michael, Moore Jonni S, Korutla Laxminarayana, Siddiqui Sarmad, Xin Yi, Rizi Rahim, Qin Tao Jian, Kreisel Daniel, Naji Ali, Ochiya Takahiro, Vallabhajosyula Prashanth
Division of Cardiovascular Surgery, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA.
Division of Cardiac Surgery, Brigham and Women's Hospital, Harvard Medical School, Boston, MA.
Transplantation. 2022 Apr 1;106(4):754-766. doi: 10.1097/TP.0000000000003820.
There is a critical need for development of biomarkers to noninvasively monitor for lung transplant rejection. We investigated the potential of circulating donor lung-specific exosome profiles for time-sensitive diagnosis of acute rejection in a rat orthotopic lung transplant model.
Left lungs from Wistar transgenic rats expressing human CD63-GFP, an exosome marker, were transplanted into fully MHC-mismatched Lewis recipients or syngeneic controls. Recipient blood was collected between 4 h and 10 d after transplantation, and plasma was processed for exosome isolation by size exclusion column chromatography and ultracentrifugation. Circulating donor exosomes were profiled using antihuman CD63 antibody quantum dot on the nanoparticle detector and via GFP trigger on the nanoparticle flow cytometer.
In syngeneic controls, steady-state levels of circulating donor exosomes were detected at all posttransplant time points. Allogeneic grafts lost perfusion by day 8, consistent with acute rejection. Levels of circulating donor exosomes peaked on day 1, decreased significantly by day 2, and then reached baseline levels by day 3. Notably, decrease in peripheral donor exosome levels occurred before grafts had histological evidence of acute rejection.
Circulating donor lung-specific exosome profiles enable an early detection of acute rejection before histologic manifestation of injury to the pulmonary allograft. As acute rejection episodes are a major risk factor for the development of chronic lung allograft dysfunction, this biomarker may provide a novel noninvasive diagnostic platform that can translate into earlier therapeutic intervention for lung transplant patients.
迫切需要开发生物标志物以无创监测肺移植排斥反应。我们在大鼠原位肺移植模型中研究了循环供体肺特异性外泌体谱对急性排斥反应进行时间敏感性诊断的潜力。
将表达人CD63 - GFP（一种外泌体标志物）的Wistar转基因大鼠的左肺移植到完全MHC不匹配的Lewis受体或同基因对照中。在移植后4小时至10天收集受体血液，通过尺寸排阻柱色谱法和超速离心法处理血浆以分离外泌体。使用抗人CD63抗体量子点在纳米颗粒检测器上以及通过纳米颗粒流式细胞仪上的GFP触发对循环供体外泌体进行分析。
在同基因对照中，在所有移植后时间点均检测到循环供体外泌体的稳态水平。异基因移植物在第8天失去灌注，与急性排斥反应一致。循环供体外泌体水平在第1天达到峰值，在第2天显著下降，然后在第3天达到基线水平。值得注意的是，外周供体外泌体水平的下降发生在移植物出现急性排斥反应的组织学证据之前。
循环供体肺特异性外泌体谱能够在肺同种异体移植损伤的组织学表现之前早期检测急性排斥反应。由于急性排斥反应是慢性肺同种异体移植功能障碍发展的主要危险因素，这种生物标志物可能提供一种新型的无创诊断平台，可转化为对肺移植患者的早期治疗干预。
