Expression of human thrombomodulin by GalTKO.hCD46 pigs modulates coagulation cascade activation by endothelial cells and during ex vivo lung perfusion with human blood.

IF 3.3 4区 医学 Q2 MEDICINE, RESEARCH & EXPERIMENTAL
Xenotransplantation Pub Date : 2023-11-01 Epub Date: 2023-09-28 DOI:10.1111/xen.12828
Lars Burdorf, Zhuo Gao, Andrea Riner, Evelyn Sievert, Donald G Harris, Kasinath V Kuravi, Benson H Morrill, Zahra Habibabady, Elana Rybak, Siamak Dahi, Tianshu Zhang, Evan Schwartz, Elizabeth Kang, Xiangfei Cheng, Charles T Esmon, Carol J Phelps, David L Ayares, Richard N Pierson Iii, Agnes M Azimzadeh
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引用次数: 0

Abstract

Thrombomodulin is important for the production of activated protein C (APC), a molecule with significant regulatory roles in coagulation and inflammation. To address known molecular incompatibilities between pig thrombomodulin and human thrombin that affect the conversion of protein C into APC, GalTKO.hCD46 pigs have been genetically modified to express human thrombomodulin (hTBM). The aim of this study was to evaluate the impact of transgenic hTBM expression on the coagulation dysregulation that is observed in association with lung xenograft injury in an established lung perfusion model, with and without additional blockade of nonphysiologic interactions between pig vWF and human GPIb axis. Expression of hTBM was variable between pigs at the transcriptional and protein level. hTBM increased the activation of human protein C and inhibited thrombosis in an in vitro flow perfusion assay, confirming that the expressed protein was functional. Decreased platelet activation was observed during ex vivo perfusion of GalTKO.hCD46 lungs expressing hTBM and, in conjunction with transgenic hTBM, blockade of the platelet GPIb receptor further inhibited platelets and increased survival time. Altogether, our data indicate that expression of transgenic hTBM partially addresses coagulation pathway dysregulation associated with pig lung xenograft injury and, in combination with vWF-GP1b-directed strategies, is a promising approach to improve the outcomes of lung xenotransplantation.

GalTKO.hCD46猪对人血栓调节蛋白的表达调节内皮细胞的凝血级联激活和人血离体肺灌注过程。
血栓调节蛋白对活化蛋白C(APC)的产生很重要,APC是一种在凝血和炎症中具有重要调节作用的分子。为了解决影响蛋白质C转化为APC的猪血栓调节蛋白和人凝血酶之间已知的分子不相容性,对GalTKO.hCD46猪进行了基因修饰,以表达人血栓调节蛋白(hTBM)。本研究的目的是评估转基因hTBM表达对凝血失调的影响,在已建立的肺灌注模型中,观察到与肺异种移植物损伤相关的凝血失调,在有或没有额外阻断猪vWF和人GPIb轴之间的非生理相互作用的情况下。hTBM的表达在转录和蛋白质水平上在猪之间是可变的。hTBM在体外流动灌注试验中增加了人蛋白C的激活并抑制了血栓形成,证实了所表达的蛋白是功能性的。在表达hTBM的GalTKO.hCD46肺的离体灌注过程中观察到血小板活化减少,并且与转基因hTBM结合,阻断血小板GPIb受体进一步抑制血小板并增加存活时间。总之,我们的数据表明,转基因hTBM的表达部分解决了与猪肺异种移植物损伤相关的凝血途径失调问题,并且与vWF-GP1b指导的策略相结合,是改善异种肺移植结果的一种有前途的方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Xenotransplantation
Xenotransplantation 医学-医学:研究与实验
CiteScore
6.80
自引率
15.40%
发文量
58
审稿时长
>12 weeks
期刊介绍: Xenotransplantation provides its readership with rapid communication of new findings in the field of organ and tissue transplantation across species barriers.The journal is not only of interest to those whose primary area is xenotransplantation, but also to veterinarians, microbiologists and geneticists. It also investigates and reports on the controversial theological, ethical, legal and psychological implications of xenotransplantation.
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