Dynamic conditioning of porcine kidney grafts with extracellular vesicles derived from urine progenitor cells: A proof-of-concept study

IF 7.9 1区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Clinical and Translational Medicine Pub Date : 2024-12-13 DOI:10.1002/ctm2.70095

Perrine Burdeyron, Sébastien Giraud, Maryne Lepoittevin, Nina Jordan, Sonia Brishoual, Maïté Jacquard, Virginie Ameteau, Nadège Boildieu, Estelle Lemarie, Jonathan Daniel, Frédéric Martins, Nicolas Mélis, Marine Coué, Raphaël Thuillier, Henri Leuvenink, Luc Pellerin, Thierry Hauet, Clara Steichen

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引用次数: 0

Abstract

Among strategies to limit ischemia/reperfusion (IR) injuries in transplantation, cell therapy using stem cells to condition/repair transplanted organs appears promising. We hypothesized that using a cell therapy based on extracellular vesicles (EVs) derived from urine progenitor cells (UPCs) during hypothermic and normothermic machine perfusion can prevent IR-related kidney damage.

We isolated and characterized porcine UPCs and their extracellular vesicles (EVs). Then these were used in an ex vivo porcine kidney preservation model. Kidneys were subjected to warm ischemia (32 min) and then preserved by hypothermic machine perfusion (HMP) for 24 h before 5 h of normothermic machine perfusion (NMP). Three groups were performed (n = 5–6): Group 1 (G1): HMP/vehicle + NMP/vehicle, Group 2 (G2): HMP/EVs + NMP/vehicle, Group 3 (G3): HMP/EVs + NMP/EVs.

Porcine UPCs were successfully isolated from urine and fully characterized as well as their EVs which were found of expected size/phenotype. EVs injection during HMP alone, NMP alone, or both was feasible and safe and did not impact perfusion parameters. However, cell damage markers (LDH, ASAT) were decreased in G3 compared with G1, and G3 kidneys displayed a preserved tissue integrity with reduced tubular dilatation and inflammation notably. However, renal function indicators such as creatinine clearance measured for 5 h of normothermic perfusion or NGAL perfusate's level were not modified by EVs injection. Regarding perfusate analysis, metabolomic analyses and cytokine quantification showed an immunomodulation signature in G3 compared with G1 and highlighted potential metabolic targets. In vitro, EVs as well as perfusates from G3 partially recovered endothelial cell metabolic activity after hypoxia. Finally, RNA-seq performed on kidney biopsies showed different profiles between G1 and G3 with regulation of potential IR targets of EVs therapy.

We showed the feasibility/efficacy of UPC-EVs for hypothermic/normothermic kidney conditioning before transplantation, paving the way for combining machine perfusion with EVs-based cell therapy for organ conditioning.

Highlights

· UPCs from porcine urine can be used to generate a cell therapy product based on extracellular vesicles (pUPC-EVs).
· pUPC-EVs injection during HMP and NMP decreases cell damage markers and has an immunomodulatory effect.
· pUPC-EVs-treated kidneys have distinct biochemical, metabolic, and transcriptomic profiles highlighting targets of interest.
· Our results pave the way for combining machine perfusion with EV-based cell therapy for kidney conditioning.

Abstract Image

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基于尿祖细胞的细胞外囊泡的猪肾移植物的动态调节：一项概念验证研究。

在限制移植中缺血/再灌注（IR）损伤的策略中，使用干细胞调节/修复移植器官的细胞治疗看起来很有希望。我们假设，在低温和常温机器灌注期间，使用基于源自尿祖细胞（UPCs）的细胞外囊泡（ev）的细胞疗法可以预防ir相关的肾脏损伤。我们分离并鉴定了猪UPCs及其细胞外囊泡（EVs）。然后将其用于离体猪肾保存模型。取肾脏进行热缺血（32 min）后，低温机器灌注（HMP）保存24 h，再进行恒温机器灌注（NMP） 5 h。分为3组（n = 5-6）：第1组（G1）： HMP/vehicle + NMP/vehicle，第2组（G2）： HMP/ ev + NMP/vehicle，第3组（G3）： HMP/ ev + NMP/ ev。成功地从猪尿液中分离出UPCs，并对其进行了充分的表征，并发现其ev符合预期的大小/表型。单独HMP、单独NMP或两者同时注射EVs是可行和安全的，且不影响灌注参数。然而，与G1相比，G3的细胞损伤标志物（LDH， ASAT）降低，G3的肾脏显示出组织完整性，肾小管扩张和炎症明显减少。然而，正常灌注5h时的肾功能指标，如肌酐清除率或灌注NGAL水平均未被EVs改变。在灌注分析方面，代谢组学分析和细胞因子定量显示G3与G1相比具有免疫调节特征，并突出了潜在的代谢靶点。体外缺氧后，内皮细胞和G3灌注液部分恢复内皮细胞代谢活性。最后，在肾活检中进行的RNA-seq显示G1和G3之间的不同特征与ev治疗的潜在IR靶点的调节。我们证明了upc - ev用于移植前低温/常温肾脏调节的可行性/有效性，为机器灌注与ev细胞治疗相结合进行器官调节铺平了道路。·猪尿中的UPCs可用于生产基于细胞外囊泡（pupc - ev）的细胞治疗产品。·在HMP和NMP期间注射pupc - ev可降低细胞损伤标志物，具有免疫调节作用。·pupc - ev处理的肾脏具有不同的生化、代谢和转录组谱，突出了感兴趣的靶点。·我们的研究结果为将机器灌注与基于ev的细胞治疗相结合用于肾脏调节铺平了道路。

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来源期刊

Clinical and Translational Medicine Multiple-

CiteScore

15.90

自引率

1.90%

发文量

450

审稿时长

4 weeks

期刊介绍： Clinical and Translational Medicine (CTM) is an international, peer-reviewed, open-access journal dedicated to accelerating the translation of preclinical research into clinical applications and fostering communication between basic and clinical scientists. It highlights the clinical potential and application of various fields including biotechnologies, biomaterials, bioengineering, biomarkers, molecular medicine, omics science, bioinformatics, immunology, molecular imaging, drug discovery, regulation, and health policy. With a focus on the bench-to-bedside approach, CTM prioritizes studies and clinical observations that generate hypotheses relevant to patients and diseases, guiding investigations in cellular and molecular medicine. The journal encourages submissions from clinicians, researchers, policymakers, and industry professionals.