scholarly journals Novel delivery of cellular therapy to reduce ischaemia reperfusion injury in kidney transplantation

2019 ◽  
Author(s):  
Emily R Thompson ◽  
Lucy Bates ◽  
Ibrahim K Ibrahim ◽  
Avinash Sewpaul ◽  
Ben Stenberg ◽  
...  
Keyword(s):  
Cell Therapy ◽  
Reperfusion Injury ◽  
Cellular Therapy ◽  
Ex Vivo ◽  
Left Kidney ◽  
Kidney Injury ◽  
Machine Perfusion ◽  
Ischaemia Reperfusion Injury ◽  
Ischaemia Reperfusion ◽  
Normothermic Machine Perfusion

AbstractEx-vivo normothermic machine perfusion (NMP) of donor kidneys prior to transplantation provides a platform for direct delivery of cellular therapeutics to optimise organ quality prior to transplantation. Multipotent Adult Progenitor Cells (MAPC®) possess potent immunomodulatory properties which could prove beneficial in minimising subsequent ischaemia reperfusion injury. We investigated the potential reconditioning capability of MAPC cells in kidney NMP.MethodsPairs (5) of human kidneys from the same donor were simultaneously perfused for 7 hours. The right or left kidney was randomly allocated to receive MAPC treatment. Serial samples of perfusate, urine and tissue biopsies were taken for comparison with the control paired kidney.ResultsMAPC-treated kidneys demonstrated improved urine output (p<0.01), decreased expression of the kidney injury biomarker NGAL (p<0.01), improved microvascular perfusion on contrast enhanced ultrasound (cortex p<0.05, medulla p<0.01), downregulation of IL-1β (p<0.05) and upregulation of IL-10 (p<0.05) and Indolamine-2, 3-dioxygenase (p<0.05). A mouse model of intraperitoneal chemotaxis demonstrated decreased neutrophil recruitment when stimulated with perfusate from MAPC-treated kidneys (p<0.01). Immunofluorescence revealed pre-labelled MAPC cells home to the perivascular space in the kidneys during NMP. MAPC therapy was not associated with detrimental physiological or embolic events.ConclusionWe report the first successful delivery of cellular therapy to a kidney during NMP. Kidneys treated with MAPC cells demonstrate improvement in clinically relevant functional parameters and injury biomarkers. This novel method of cell therapy delivery provides an exciting opportunity to recondition organs prior to clinical transplantation.One Sentence SummaryEx-vivo reconditioning of human kidneys using Multipotent Adult Progenitor Cell therapy delivered during normothermic machine perfusion.

scholarly journals Normothermic Machine Perfusion (NMP) of the Liver as a Platform for Therapeutic Interventions during Ex-Vivo Liver Preservation: A Review

2020 ◽  
Vol 9 (4) ◽  
pp. 1046 ◽  
Author(s):  
Fungai Dengu ◽  
Syed Hussain Abbas ◽  
Georg Ebeling ◽  
David Nasralla
Keyword(s):  
Ex Vivo ◽  
Organ Donor ◽  
Therapeutic Interventions ◽  
Machine Perfusion ◽  
Real Potential ◽  
Ischaemia Reperfusion Injury ◽  
Donor Pool ◽  
Liver Preservation ◽  
Ischaemia Reperfusion ◽  
Normothermic Machine Perfusion

Liver transplantation is increasingly dependent on the use of extended criteria donors (ECD) to increase the organ donor pool and address rising demand. This has necessitated the adoption of innovative technologies and strategies to protect these higher-risk grafts from the deleterious effects of traditional preservation and ischaemia reperfusion injury (IRI). The advent of normothermic machine perfusion (NMP) and rapid growth in the clinical adoption of this technology has accelerated efforts to utilise NMP as a platform for therapeutic intervention to optimise donor livers. In this review we will explore the emerging preclinical data related to ameliorating the effects of IRI, protecting the microcirculation and reducing the immunogenicity of donor organs during NMP. Exploiting the window of opportunity afforded by NMP, whereby the liver can be continuously supported and functionally assessed while therapies are directly delivered during the preservation period, has clear logistical and theoretical advantages over current preservation methods. The clinical translation of many of the therapeutic agents and strategies we will describe is becoming more feasible with widespread adaptation of NMP devices and rapid advances in molecular biology and gene therapy, which have substantially improved the performance of these agents. The delivery of novel therapeutics during NMP represents one of the new frontiers in transplantation research and offers real potential for successfully tackling fundamental challenges in transplantation such as IRI.

scholarly journals O25: TARGETING THE RENAL TUBULAR EPITHELIUM WITH ANTI-MIRNA THERAPY: A POTENTIAL MECHANISM FOR MINIMISING ISCHAEMIA REPERFUSION INJURY

2021 ◽  
Vol 108 (Supplement_1) ◽  
Author(s):  
E Irwin ◽  
E Thompson ◽  
S Tingle ◽  
P Ezuma ◽  
L Matthews ◽  
...  
Keyword(s):  
Reperfusion Injury ◽  
Cell Types ◽  
Specific Response ◽  
Machine Perfusion ◽  
Ischaemia Reperfusion Injury ◽  
Western Blots ◽  
Mrna Targets ◽  
Renal Tubular Epithelium ◽  
Ischaemia Reperfusion ◽  
Normothermic Machine Perfusion

Abstract Introduction Ischaemia reperfusion injury (IRI) is an unavoidable, significant consequence of renal transplantation. MicroRNAs are small, non-coding RNA molecules that regulate multiple downstream mRNA targets. MiRNA-21-5p and miRNA-24-3p have been previously implicated in IRI. Antisense oligonucleotides (ASOs) block specific microRNAs, with previous work by our group demonstrating their delivery to kidneys using normothermic machine perfusion. Imaging these kidneys revealed ASO localisation around proximal tubule epithelial cells (PTECs). This project aimed to characterise ASO blockade against miRNA-21-5p and miRNA-24-3p in PTECs. Method HKC8 cells, a human PTEC cell line, were used throughout these experiments. Cells were placed in a hypoxic incubator for 24hrs, followed by 6hrs of reoxygenation to mimic IRI. HKC8s were transfected with ASOs using lipofectamine. RT-qPCR and Western Blots were used to evaluate expression of antioxidant targets, SOD2 and HMOX1. Result MiRNA-21-5p and miRNA-24-3p levels were high throughout hypoxia and reoxygenation. Single blockade with anti-miRNA-21-5p resulted in a significant increase in its downstream target SOD2 (P&lt;0.05). Anti-miRNA-24-3p treatment resulted in no change in either of its downstream targets, HMOX1 or SOD2. This was reflected in the failure of dual blockade to produce a synergistic effect on the shared target, SOD2. Conclusion Anti-miRNA-21-5p results in a significant increase of SOD2, which is well characterised as protective during IRI. Anti-miRNA-24-3p appears to have no effect on PTECs, contrary to previous work in endothelial cells, perhaps suggesting a cell specific response of microRNAs. Normothermic machine perfusion could be used to deliver dual ASOs; allowing simultaneous targeting of different kidney cell types. Take-home message The delivery of anti-miRNA-21-5p therapy pre-transplant, using normothermic machine perfusion, has the potential to reduce ischaemia reperfusion injury and improve kidney transplant outcomes.

scholarly journals The IL-2/Anti-IL-2 Complex Attenuates Cardiac Ischaemia-Reperfusion Injury Through Expansion of Regulatory T Cells

2017 ◽  
Vol 44 (5) ◽  
pp. 1810-1827 ◽  
Author(s):  
Junhui Xiao ◽  
Kunwu Yu ◽  
Ming Li ◽  
Chuanyin Xiong ◽  
Yuzhen Wei ◽  
...  
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Reperfusion Injury ◽  
Sham Group ◽  
Myocardial Function ◽  
Ex Vivo ◽  
Inflammatory Factors ◽  
Ischaemia Reperfusion Injury ◽  
Ischaemia Reperfusion ◽  
Infarct Area

Background/Aims: Regulatory T cells (Tregs) can suppress immunologic damage in myocardial ischaemia/reperfusion injury (MIRI), however, the isolation and ex vivo expansion of these cells for clinical application remains challenging. Here, we investigated whether the IL-2/anti-IL-2 complex (IL-2C), a mediator of Treg expansion, can attenuate MIRI in mice. Methods: Myocardial I/R was surgically induced in male C57BL/6 mice, aged 8-10 weeks, that were randomly assigned to 1) sham group (Sham), 2) Phosphate Buffered Saline (PBS), 3) IL-2-anti-IL-2 Ab complex (IL-2C), or 4) sham group, 5) PBS, 6) IL-2C after MIRI, or 7) IL-2C, 8) IL-2C+anti-CD25 mAbs, or 9) IL-2C; 10) IL-2C+anti-TGF-β1 mAbs, 11) IL-2C+anti-IL-10 mAbs. The following parameters were measured at different time points: infarct area, myocardial apoptosis, splenocytes, the inhibitory function of Tregs, and presence of inflammatory factors. In addition, immunohistochemistry analysis was performed. Results: We observed that Tregs were activated in response to MIRI. IL-2C administered before MIRI induced Treg expansion in both spleen and heart, attenuated Th1 and Th17 cell numbers, improved myocardial function, and attenuated both infiltration of inflammatory cells and apoptosis after MIRI. Furthermore, IL-2C administration reduced expression of inflammatory cytokines in the heart and attenuated proliferation of splenic cells. Depletion of Tregs with anti-CD25 mAb abrogated the beneficial effects of IL-2C. However, IL-2C–mediated myocardial protection was not dependent on either IL-10 or TGF-β. In addition, IL-2C administration after MIRI did not reduce infarct area, but did improve myocardial function slightly and reduced myocardial fibrosis. Conclusion: Our results demonstrate that IL-2C–induced Treg expansion attenuates MIRI and improves myocardial recovery in vivo, suggesting that IL-2C is a promising therapeutic target for myocardial IRI.

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