Ex vivo lung perfusion as a human platform for preclinical small molecule testing

Weathington, Nathaniel M.; Álvarez, Diana; Sembrat, John; Radder, Josiah E.; Cárdenes, Nayra; Noda, Kentaro; Gong, Qiaoke; Wong, Hesper; Kolls, Jay K.; D’Cunha, Jonathan; Mallampalli, Rama K.; Chen, Bill B.; Rojas, Mauricio

doi:10.1172/jci.insight.95515

Cited by 26 publications

(26 citation statements)

References 30 publications

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“…Several groups have demonstrated the feasibility of treating donor lungs with pulmonary emboli using urokinase or alteplase during EVLP before successful transplantation (52,53). In addition, several groups have used EVLP systems to study the mechanisms of primary graft dysfunction (54) and to test novel therapeutics (50,55).…”

Section: Clinical Applications Of Ex Vivo Lung Perfusionmentioning

confidence: 99%

The ex vivo human lung: research value for translational science

Ross¹,

Nesseler²,

Lee³

et al. 2019

JCI Insight

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Section: Clinical Applications Of Ex Vivo Lung Perfusionmentioning

confidence: 99%

The ex vivo human lung: research value for translational science

Ross¹,

Nesseler²,

Lee³

et al. 2019

JCI Insight

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“…The experimental preparation has improved the available pool of lungs appropriate for transplantation [4][5][6][7][8][9] and understanding mechanisms that contribute to primary graft dysfunction in lung transplant recipients [10]. It also offers insight into lung physiology in injury [11,12] and as such, allows for the testing of new therapeutics (mesenchymal stem cells, microvesicles) [13][14][15][16].…”

Section: Introductionmentioning

confidence: 99%

The endogenous capacity to produce proinflammatory mediators by the ex vivo human perfused lung

Leligdowicz

Ross

Nesseler

et al. 2020

ICMx

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Background The ex vivo human perfused lung model has enabled optimizing donor lungs for transplantation and delineating mechanisms of lung injury. Perfusate and airspace biomarkers are a proxy of the lung response to experimental conditions. However, there is a lack of studies evaluating biomarker kinetics during perfusion and after exposure to stimuli. In this study, we analyzed the ex vivo-perfused lung response to three key perturbations: exposure to the perfusion circuit, exogenous fresh whole blood, and bacteria. Results Ninety-nine lungs rejected for transplantation underwent ex vivo perfusion. One hour after reaching experimental conditions, fresh whole blood was added to the perfusate (n = 55). Two hours after reaching target temperature, Streptococcus pneumoniae was added to the perfusate (n = 42) or to the airspaces (n = 17). Perfusate and airspace samples were collected at baseline (once lungs were equilibrated for 1 h, but before blood or bacteria were added) and 4 h later. Interleukin (IL)-6, IL-8, angiopoietin (Ang)-2, and soluble tumor necrosis factor receptor (sTNFR)-1 were quantified. Baseline perfusate and airspace biomarker levels varied significantly, and this was not related to pre-procurement PaO2:FiO2 ratio, cold ischemia time, and baseline alveolar fluid clearance (AFC). After 4 h of ex vivo perfusion, the lung demonstrated a sustained production of proinflammatory mediators. The change in biomarker levels was not influenced by baseline donor lung characteristics (cold ischemia time, baseline AFC) nor was it associated with measures of experimental epithelial (final AFC) or endothelial (percent weight gain) injury. In the presence of exogenous blood, the rise in biomarkers was attenuated. Lungs exposed to intravenous (IV) bacteria relative to control lungs demonstrated a significantly higher rise in perfusate IL-6. Conclusions The ex vivo-perfused lung has a marked endogenous capacity to produce inflammatory mediators over the course of short-term perfusion that is not significantly influenced by donor lung characteristics or the presence of exogenous blood, and only minimally affected by the introduction of systemic bacteremia. The lack of association between biomarker change and donor lung cold ischemia time, final alveolar fluid clearance, and experimental percent weight gain suggests that the maintained ability of the human lung to produce biomarkers is not merely a marker of lung epithelial or endothelial injury, but may support the function of the lung as an immune cell reservoir.

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“…The impact of this change in the cytokine secretion pattern in the transplantation outcome needs to be investigated in detail in future experiments. Currently, different approaches to prevent the release of pro-inflammatory cytokines by ex vivo perfused organs based on the use of membranes or small molecules are being developed to avoid tissue impairment during perfusion or the activation and polarization of immune responses after transplantation (54,55).…”

Section: Immuno-engineering Of the Kidneymentioning

confidence: 99%

Genetic Engineering of the Kidney to Permanently Silence MHC Transcripts During ex vivo Organ Perfusion

et al. 2020

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Organ gene therapy represents a promising tool to correct diseases or improve graft survival after transplantation. Polymorphic variation of the major histocompatibility complex (MHC) antigens remains a major obstacle to long-term graft survival after transplantation. Previously, we demonstrated that MHC-silenced cells are protected against allogeneic immune responses. We also showed the feasibility to silence MHC in the lung. Here, we aimed at the genetic engineering of the kidney toward permanent silencing of MHC antigens in a rat model. We constructed a sub-normothermic ex vivo perfusion system to deliver lentiviral vectors encoding shRNAs targeting β2-microglobulin and the class II transactivator to the kidney. In addition, the vector contained the sequence for a secreted nanoluciferase. After kidney transplantation (ktx), we detected bioluminescence in the plasma and urine of recipients of an engineered kidney during the 6 weeks of post-transplant monitoring, indicating a stable transgene expression. Remarkably, transcript levels of β2-microglobulin and the class II transactivator were decreased by 70% in kidneys expressing specific shRNAs. Kidney genetic modification did not cause additional cell death compared to control kidneys after machine perfusion. Nevertheless, cytokine secretion signatures were altered during perfusion with lentiviral vectors as revealed by an increase in the secretion of IL-10, MIP-1α, MIP-2, IP-10, and EGF and a decrease in the levels of IL-12, IL-17, MCP-1, and IFN-γ. Biodistribution assays indicate that the localization of the vector was restricted to the graft. This study shows the potential to generate immunologically invisible kidneys showing great promise to support graft survival after transplantation and may contribute to reduce the burden of immunosuppression.

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Ex vivo lung perfusion as a human platform for preclinical small molecule testing

Cited by 26 publications

References 30 publications

The ex vivo human lung: research value for translational science

The ex vivo human lung: research value for translational science

The endogenous capacity to produce proinflammatory mediators by the ex vivo human perfused lung

Genetic Engineering of the Kidney to Permanently Silence MHC Transcripts During ex vivo Organ Perfusion

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