Incorporating hepatic cell function into lung ex vivo lung perfusion for transplant preservation

将肝细胞功能纳入肺离体肺灌注以保存移植物

基本信息

批准号：
10666953
负责人：
JOERG C. GERLACH
金额：
$ 23.85万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10666953
关键词：
Address Biochemical Biological Biomedical Engineering Bioreactors Blood specimen Cadaver Cell physiology Cells Chronic Circulation Conditioned Culture Media Cross Circulation Cryopreservation Cytokine Signaling Data Drug Metabolic Detoxication Endothelium Experimental Models Family suidae Genetic Genetic Transcription Hepatic Hepatocyte Histologic Hour Human In Vitro Individual Inflammation Inflammatory Injury Intervention Investigation Ischemia Lung Lung Transplantation Measures Metabolism Mitochondria Modeling National Heart, Lung, and Blood Institute Nuclear Organ Organ Donor Outcome Patients Perfusion Phenotype Play Rattus Reaction Regulation Research Resuscitation Risk Risk Factors Role Running Signal Pathway Signal Transduction Structure of parenchyma of lung Survival Rate Technology Testing Time Tissue Preservation Tissue Transplantation Tissue Viability Tissues Transplant Recipients Transplantation Transportation United States National Institutes of Health Vascular Endothelium Waiting Lists Work cell injury clinically significant cytokine experimental study graft dysfunction human model improved in vitro testing in vivo in vivo evaluation inhibitor liver function liver injury lung preservation mortality pharmacologic post-transplant preservation pulmonary function receptor for advanced glycation endproducts response transplant model usability ventilation

项目摘要

ABSTRACT RESUBMISSION 1R21 HL164436-01 IN RESPONSE TO NHLBI “NOT-HL-21-024 BOLD AND NEW BIOENGINEERING RESEARCH” In contrast with other organs, preservation times of lung grafts for transplantation are limited to no more than 8 hours with standard cold preservation. Declining quality with extending preservation times increases the risk of primary graft dysfunction, a risk factor for developing chronic rejection, which could account for the low 5-year survival rate (50%) following lung transplantation (LTx). Ex Vivo Lung Perfusion (EVLP) is expected to improve lung preservation and transplant outcomes by providing normothermic circulation and ventilation. A problem is that current EVLP is self-limited by normothermia-activated lung metabolism. The accumulation of toxic metabolites leads to a proinflammatory state, activating Receptor of Advanced Glycation End-Products (RAGE) and nuclear factor (NF)-B mechanisms, which in turn upregulate proinflammatory cytokine signaling. Experimentally, cross-circulation of a whole swine with EVLP by others demonstrated improved lung resuscitation, but the exact contributors to this improvement remain unclear. Own in vitro preliminary data suggest a role for hepatic function in enhancing endothelial preservation. In previous work, we have maintained normal hepatic detoxification, synthesis, and regulation in in vitro circuits using liver cell bioreactors (BRx). We hypothesize first that the liver function in the swine cross-circulation model played a major role in enhancing EVLP and tissue viability. We also hypothesize that a hepatocyte BRx can partly substitute for whole-swine liver function on EVLP and maintain the observed improvements lung preservation and LTx outcomes. Our Specific Aim is to provide proof-of-principle for the ability of hepatocyte BRx to enhance lung graft preservation. We will incorporate a hepatic BRx in our established EVLP circuits and demonstrate its effect on short-term LTx in experimental rat models. We will repeat these experiments using a cadaveric human EVLP model. We will conduct comprehensive phenotypic, transcriptional, and functional endpoint assessments on lung tissue and hepatic cells in BRx, such as RAGE and NF-B. If successful, our technology will provide the means to change the current state of EVLP for lung preservation and allow lungs to be maintained longer outside the donor body with less cellular injury. Ultimately, our work will address current limitations in LTx, including increasing viable donor organ transportation time and distance, helping reduce LTx waiting lists, and improving post-transplant patient survival.

摘要重新提交 1R21 HL164436-01 回应 NHLBI“NOT-HL-21-024 大胆而新颖的生物工程研究” 与其他器官相比，用于移植的肺移植物的保存时间有限标准冷藏不超过8小时，随时间延长品质下降。保存时间会增加原发性移植物功能障碍的风险，这是发展的危险因素慢性排斥反应，这可能是肺移植术后 5 年生存率较低 (50%) 的原因移植（LTx）。离体肺灌注（EVLP）有望改善肺保存。通过提供常温循环和通气来改善移植结果一个问题是目前的EVLP是通过常温激活的肺代谢进行自我限制的。有毒代谢物导致促炎状态，激活高级糖化受体最终产物 (RAGE) 和核因子 (NF)-β 机制，进而上调实验上，整头猪与 EVLP 的交叉循环。其他人证明了肺复苏的改善，但具体的贡献者自己的体外初步数据表明肝功能的作用仍不清楚。在之前的工作中，我们维持了正常的肝脏。使用肝细胞生物反应器 (BRx) 进行体外回路解毒、合成和调节。首先捕捉到肝功能在猪交叉循环模型中发挥了主要作用我们还认为，肝细胞 BRx 可以部分增强 EVLP 和组织活力。替代 EVLP 上的全猪肝功能并维持观察到的肺部改善我们的具体目标是为该能力提供原理证明。肝细胞 BRx 以增强肺移植物的保存我们将在我们的研究中纳入肝 BRx。建立 EVLP 电路并证明其对实验大鼠短期 LTx 的影响我们将使用人体 EVLP 模型重复这些实验。对肺组织进行全面的表型、转录和功能终点评估以及 BRx 中的肝细胞，例如 RAGE 和 NF-κB 如果成功，我们的技术将提供意味着改变EVLP的当前状态以进行肺保存并让肺得以维持最终，我们的工作将解决当前的问题。 LTx 的局限性，包括增加可行的供体器官运输时间和距离，帮助减少 LTx 等待名单，并提高移植后患者的生存率。