Targeting Donor Regulatory Dendritic Cells During Normothermic Ex Vivo Liver Perfusion to Overcome Rejection after Liver Transplant

摘要 该提案提出了一项为期五年的研究职业发展计划，重点是针对供体肝脏- 具有调节特性的驻留细胞以减少移植后的排斥。我是助理教授 威斯康星大学麦迪逊分校外科博士，具有移植方面的研究和临床经验 涉及常温离体机器灌注（NEVLP）的免疫学和移植手术， 器官被置于生理条件下。本项目将推动移植领域的发展 免疫学通过使用NEVLP技术在移植前修饰肝脏内的免疫细胞。我 组建了一支由具有移植免疫学专业知识的研究人员组成的优秀导师团队， 树突细胞生物学和细胞外囊泡生物学。拟议的培训将指导和提高我的 核心能力的发展，包括移植免疫学，通信，生物统计学和伦理 研究设计，这将使我能够过渡到研究独立作为一个外科医生，科学家致力于 减少移植手术领域的器官排斥。 肝移植是终末期肝病患者的唯一治疗选择;然而， 移植排斥会降低肝脏和患者的存活率。此外，患者仍要求终身使用 抑制免疫系统的抗排斥药物。对供体肝脏进行改造， 细胞，有可能促进肝脏的接受，并最大限度地减少抗排斥反应的需要。 毒品一种称为常温离体肝脏灌注（NEVLP）的创新技术的进展提供了一种独特的 有机会使25%发生急性排斥反应的肝移植受者显著受益， 更多的移植受者将受益于使用更少的抗排斥药物。最近的研究 证实了调节性树突状细胞（DCDC）对于延长移植物存活的重要性。我 中心假设是NEVLP期间肝内DCIII数量的增加将促进 器官移植后的监管环境。使用NEVLP和肝移植的大鼠模型， 我的研究小组已经优化了，我预计NEVLP将有力地扩大DCCP，从而增加 免疫检查点分子表达和抗炎细胞外囊泡的产生， 可以减少免疫介导的排斥反应的细胞因子。这种创新的方法扩大移植驻留 减少排斥反应的DCCl可用于死亡供体肝移植，也可转化为 其他实体器官移植。为了实现这些目标，我提出以下科学目标： 1)确定NEVLP后肝内DCCl的主要调节功能，2）测量 NEVLP期间联合细胞因子治疗产生的扩展的肝驻留DC-DC对肝脏的影响 在体外和体内的移植排斥。

ABSTRACT This proposal presents a five-year research career development program focused on targeting donor liver- resident cells with regulatory properties to decrease rejection after transplantation. I am an Assistant Professor of Surgery at the University of Wisconsin-Madison, with previous research and clinical experience in transplant immunology and transplant surgery involving normothermic ex vivo machine perfusion (NEVLP), whereby an organ is housed under physiologic conditions. The present project will advance the field of transplant immunology by using NEVLP technology to modify the immune cells within the liver prior to transplantation. I have assembled an outstanding mentorship team of investigators with expertise in transplant immunology, dendritic cell biology, and extracellular vesicle biology. The proposed training will guide and enhance my development in core competencies, including transplant immunology, communication, biostatistics, and ethical research design that will enable me to transition to research independence as a surgeon-scientist dedicated to reducing organ rejection in the field of transplant surgery. Liver transplantation is the only treatment option for patients with end-stage liver disease; however, rejection of the transplant can decrease liver and patient survival. In addition, patients still require lifelong use of anti-rejection medications that suppress the immune system. Modification of the donor liver, and the immune cells within it, has the potential to promote acceptance of the liver and minimize the need for anti-rejection drugs. Advances in an innovative technique called normothermic ex vivo liver perfusion (NEVLP) offer a unique opportunity to benefit significantly the 25% of liver transplant recipients that develop acute rejection, as well as many more transplant recipients who would benefit from using fewer anti-rejection drugs. Recent studies have demonstrated the importance of regulatory dendritic cells (DCregs) for prolonging transplant survival. My central hypothesis is that expansion of the number of liver-resident DCregs during NEVLP will promote a regulatory environment for the organ after transplant. Using a rat model of NEVLP and liver transplantation that my research group has optimized, I expect NEVLP to expand DCregs potently, leading to an increase in immune checkpoint molecule expression and production of anti-inflammatory extracellular vesicles and cytokines that can reduce immune-mediated rejection. This innovative approach of expanding graft-resident DCregs to decrease rejection could be used in deceased donor liver transplantation as well as translated to other types of solid organ transplants. To achieve these objectives, I propose the following scientific aims: 1) Determine the dominant regulatory function of liver-resident DCregs after NEVLP, and 2) Measure the impact of expanded liver-resident DCregs generated by combination cytokine therapy during NEVLP on liver graft rejection in vitro and in vivo.