Immunologic Biomarkers of Rejection in Clinical Liver Transplantation

项目摘要 候选人：Juliet Emmaullee是南方大学临床外科助理教授 加州（南加州大学）和凯克医院和儿童医院的一名主治肝肾移植外科医生- 洛杉矶。Emmaullee博士一直在与她提议的K 08导师Omid Akbari博士和她的合作伙伴一起工作。 导师Shahab Asgharzadeh博士，了解用于表征免疫反应的质谱细胞术方法 现在已经开始独立工作，开发这些技术来分析肝脏中的排斥反应过程 移植（LT）受者。该项目旨在为她提供额外的技能和知识， 实现她的长期目标，研究参与发展和进展的免疫机制， 为了开发新的诊断，预防和治疗方法，以改善LT后 结果。 职业发展计划：Emmaullee博士已制定战略计划，以获得必要的培训， 她成功过渡到独立调查员所需的指导， 课程和强大的指导计划。她还组织了一个咨询委员会， 领导人在该领域，但也有那些能够直接影响她的职业发展。立即培训 目标是集中在巩固她的专业知识：（1）先进的免疫技术，包括质量 细胞计数;（2）生物信息学和数据分析;以及（3）临床LT中排斥反应的免疫学途径。 这不仅将确保Emmaullee博士的研究项目按计划进行，而且还将确保她 将通过推广和获得独立研究资金来确认取得的进展。她有一个 有影响力的，独特的研究项目，与导师的研究有足够的不同，以避免竞争， 重叠 研究计划：拟议的研究利用南加州大学的广泛资源来解决一个重要的 公共卫生问题，这使得它直接关系到国家卫生研究院的使命。肝细胞癌的发病率 成为LT的主要全球适应症，排斥反应仍然是移植物丢失的重要原因， 移植后失败排斥反应的诊断与肝脏血液检查的变化无关， 需要进行侵入性活检最近，在技术的敏感性方面有了重大进展， 表征免疫反应，即使是在小的组织样品中。我们假设肝脏免疫系统 可以使用成像质谱细胞术（IMC）开发面板，以深入分析肝内免疫浸润 在人类肝脏组织中。我们还假设移植物浸润淋巴细胞可以用IMC来表征， 更好地了解哪些亚群介导LT中的同种异体移植排斥反应。首先，我们将建立一个肝脏 免疫学IMC面板使用从具有慢性排斥的LT患者获得的组织。此外，我们将利用 本试验旨在描述慢性排斥反应中移植物浸润淋巴细胞亚群的特征，并与 使用从具有急性排斥反应的LT患者获得的组织获得。排斥反应各阶段的免疫特征 将被量化和映射，以及与临床参数进行比较。最后，我们将研究LT患者 前瞻性地确定这些排斥相关淋巴细胞是否可以在血液中追踪， 拒绝的情节。该项目代表了IMC在实体器官移植中的首次应用， 扩大我们对临床LT中同种免疫的认识的潜力，这可以为临床LT提供基础。 在LT患者中的排斥反应的非侵入性测定。

PROJECT SUMMARY Candidate: Juliet Emamaullee is an Assistant Professor of Clinical Surgery at the University of Southern California (USC) and an attending liver and kidney transplant surgeon at Keck Hospital and Children’s Hospital- Los Angeles. Dr. Emamaullee has been working with her proposed K08 mentor, Dr. Omid Akbari, and her co- mentor, Dr. Shahab Asgharzadeh, to learn about mass cytometry approaches to characterize immune responses and now has begun independent work to develop these techniques to analyze the process of rejection in liver transplant (LT) recipients. The project aims to provide her with additional skills and knowledge required to achieve her long-term goal of studying the immunologic mechanisms involved in development and progression of LT rejection in order to develop new diagnostic, preventative, and therapeutic approaches to improve post-LT outcomes. Career Development Plan: Dr. Emamaullee has strategically planned to gain the necessary training and mentoring that will be required for her successful transition to being an independent investigator through select coursework and a robust mentoring plan. She has also organized an advisory committee composed not only of leaders in the field but also those able to directly impact her career advancement. The immediate training objectives are focused on consolidating her expertise in: (1) advanced immunological techniques including mass cytometry; (2) bioinformatics and data analysis; and (3) immunological pathways of rejection in clinical LT. This will not only ensure that Dr. Emamaullee's research project progresses as planned but will also ensure her progress will be recognized through promotion and obtaining independent research funding. She has an impactful, unique research project that is sufficiently different from her mentor's research to avoid competition or overlap. Research Plan: The proposed study leverages the extensive resources available at USC to address an important public health issue, which makes it directly relevant to the NIH mission. Hepatocellular carcinoma is on pace to become the leading global indication for LT, and rejection continues to be an important cause of graft loss and failure post-transplant. The diagnosis of rejection is not correlative to changes in liver blood tests and thus requires an invasive biopsy. Recently, there have been significant advances in the sensitivity of techniques to characterize immune responses, even in small tissue samples. We hypothesize that a liver-focused immune panel can be developed using imaging mass cytometry (IMC) to deeply analyze intrahepatic immune infiltrates in human liver tissue. We also hypothesize that graft-infiltrating lymphocytes can be characterized using IMC to better understand which subpopulations mediate allograft rejection in LT. First, we will develop a Liver Immunology IMC Panel using tissue obtained from LT patients with chronic rejection. Furthermore, we will utilize this assay to characterize graft-infiltrating lymphocyte subpopulations in chronic rejection and compare to results obtain using tissue obtained from LT patients with acute rejection. Immune profiles in each phase of rejection will be quantified and mapped, as well as compared to clinical parameters. Lastly, we will study LT patients prospectively to determine if these rejection-associated lymphocytes can be tracked in the blood during and after episodes of rejection. This project represents the first application of IMC in solid organ transplantation and has the potential to expand our knowledge of alloimmunity in clinical LT, which could provide the basis for a noninvasive assay of rejection in LT patients.