Use of Discarded Organs for Preparation of Liver Grafts

使用废弃器官制备肝移植物

基本信息

批准号：
8974403
负责人：
Basak Elif Uygun
金额：
$ 24.9万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-12-17 至 2017-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8974403
关键词：
Accidents Address Adult Affect Albumins Animal Model Architecture Award Biocompatible Materials Biological Preservation Biological Process Biology Cardiac Death Cell Culture Techniques Cell Therapy Cell Transplantation Cell physiology Cell-Matrix Junction Cells Clinical Cytochrome P450 Detergents Development Disease Drug Metabolic Detoxication Engineering Extracellular Matrix Family suidae Future Glycoproteins Goals Graft Survival Health Hepatic Hepatic Tissue Hepatocyte Hospitals Human Image Analysis Implant In Vitro Injury Ionic Strengths Ischemia Lead Liver Liver diseases Marketing Mentors Methodology Methods Modality Modeling New England Organ Organ Donor Organ Model Organ Transplantation Organ failure PECAM1 gene Pancreas Perfusion Pharmacologic Substance Phase Population Preparation Proteoglycan Protocols documentation Rattus Recovery Replacement Therapy Research Role Slice Source Spatial Distribution Staining method Stains Stem cells Supporting Cell System Techniques Technology Testing Tissue Engineering Tissues Translating Transplantation Urea Waiting Lists Warm Ischemia Work age related base cell type driving force drug testing end-stage organ failure in vitro Model in vivo innovation liver transplantation novel scaffold scale up three dimensional cell culture tool

项目摘要

PROJECT SUMMARY/ABSTRACT Treatment for end-stage organ failure is restricted by the critical shortage of donor organs with the organ waiting list currently at 100,000 requests and it is increasing by 5% every year. The problem is not different for liver, which this study focuses on - about 4,000 people die in the US due to lack of a transplantable organ, and the lack of donor organs is considered a major health crisis. In addition, since transplantation can often be the solution to many aging related diseases, the hidden demand is estimated to be far beyond the current levels. This situation has been a major driving force behind the rise of tissue engineering, with the market for organ failure treatments estimated at about $80 billion. However, over two decades of work aimed at building tissues from the ground up has not succeeded in creating large-scale tissues that can be clinically implanted to address the void in organ replacement therapies. Further, despite intense efforts on the stem cell front, including those from our group, the lack of a reliable cell source for primary adult hepatocytes for use in cell therapies persists and is unlikely to be resolved in the near future. Interestingly enough, there are many potential organ donors that are not considered for transplantation because the organs are damaged. For example, accident victims who arrive at the hospital after cardiac death are not eligible donors because of excessive ischemic damage; even a slight ischemic damage (>30min warm ischemia or >16hrs cold ischemia) is known to lead to complications in the long term with significantly reduced graft survival at 6 months. By some estimates, potential number of Donors after Cardiac Death (DCD) is over 200,000 per year in the US, and about 6,000 are considered to be only marginally damaged. Our long-term goal is to engineer transplantable liver grafts for curing or treating relevant liver diseases. The objective of the proposed study is to develop humanized reengineered liver grafts as a viable in vitro liver model. During the mentored phase (K99) of the award, two essential tools will be developed to reach this goal: 1) a liver perfusion system, which will enable recovery of healthy hepatocytes from marginal donor livers. This technology is expected to lead to establishment of a currently untapped source of adult human hepatocytes that will fill the need for human cells until stem cell approaches mature and become safe and efficient for clinical use. 2) a whole organ perfusion- decellularization and recellularization methodology. The objective here is to develop a novel scaffold for tissue engineering, which supports cell attachment and function and is vascularizable. During the independent phase (R00) of the award, the primary goal is the scaling of the methods developed in K99 phase to large animal models and ultimately human organs. The work proposed in this project is expected to i) establish marginal livers as a reliable source of primary hepatocytes, ii) establish decellularized liver slices as novel 3D cell culture platform to study the role of ECM, iii) develop humanized rat liver grafts as a three dimensional liver model for pharmaceutical studies, and iv) lead to the development of reengineered liver grafts to treat liver diseases. While this work utilizes liver as the model organ, the results of this work will also have a positive impact by establishing the basis of future sophisticated organ engineering techniques that incorporate multiple different cell types and can be translated to other organs (such as pancreas to create vascularized patches for pancreatic ¿-cell transplantation), and may ultimately lead to development of entire organs in vitro.

项目概要/摘要终末期器官衰竭的治疗受到供体器官严重短缺的限制目前列表中有 100,000 个请求，并且每年以 5% 的速度增长等待的问题也没有什么不同。肝脏，本研究的重点 - 在美国约有 4,000 人因缺乏可移植器官而死亡，以及此外，缺乏捐赠器官被认为是一个重大的健康危机。对于许多与衰老相关的疾病的解决方案，隐性需求估计将远远超出目前的水平。这种情况一直是组织工程兴起的主要推动力，器官市场失败治疗估计耗资约 800 亿美元然而，二十多年来致力于构建组织的工作。从头开始尚未成功地制造出可以临床植入的大规模组织此外，尽管在干细胞方面付出了巨大努力，但仍解决了器官替代疗法的空白。包括我们组的人员，缺乏可靠的原代成体肝细胞来源以用于细胞治疗持续存在，并且不太可能在不久的将来得到解决。由于器官受损而不考虑移植的潜在器官捐献者。例如，心脏死亡后到达医院的事故受害者不符合捐赠者的资格，因为过度缺血性损伤；甚至轻微缺血性损伤（>30分钟热缺血或>16小时冷缺血）众所周知，从长远来看，这会导致并发症，导致移植物的 6 个月存活率显着降低。据估计，美国每年心脏死亡 (DCD) 后的潜在捐赠者数量超过 200,000 人，并且大约 6,000 只被认为仅受到轻微损坏，我们的长期目标是设计可移植的。本研究的目的是开发用于治愈或治疗相关肝脏疾病的肝移植物。在指导阶段（K99），人性化重组肝移植物作为可行的体外肝脏模型。奖项，将开发两个基本工具来实现这一目标：1）肝脏灌注系统，这将使该技术有望从边缘供体肝脏中恢复健康肝细胞。建立目前尚未开发的成人肝细胞来源，以满足人类细胞的需求直到干细胞接近成熟并变得安全有效地用于临床2）全器官灌注-。脱细胞和再细胞化方法的目的是开发一种新型的组织支架。工程，支持细胞附着和功能，并且在独立阶段具有血管化能力。 (R00) 的奖项，主要目标是将 K99 阶段开发的方法扩展到大型动物模型和最终的人体器官预计该项目中提出的工作 i) 建立边际。肝脏作为原代肝细胞的可靠来源，ii) 建立脱细胞肝脏切片作为新型 3D 细胞培养平台来研究 ECM 的作用，iii) 开发人源化大鼠肝移植物作为三维肝脏药物研究模型，以及 iv) 导致开发用于治疗肝脏的再造肝移植物虽然这项工作利用肝脏作为模型器官，但这项工作的结果也将具有积极的意义。通过建立未来复杂的器官工程技术的基础来影响，这些技术融合了多种不同的细胞类型，并且可以转化为其他器官（例如胰腺，以创建血管化斑块胰-细胞移植），并可能最终导致整个器官在体外发育。