Use of Discarded Organs for Preparation of Liver Grafts

使用废弃器官制备肝移植物

• 批准号: 8301550
• 负责人: Basak Elif Uygun
• 金额: $ 8.74万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8301550
• 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; Solutions; Source; Spatial Distribution; Staining method; Stains; Stem cells; Structure of beta Cell of islet; 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; tool

DESCRIPTION (provided by applicant): 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 beta-cell transplantation), and may ultimately lead to development of entire organs in vitro.

描述（由申请人提供）：终阶段器官故障的治疗受到当前以100,000请求的器官候补名单的严重短缺的限制，并且每年增加5％。对于肝脏而言，该研究的重点是没有什么不同 - 由于缺乏移植器官，在美国大约有4,000人死亡，缺乏供体器官被认为是重大的健康危机。另外，由于移植通常可以解决许多与衰老有关的疾病的解决方案，因此估计隐藏的需求远远超出了当前水平。这种情况一直是组织工程兴起的主要推动力，其器官衰竭市场估计约为800亿美元。但是，旨在从头开始建立组织的二十年的工作并没有成功地创建可以在临床上植入的大规模组织，以解决器官替代疗法中的空隙。此外，尽管在包括我们小组的干细胞方面进行了巨大的努力，但缺乏用于细胞疗法的原发性成人肝细胞的可靠细胞来源持续存在，并且在不久的将来不太可能解决。有趣的是，由于器官损坏，有许多潜在的器官捐献者不被考虑进行移植。例如，由于缺血性损害过多，心脏死亡后到达医院的事故受害者没有资格捐助者。众所周知，即使是轻微的缺血损伤（> 30分钟温暖的缺血或> 16小时）也会导致并发症，并在6个月时大大降低移植物存活率。据某些估计，在美国，心脏死亡后的潜在捐助者人数超过200,000，而大约6,000人被认为仅受到少量损害。我们的长期目标是设计用于治疗或治疗相关肝病的可移植肝移植物。拟议的研究的目的是开发人性化重新设计的肝移植物作为可行的体外肝模型。在该奖项的指导阶段（K99）中，将开发两个基本工具以实现这一目标：1）肝脏灌注系统，这将使从边缘供体肝脏中恢复健康的肝细胞。预计这项技术将导致建立目前尚未开发的成年人类肝细胞来源，该来源将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使用。 2）整个器官灌注 - 脱细胞和卷积方法。这里的目的是为组织工程开发一种新型的脚手架，该脚手架支持细胞的附着和功能，并且是可血管化的。在该奖项的独立阶段（R00）期间，主要目标是将K99阶段中开发的方法扩展到大型动物模型和最终人体器官。 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.虽然这项工作利用肝脏作为模型器官，但这项工作的结果还将通过建立未来复杂的器官工程技术的基础来产生积极的影响，这些器官工程技术结合了多种不同的细胞类型，并可以转化为其他器官（例如胰腺（例如为胰腺beta beta-beta-cell cell创建血管化斑块），以最终导致整个Organs的开发。