FAH-Deficient Pigs

FAH 缺陷猪

基本信息

批准号：
8250747
负责人：
SCOTT L NYBERG
金额：
$ 21.24万
依托单位：
YECURIS CORPORATION
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-20 至 2013-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8250747
关键词：
Accounting Acute Liver Failure Address Adverse effects Affect American Animal Model Annual Reports Area Artificial Liver Basic Science Biomedical Engineering Bioreactors Biotechnology Breeding Cause of Death Cell Transplantation Cells Cessation of life Chronic Cirrhosis Clinical Trials Code Development Devices Diagnostic Disease Dose Drug usage Engraftment Excision Family suidae Female Fibrosis Foundations Fumarylacetoacetase Future Gene Targeting Genes Goals Health Hepatitis Hepatocyte Heterozygote Homozygote Human In Vitro Inborn Errors of Metabolism Injection of therapeutic agent Injury Knock-out Liver Liver Cirrhosis Liver Failure Liver diseases Malignant Neoplasms Metabolic Metabolic Diseases Methodology Modality Modeling Mus Natural regeneration Nature Nodule Organ Donor Pharmaceutical Preparations Pharmacologic Substance Phase Phenotype Primary carcinoma of the liver cells Process Production Protective Agents Recombinant adeno-associated virus (rAAV)Refractory Research Research Design Research Personnel Resource Development Rodent Model Safety Screening procedure Serotyping Signal Transduction Small Business Technology Transfer Research Source System Testing Therapeutic Toxic effect Toxicity Tests Transgenic Organisms Translational Research Transplantation Tyrosine Metabolism Pathway Tyrosinemias United States base catalyst commercialization drug discovery effective therapy human disease immunoregulation in vivo large scale production liver cell proliferation liver transplantation metabolic abnormality assessment mutant new technology novel novel strategies novel therapeutics phase 1 study phase 2 study pressure prevent programs prototype regenerative research and development scale up somatic cell nuclear transfer tumor

项目摘要

DESCRIPTION (provided by applicant): Translational research in the field of liver disease has been limited by a shortage of large animal models, which in turn has limited development of new therapies for metabolic liver disorders, acute liver failure, chronic (cirrhotic) liver disease, an hepatocellular carcinoma. While new transgenic and knockout rodent models of liver disease have facilitated basic research, the development of bioengineered large animal models of liver disease has been problematic. To address the shortage of large animal models, we have utilized a novel strategy combining gene targeting by recombinant adeno-associated virus DJ serotype (rAAVdj) and somatic cell nuclear transfer (SCNT) for the production of bioengineered pigs. Our prototype model is hereditary tyrosinemia type 1 (HT1) accomplished by knocking out the gene coding for fumarylacetoacetate hydrolase (FAH) in pigs. In humans, FAH deficiency is associated with a unique phenotype, which includes metabolic derangement (tyrosinemia), acute liver failure, cirrhosis, and hepatocellular carcinoma (HCC). In mice, FAH deficiency combined with immune modulation has been used to produce mice with humanized livers (Azuma 2007 Nature Biotechnology; Bissig 2010 J Clinical Investigation). These FAH deficient mice serve as in vivo bioreactors for the robust expansion of normal human hepatocytes. We postulate that scale up of the FAH deficient platform to pigs is possible, and that FAH deficient pigs will also serve as surrogate hosts for much larger scale in vivo expansion of normal human hepatocytes. An abundant supply of human hepatocytes is needed for applications including toxicity testing of new drugs, human hepatocyte transplantation, and cell- based liver support devices such as the bioartificial liver. With regard to commercialization of this novel technology, we have already produced Fah-null heterozygote female pigs by rAAVdj and SCNT methodology. This phase 1 STTR (R41) application will establish a herd of FAH-deficient pigs and characterize the phenotype of Fah-null homozygote pigs. In future phase 2 studies, FAH-deficient pigs will be evaluated as in vivo bioreactors for the large-scale production of human hepatocytes. We expect that the FAH-deficient pig will serve as a valuable resource for the development of novel therapeutic modalities and to facilitate translational research in the field o liver disease. Furthermore, the rAAVdj and SCNT methodology may be utilized to produce other bioengineered large animal models of human disease. PUBLIC HEALTH RELEVANCE: The goal of the current research program is to develop a bioengineered pig with the metabolic disorder of human hereditary tyrosinemia type 1 (HT1). Phase 1 studies are designed to establish a herd of HT1 pigs and characterize the phenotype of these pigs. Phase 2 studies will determine if HT1 pigs can serve as in vivo bioreactors for large scale production of human hepatocytes. There are many therapeutic applications for normal human hepatocytes such as a cell-source in a bioartificial liver or for hepatocyte transplantation in humans, or industrial applications for safety and toxicity screening, or for use by pharmaceutical companies in the drug discovery process. Thus the demand for these new pigs is worthy of commercialization.

描述（由申请人提供）：肝病领域的转化研究受到大型动物模型的短缺的限制，而大型动物模型又有有限的代谢性肝疾病，急性肝衰竭，慢性（肝硬化）肝病，肝细胞癌癌的发展有限。尽管肝病的新转基因和敲除啮齿动物模型促进了基础研究，但生物工程的大型肝病动物模型的发展一直是有问题的。为了解决大型动物模型的短缺，我们利用了一种新的策略，通过重组腺相关病毒DJ血清型（RAAVDJ）和体细胞核转移（SCNT）结合了基因靶向，以生产生物工程猪。我们的原型模型是通过敲除编码猪富马乙酸乙酸酯水解酶（FAH）的基因来完成的遗传性酪氨酸1型（HT1）。在人类中，FAH缺乏症与独特的表型有关，其中包括代谢危险（酪蛋白血症），急性肝衰竭，肝硬化和肝细胞癌（HCC）。在小鼠中，FAH缺乏症与免疫调节相结合已用于用人源化肝脏产生小鼠（Azuma 2007自然生物技术； Bissig 2010 J临床研究）。这些FAH缺乏的小鼠是正常人肝细胞稳健膨胀的体内生物反应器。我们假设可以扩大到FAH缺乏猪的平台对猪的规模，而FAH缺乏的猪也将作为正常人肝细胞体内扩张较大的体内扩张量更大的替代宿主。对于应用，包括新药的毒性测试，人肝细胞移植和基于细胞的肝脏支持装置（例如生物人工肝脏）的应用，需要大量的人肝细胞供应。关于这种新技术的商业化，我们已经通过Raavdj和Scnt方法生产了Fah-null杂合雌性猪。该阶段1 STTR（R41）的应用将建立一群缺陷的猪，并表征Fah-null纯合猪的表型。在未来的第二阶段研究中，缺乏FAH的猪将作为体内生物反应器评估，用于大规模生产人类肝细胞。我们预计，缺乏FAH的猪将成为开发新型治疗方式的宝贵资源，并促进肝病野外转化研究。此外，RAAVDJ和SCNT方法可以用于产生其他生物工程的大型人类疾病动物模型。公共卫生相关性：当前研究计划的目的是开发一种生物工程的猪，该猪具有人类遗传性酪氨酸1型（HT1）的代谢疾病。第1阶段的研究旨在建立HT1猪的群，并表征这些猪的表型。第2阶段的研究将确定HT1猪是否可以用作人体肝细胞大规模生产的体内生物反应器。对于正常的人工肝中的细胞来源或人类的肝细胞移植，有许多治疗应用，例如人工肝中的细胞来源，或用于安全性和毒性筛查的工业应用，或用于药物发现过程中的制药公司使用。因此，对这些新猪的需求值得商业化。