LIVER REPOPULATION FOR LIVER INJURY AND GENE THERAPY

肝损伤的肝脏再生和基因治疗

• 批准号: 2414851
• 负责人: SANJEEV GUPTA
• 金额: $ 30.53万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-06-20 至 1998-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2414851
• 关键词: autocrine; cell growth regulation; embryonic stem cell; gene therapy; genetic models; hemodynamics; hepatocyte growth factor; inborn metal metabolism disorder; injury; laboratory mouse; laboratory rat; liver cells; liver failure; liver regeneration; liver transplantation; nonhuman therapy evaluation; stimulus /response; tissue /cell culture; transforming growth factors

Hepatocyte transplantation has an immense potential for gene therapy and for treating acute liver failure. Because hepatocytes possess the cellular machinery to express a variety of genes, their use for ex-vivo gene therapy is particularly attractive. Similarly, hepatocyte transplantation should provide novel therapies for acute liver failure, which currently results in unacceptable mortalities of greater than 80%. Although orthotopic liver transplantation can be employed in acute liver failure, this is a formidable and irreversible procedure. In contrast, advantages of hepatocyte transplantation include its technical simplicity, the possibility of transplanting cells prepared from a single liver into multiple recipients and the potential to cryopreserve and transplant cells at a short notice. However, despite a great urgency in applying hepatocyte transplantation in humans, a better understanding of the biology of transplanted hepatocytes is required for an optimal usage. We hypothesize that the success of hepatocyte transplantation requires a) identification of permissive conditions for hepatocyte survival and function, b) identification of methods to increase the mass of transplanted hepatocytes, and c) optimization of the time and dose of cells required for therapeutic applications. To begin addressing these critical issues, we developed novel systems with genetically marked hepatocytes to unequivocally distinguish between transplanted and host cells. Also, we demonstrated that transplanted hepatocytes survive and function most optimally in the liver. We now propose to examine mechanisms regulating proliferation and fate of transplanted hepatocytes in liver. We will examine the proliferative capacity of transplanted hepatocytes and determine whether use of liver regenerative stimuli could augment the mass of transplanted hepatocytes. As transplantation of hepatocytes into the liver may be limited by the capacity of the hepatic vascular bed, we will determine the safety of hepatocyte transplantation. Using insights derived from these studies, we will develop strategies for massive reconstitution of the host liver. These strategies will be tested in powerful animal models of genetic metabolic disease. Also, animal models of acute liver failure will be used to define the timing, dose and value of hepatocyte transplantation, a well as simultaneous administration of hepatic growth factors. Finally, hepatocyte precursor cells will be used to determine their value compared with primary hepatocytes. Completion of these proposed studies will greatly advance our fundamental knowledge of hepatocyte transplantation and help move this technology from the laboratory to the bedside.

肝细胞移植在基因治疗和治疗方面具有巨大的潜力 用于治疗急性肝功能衰竭。因为肝细胞拥有细胞 机械表达多种基因，它们用于体外基因 治疗尤其吸引人。同样，肝细胞移植 应该为急性肝功能衰竭提供新的治疗方法，目前 导致超过80%的不可接受的死亡率。虽然 原位肝移植可用于急性肝功能衰竭， 这是一个艰巨且不可逆转的过程。相比之下，优势 肝细胞移植的技术简单性， 将从单个肝脏制备的细胞移植到 多个受体和冷冻保存和移植细胞的可能性 在很短的时间内。然而，尽管应用肝细胞迫在眉睫 在人类身上进行移植，更好地理解了 最佳使用需要移植的肝细胞。 我们假设肝细胞移植的成功需要一个) 肝细胞存活的允许条件的确定和 功能，b)确定增加质量的方法 移植肝细胞，以及c)时间和剂量的优化。 治疗应用所需的细胞。开始解决这些问题 关键问题，我们开发了带有基因标记的新型系统 肝细胞明确区分移植和宿主 细胞。此外，我们还证明了移植的肝细胞存活并 在肝脏中发挥最好的功能。我们现在建议研究这些机制 调节肝内移植肝细胞的增殖和命运。我们 将检测移植的肝细胞的增殖能力 确定使用肝再生刺激是否可以增加质量 移植的肝细胞。作为肝细胞移植到 肝脏可能会受到肝脏血管床容量的限制，我们将 确定肝细胞移植的安全性。利用得出的见解 从这些研究中，我们将制定大规模重建的策略 寄主肝脏的。这些策略将在强大的动物身上进行测试 遗传代谢性疾病的模型。此外，急性肝病的动物模型 失败将被用来定义肝细胞移植的时机、剂量和价值 移植，一种同时管理肝脏生长的好方法 各种因素。最后，将使用肝细胞前体细胞来确定 与原代肝细胞相比，其价值更高。完成这些工作 拟议的研究将极大地提高我们对 肝细胞移植并帮助将这项技术从 从实验室到床边。