LIVER REPOPULATION FOR LIVER INJURY AND GENE THERAPY

肝损伤的肝脏再生和基因治疗

• 批准号: 2146261
• 负责人: SANJEEV GUPTA
• 金额: $ 24万
• 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/2146261
• 关键词: autocrine; cell growth regulation; embryonic stem cell; gene therapy; genetic models; growth factor; hemodynamics; 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; autocrine; cell growth regulation; embryonic stem cell; gene therapy; genetic models; growth factor; hemodynamics; 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.

肝细胞移植具有基因治疗的巨大潜力和 用于治疗急性肝衰竭。因为肝细胞具有细胞 表达各种基因的机械，它们用于前体基因 治疗特别有吸引力。同样，肝细胞移植 应该为急性肝衰竭提供新颖的疗法，目前 导致不可接受的死亡率大于80％。虽然 原位肝移植可用于急性肝衰竭， 这是一个强大而不可逆转的程序。相反，优势 肝细胞的移植包括其技术简单性， 从单个肝脏制备的移植细胞的可能性 多个接受者以及冷冻和移植细胞的潜力 在短时间内。但是，尽管施加肝细胞非常紧迫 人类的移植，对生物学的更好理解 最佳用法需要移植的肝细胞。 我们假设肝细胞移植的成功需要a） 识别肝细胞生存的允许条件和 功能，b）识别增加质量的方法 移植的肝细胞，c）优化时间和剂量 治疗应用所需的细胞。开始解决这些 关键问题，我们开发了具有遗传标记的新型系统 肝细胞明确区分移植和宿主 细胞。此外，我们证明了移植的肝细胞生存，并且 在肝脏中功能最佳。我们现在建议检查机制 调节肝脏移植的肝细胞的增殖和命运。我们 将检查移植的肝细胞的增殖能力和 确定使用肝脏再生刺激是否可以增加质量 移植的肝细胞。作为将肝细胞移植到 肝脏可能受到肝血管床的容量的限制，我们将 确定肝细胞移植的安全性。使用得出的见解 从这些研究中，我们将制定大规模重组的策略 宿主肝。这些策略将在强大的动物中进行测试 遗传代谢疾病的模型。另外，急性肝动物模型 失败将用于定义肝细胞的时机，剂量和价值 移植，同时肝增长 因素。最后，将使用肝细胞前体细胞来确定 它们的价值与原发性肝细胞相比。这些完成 拟议的研究将大大提高我们对 肝细胞移植并有助于将这项技术从 床边的实验室。