INNOVATIVE APPROACHES TO THE STUDY OF LIVER REGENERATION

说明(改编自调查员的摘要)本提案寻求 进一步研究外科生物学的一个基本问题，那就是肝脏 利用创新技术研究再生机制 牵涉其中。该提案的陈述假设包括：1)重组 腺病毒是最有效和最广泛适用的肝靶向病毒 基因转移载体；2)DNA结合蛋白，特别是NK-B和STAT-3 在肝脏再生中发挥了重要作用，而且很可能还 重组腺病毒基因在肝脏中的表达；3)延长的基因 再生肝中的表达不是由于免疫功能受损 反应；4)基因转移可以用来引入具有重要生物学意义的 基因，特别是角质形成细胞生长因子和肝细胞生长因子， 在体外进入肝细胞，在体内进入正常肝脏，允许 阐明肝脏增殖的机制；5)基因 移位可用于研究心脏的生理学和病理生理学。 肝脏的再生和葡萄糖代谢。这样做的具体目的是 建议包括：1)分析载体效率、基因的模式 正常、再生和碳水化合物的表达和宿主免疫反应 用含四氯化碳的重组腺病毒处理小鼠肝脏 标记基因；2)研究核结合蛋白的表达。 分离的小鼠肝细胞，正常肝脏，重组基因转导的肝脏 腺病毒，与再生肝；3)研究 延长的重组细胞中的抗细胞凋亡因子Bcl2和Bclxl 再生肝中可见的腺病毒介导的基因转移；4)分析 转基因(-半乳糖苷酶)标记的肝细胞对 重组蛋白存在或不存在时的小鼠肝脏移植 表达角质形成细胞生长因子和/或肝细胞生长的腺病毒 因子；5)研究肝脏导向的重组腺病毒的作用 葡萄糖激酶介导性过表达对葡萄糖稳态和肝脏的影响 链脲佐菌素诱导糖尿病小鼠假手术后或70%后的再生 肝切除手术。

DESCRIPTION (Adapted from the investigator's abstract) This proposal seeks to further study a fundamental problem of surgical biology, that is, liver regeneration using innovative techniques for the study of the mechanisms involved. The stated hypotheses of this proposal include: 1) recombinant adenoviruses are the most efficient and broadly-applicable liver-directed gene transfer vectors; 2) DNA binding proteins, especially NK-(B and STAT-3 have assumed an important role in liver regeneration and are likely to also be important in adenoviral gene expression in liver; 3) the prolonged gene expression seen in regenerating liver is not due to an impaired immune response; 4) gene transfer can be used to introduce biologically important genes, specifically keratinocyte growth factor and hepatocyte growth factor, into hepatocytes in vitro, and normal liver in vivo, allowing the elucidation of mechanisms responsible for liver proliferation; and 5) gene transfer can be used to study the physiology and pathophysiology of regeneration and glucose metabolism in the liver. The Specific Aims of this proposal include: 1) to analyze the patterns of vector efficiency, gene expression and host immune response in normal, regenerating, and carbon tetrachloride-treated mouse liver using recombinant adenoviruses containing marker genes; 2) to study the expression of nuclear binding proteins in isolated mouse hepatocytes, normal liver, liver transduced with recombinant adenovirus, and regenerating liver; 3) to study the role of the anti-apoptosis factors Bcl-2, and Bcl-XL, in the prolonged recombinant adenovirus-mediated gene transfer seen in regenerating liver; 4) to analyze the ability of hepatocytes marked with the (-galactosidase transgene to engraft in mouse liver in the presence or absence of recombinant adenoviruses expressing keratinocyte growth factor and/or hepatocyte growth factor; and 5) to study the effects of liver-directed recombinant adenovirus mediated overexpression of glucokinase on glucose homeostasis and hepatic regeneration in mice made diabetic by streptozotocin after sham or 70% hepatectomy.