INNOVATIVE APPROACHES TO THE STUDY OF LIVER REGENERATION

描述（改编自研究者摘要） 为了进一步研究外科生物学的一个基本问题，即肝脏， 再生利用创新技术的机制的研究 涉案 该提案的假设包括：1）重组 腺病毒是最有效和最广泛适用的肝定向 基因转移载体; 2）DNA结合蛋白，特别是NK-（B和STAT-3 在肝再生中起着重要作用， 在腺病毒基因在肝脏中的表达中起重要作用; 3）延长的基因 在再生肝脏中观察到的表达不是由于免疫受损 反应; 4）基因转移可用于引入生物学上重要的 基因，特别是角质细胞生长因子和肝细胞生长因子， 体外肝细胞和体内正常肝脏， 阐明负责肝增殖的机制;和5）基因 转移可用于研究的生理和病理生理 肝脏的再生和葡萄糖代谢。 具体目标是 建议包括：1）分析载体效率、基因表达和基因表达的模式 表达和宿主免疫应答， 四氯化物处理的小鼠肝脏使用重组腺病毒， 标记基因; 2）研究核结合蛋白在 分离的小鼠肝细胞，正常肝，用重组转导的肝 腺病毒和再生肝; 3）研究腺病毒在肝再生中的作用。 抗凋亡因子Bcl-2和Bcl-XL，在延长的重组 腺病毒介导的基因转移在再生肝脏中观察到; 4）分析 用β-半乳糖苷酶转基因标记的肝细胞 在存在或不存在重组体的情况下植入小鼠肝脏 表达角质细胞生长因子和/或肝细胞生长的腺病毒 5）研究肝靶向重组腺病毒对肝癌细胞的作用 介导的葡萄糖激酶过表达对葡萄糖稳态和肝脏的影响 链脲佐菌素致糖尿病小鼠假手术或70% 肝切除

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.