Genesis of Liver Carcinomas with Oval Cell Traits

具有卵圆细胞特征的肝癌的起源

• 批准号: 7483693
• 负责人: Douglas Carter Hixson
• 金额: $ 33.86万
• 依托单位: RHODE ISLAND HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7483693
• 关键词: Address; Adult; Adverse effects; Agar; Aneuploidy; Basal Cell; Bile Duct Epithelium; Biliary; Biological Assay; Biological Models; Birth; Blast Cell; Carcinogens; Carcinoma; Cell Death; Cell Proliferation; Cell division; Cells; Characteristics; Chemical Injury; Cholangiocarcinoma; Choline; Chromosomes; Chronic; Complex; Development; Diet; Diethylnitrosamine; Dipeptidyl-Peptidase IV; Duct (organ) structure; Ductal; Ductal Epithelial Cell; Epithelial Cells; Exposure to; Fetal Liver; Furans; Genomics; Growth; Hepatic; Hepatitis B Virus; Hepatitis C; Hepatocarcinogenesis; Hepatocyte; Human; In Situ; In Vitro; Invasive; Kinetochores; Lead; Learning; Liver; Liver Regeneration; Malignant neoplasm of liver; Maps; Membrane; Mesenchyme; Methodology; Microtubule-Associated Proteins; Microtubules; Mitomycin; Modeling; Molecular; Monoclonal Antibodies; Newborn Infant; Pancreas; Partial Hepatectomy; Patients; Plastics; Pregnancy; Primary carcinoma of the liver cells; Probability; Proteins; Proteomics; Rattus; Reporting; Retroviridae; Role; Scheme; Signal Pathway; Staging; Stem cells; Structure of intralobular bile duct; Testing; Transfection; Transplantation; Work; base; biliary tract; carcinogenesis; cell type; cholangiocyte; experience; fetal; furan; hydroxy-aluminum polymer; in vivo; insight; interest; intrahepatic; novel; oval cell; progenitor; repaired; research study; retrorsine; trait

DESCRIPTION (provided by applicant): Although there is compelling evidence for the presence of bipotent progenitor cells in the adult liver, the role of these progenitors s in hepatocarcinogenesis is still a subject of debate. Building on our past experience with cholangiocytes and oval cells, bipotent biliary progenitors activated by most hepatocarcinogens, we have continued with our efforts to identify and characterize bipotent progenitors present in the biliary tree and to ascertain their role in hepato-and cholangio-carcinogenesis. Over the past 4 years, our studies of cholangiocyte marker positive (CMP), bipotent, fetal liver epithelial cells (FLEC) have yielded novel monoclonal antibody based schemes for isolating bipotent CMP-FLEC. Results from transplantation of CMP-FLEC isolates from dipeptidyl peptidase IV (DPPIV) positive rats into DPPIV deficient host rats treated with retrorsine/partial hepatectomy (R/PH) led to the unexpected finding that CMP-FLEC possesses a much higher capacity for growth in the R/PH treated adult liver than fetal hepatoblasts and other nonparenchymal cell types. In the current proposal, isolation schemes developed for bipotent CMP-FLEC will be applied to the isolation of phenotypically equivalent CMP-liver epithelial cells (CMP-LEC) from newborn and adult rat liver. We hypothesize that these fetal-like CMP-FLEC will possess a capacity for hepatocytic differentiation similar to oval cells and will retain this capacity following spontaneous transformation in vitro and progression to HCC in vivo. In Specific Aim 1, we will employ a rapid transplantation model that replaces retrorsine with mitomycin C (mitoC/PH) to test the hypothesis that the expression of the cholangiocyte marker OC.4, a marker first seen at 2 after birth, identifies mature CMP-LEC that have a greatly diminished capacity for hepatocytic differentiation. In Specific Aim 2, we will test the hypothesis that spontaneously transformed CMP-LEC and oval cells but not BDEC will undergo incomplete hepatocytic differentiation in mitoC/PH treated rats and progress to CMP-HCC. Spontaneous transformation of CMP-LEC will be accelerated by selection on plastic and/or soft agar, transfection with ErbB2 or exposure to carcinogen in situ. In Specific Aim 3, genomic and proteomic methodologies will be used to map the signaling pathways operative during the spontaneous transformation of BDEC, CMP-LEC and oval cells that promote survival and cooperate with ErbB2 to confer anchorage independent/invasive growth. Specific Aim 4 will continue with the characterization of BD.1, a 170 kDa protein expressed by cholangiocytes but not oval cells that forms stable complexes with CLIP170, a microtubule associated protein. We will test the hypothesis that loss of BD.1 alters microtubule/kinetochore dynamics in a manner that promotes aneuploidy. The proposed studies will provide new insights into the role of bipotent progenitors in liver cancer.

描述（由申请人提供）：尽管有令人信服的证据表明成人肝脏中存在双能祖细胞，但这些祖细胞在肝癌发生中的作用仍然是一个争论的话题。基于我们过去对胆管细胞和卵圆细胞（大多数肝癌物质激活的双能胆管祖细胞）的经验，我们继续努力识别和表征胆管树中存在的双能祖细胞，并确定它们在肝癌和胆管癌发生中的作用。在过去的 4 年里，我们对胆管细胞标记物阳性 (CMP)、双能胎儿肝上皮细胞 (FLEC) 的研究产生了基于单克隆抗体的新型方案，用于分离双能 CMP-FLEC。将来自二肽基肽酶 IV (DPPIV) 阳性大鼠的 CMP-FLEC 分离物移植到经逆转录酶/部分肝切除术 (R/PH) 处理的 DPPIV 缺陷宿主大鼠中，结果意外地发现 CMP-FLEC 在 R/PH 处理的成年肝脏中比胎儿成肝细胞和其他非实质细胞类型具有更高的生长能力。在当前的提案中，为双能 CMP-FLEC 开发的分离方案将应用于从新生和成年大鼠肝脏中分离表型等效的 CMP 肝上皮细胞 (CMP-LEC)。我们假设这些胎儿样 CMP-FLEC 将具有类似于卵圆细胞的肝细胞分化能力，并且在体外自发转化和体内进展为 HCC 后将保留这种能力。在具体目标 1 中，我们将采用一种用丝裂霉素 C (mitoC/PH) 替代逆转录碱的快速移植模型来检验以下假设：胆管细胞标记物 OC.4（出生后 2 岁时首次出现的标记物）的表达可识别肝细胞分化能力大大降低的成熟 CMP-LEC。在具体目标 2 中，我们将测试以下假设：自发转化的 CMP-LEC 和卵圆细胞（而非 BDEC）将在 mitoC/PH 治疗的大鼠中经历不完全肝细胞分化并进展为 CMP-HCC。通过塑料和/或软琼脂上的选择、ErbB2 转染或原位暴露于致癌物质，将加速 CMP-LEC 的自发转化。在具体目标 3 中，基因组和蛋白质组学方法将用于绘制 BDEC、CMP-LEC 和卵圆细胞自发转化过程中起作用的信号通路，这些细胞促进存活并与 ErbB2 配合以赋予贴壁独立/侵入性生长。具体目标 4 将继续表征 BD.1，BD.1 是一种由胆管细胞而非卵圆细胞表达的 170 kDa 蛋白，与微管相关蛋白 CLIP170 形成稳定的复合物。我们将检验这样的假设：BD.1 的缺失会以促进非整倍性的方式改变微管/动粒动力学。拟议的研究将为双能祖细胞在肝癌中的作用提供新的见解。