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: 8073450
• 负责人: DAVID MILLS
• 金额: $ 35.31万
• 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 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8073450
• 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; Kinetochores; Lead; Learning; Liver; Liver Regeneration; Malignant neoplasm of liver; Maps; Membrane; Mesenchyme; Methodology; Microtubule-Associated Proteins; Microtubules; Mitomycins; 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 transformation; cell type; cholangiocyte; experience; fetal; in vivo; insight; interest; intrahepatic; novel; oval cell; progenitor; regenerative; 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二生性祖细胞，但这些祖细胞在肝癌发生中的作用仍然是争论的主题。在我们过去在胆管细胞和椭圆形细胞上的经验的基础上，大多数肝癌激活的双能胆道祖细胞激活，我们一直在努力识别和表征胆道树中存在的二聚体祖细胞并确定它们在肝癌和胆管中的作用。在过去的四年中，我们对胆管细胞标记阳性（CMP），二足，胎儿肝上皮细胞（FLEC）的研究产生了新型的基于单克隆抗体的方案，用于分离双能CMP-FLEC。从二肽基肽酶IV（DPPIV）阳性大鼠中CMP-FLEC分离株移植到DPPIV缺陷的宿主大鼠中，该大鼠接受了反性/部分肝切除术（R/pH），这导致了意外的发现，CMP-FLEC具有在CMP-FLEC中具有更高的增长能力。 R/pH治疗的成人肝脏比胎儿肝细胞和其他非核细胞类型。在当前的提案中，为二型CMP-FLEC开发的分离方案将应用于从新生儿和成年大鼠肝脏中分离出表型等效的CMP肝上皮细胞（CMP-LEC）。我们假设这些类似胎儿的CMP-FLEC将具有与椭圆形细胞相似的肝细胞分化能力，并且在体外自发转化和体内自发转化后将保持这种能力。在特定目标1中，我们将采用快速移植模型，该模型用丝霉素C（mitoc/pH）取代重试，以测试胆管细胞标记OC.4的表达的假设，即出生后首次看到的标记物（第2个标记），确定成熟的CMP CMP -LEC的肝细胞分化能力大大降低。在特定的目标2中，我们将测试以下假设：自发转化CMP-LEC和椭圆形细胞，而BDEC不会在MITOC/pH处理的大鼠中经历不完全的肝细胞分化并发展为CMP-HCC。 CMP-LEC的自发转化将通过对塑料和/或软琼脂的选择加速，用ERBB2转染或原位接触致癌物。在特定的目标3中，将使用基因组和蛋白质组学方法来绘制在BDEC，CMP-LEC和椭圆形细胞自发转化期间的信号传导途径，以促进生存并与ERBB2合作以赋予锚定独立/侵入性生长。特定的目标4将继续进行BD.1的表征，BD.1是一种由胆管细胞表达的170 kDa蛋白，而不是椭圆形细胞，该蛋白与Clip170（一种微管相关蛋白）形成稳定的复合物。我们将检验以下假设：BD的损失以促进非整倍性的方式改变微管/动力学动力学。拟议的研究将为二元祖细胞在肝癌中的作用提供新的见解。

项目成果

Cholangiocyte marker-positive and -negative fetal liver cells differ significantly in their ability to regenerate the livers of adult rats exposed to retrorsine.

胆管细胞标记物阳性和阴性的胎儿肝细胞在暴露于逆转录碱的成年大鼠的肝脏再生能力方面存在显着差异。

• DOI: 10.1242/dev.02589
• 发表时间: 2006
• 期刊: Development (Cambridge, England)
• 作者: Simper-Ronan,Rhonda;Brilliant,Kate;Flanagan,Donna;Carreiro,Marie;Callanan,Helen;Sabo,Edmond;Hixson,DouglasC
• 通讯作者: Hixson,DouglasC