Biliary oxysterols and cholangiocarcinoma

胆汁氧甾醇和胆管癌

• 批准号: 7574433
• 负责人: RAHUL P KUVER
• 金额: $ 21.24万
• 依托单位: SEATTLE INST FOR BIOMEDICAL/CLINICAL RES
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7574433
• 关键词: American Indians; Anchorage-Independent Growth; Apoptosis; Area; Asians; Autopsy; Bacteria; Bacterial Infections; Behavior; Bile fluid; Biliary; Biochemical; Biochemical Reaction; Biological; Biological Process; Calculi; Cancer cell line; Canis familiaris; Carcinogenesis Mechanism; Caucasians; Caucasoid Race; Cell Proliferation; Cell membrane; Cells; Cellular Morphology; Cellular biology; Characteristics; Chile; Chinese People; Cholangiocarcinoma; Cholangitis; Cholelithiasis; Cholesterol; Chronic; Clinical; Disease; Enzymes; Epidemiology; Epithelial; Epithelial Cells; Epithelium; Etiology; Event; Exhibits; Exposure to; Frequencies; Gallbladder; Growth; High Pressure Liquid Chromatography; Human; In Vitro; Incidence; Infection; Inflammatory; Intrahepatic Cholangiocarcinoma; Kinetics; Leukocytes; Lipids; Long-Term Effects; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of gallbladder; Maps; Mass Spectrum Analysis; Mediating; Membrane Microdomains; Metabolism; Methodology; Microbial Biofilms; Mitogen-Activated Protein Kinases; Modeling; Molecular; Molecular Bank; Molecular and Cellular Biology; Morphology; Mucins; Mummies; Native Americans; Nude Mice; Oryctolagus cuniculus; Pathologic Processes; Pathway interactions; Patients; Physical Chemistry; Physiology; Population; Prevalence; Prevention; Process; Production; Race; Recurrence; Sampling; Signal Transduction Pathway; Site; South American; South American Indians; Sterility; Sterols; Structural Biochemistry; Structure; Sum; Surface; Techniques; Time; United States National Institutes of Health; Work; biliary tract; carcinogenesis; cell growth; design; experience; in vivo; insight; interdisciplinary approach; intrahepatic; mortality; oxidation; primary sclerosing cholangitis; repository; research study; response; tumor

There is a worldwide and steady increase in the incidence of cholangiocarcinoma, a malignancy with very limited treatment options and high mortality. This expecially involves racial groups where the prevalence of biliary stones are high - such as Native Americans, South American Indians, andAsians. Of all the etiologic factors, cancers of the gallbladder and the biliary tree are most highly correlated with the presence of biliary stones, and chronic bacterial infection. We have recently discovered that in patients with gallbladder and intrahepatic stones, which are almost always covered by a bacterial biofilm, that there are oxidzed products of cholesterol. Two of these oxysterol species have been unequivocally identified. The presence and abundance of oxysterols in human bile and gallstones correlated with the presence of bacterial infection, and with the amount of bacterial DMA present in the stones. Oxysterols, on short term exposure to biliary epithelial cells, exhibit profound effects on apoptosis, cell proliferation, mucin synthesis and secretion. With long term exposure, dog gallbladder epithelial exhibited features characteristic of malignant transformation. We hypothesize that biliary oxysterols originate from endogenous biliary cholesterol as a result of oxidation from leukocytic enzymes in bacterial-leukocyte interactions. In addition, oxysterols mediate inflammatory and malignant transformation of biliary epithelial cells. We also hypothesizethat the molecular mechanism of oxysterol induced carcinogenesis to mediate through lipid microdomains (rafts). The Specific AimsTof this proposal are: 1) To determine the origin of biliary' bxysJerol.'We propose in vitro ' experiments using human leukocytes and model bile, as well,as in vivo experiments using a rabbit model with controlled induction of bacterial infection. The1 kinetics ofioxysterol production, the intermediates and the products wilhbe determined 2a) To construct a1 molecular library of;the;composition and structural identity of the oxidative products of cholesterol in hurpan bile'using high perforirfancdjiiiquidilchromatography (HPLC), interfacing,,with '''ij':^ j mass spectometry (MS) and, IMS/MS. 2$),To characterize'ithe'physicpchemical and biological function of'these ;|y', | oxysterols. 3) To use oxysterol in long-term co-cultur;e with normal human biliary (gallbladder) epithelial cells to'''/1 induce carciongenesis. Cell morphology/cancerjgrqwth pathways wiil.'be determined before,' du'ringiand after V cancer formation., _ __, 'jfv \t\ '¿, V'1' 'li'i !!'' JU'H . . '¿!;! :!''<'.! i"V ¿ ,'i1. ¿.! this proposal uses a multidisciplinary approach including structural biochemistry, cell and molecular biology, and physical chemistry methodologies. It promises to open up a new area of biliary physiology and metabolism; create a molecular library for cholesterol oxidation products in humans; add new and important understanding and insight into the etiology of cholangiocarcinoma; and guide strategic treatment and prevention of this important and vexinq disease.

胆管癌的发病率在全球范围内稳步上升，这是一种非常严重的恶性肿瘤 治疗选择有限，死亡率高。这尤其涉及到胆汁性疾病流行的种族群体 石头很高--比如美洲原住民、南美印第安人和亚洲人。在所有的病因中， 胆囊癌和胆管树癌与胆结石的存在高度相关，以及 慢性细菌感染。我们最近发现，在患有胆囊结石和肝内结石的患者中， 几乎总是被细菌生物膜覆盖，有胆固醇的氧化产物。其中两个 这些氧甾醇物种已经被明确地鉴定出来了。人体内氧化甾醇的存在和丰富 胆汁和胆结石与细菌感染的存在和细菌DNA的数量有关。 出现在石头里。氧化甾醇对胆管上皮细胞的短期暴露具有深远的影响 细胞凋亡、细胞增殖、粘蛋白合成和分泌。长期暴露，狗的胆囊腺上皮 表现出恶变的特征。 我们假设胆汁中的氧化甾醇来源于内源性胆汁胆固醇，其原因是胆汁中的 细菌-白细胞相互作用中的白细胞酶。此外，氧化甾醇还介导炎症和恶性反应。 胆管上皮细胞的转化。我们还假设，氧固醇诱导的分子机制 通过脂质微域(RAFT)介导的致癌作用。 这一建议的具体目的是：1)确定胆汁‘bxysJerol’的来源。我们建议在体外。 使用人类白细胞和模型胆汁的实验，以及使用兔模型的活体实验 控制细菌感染的诱导。氧化甾醇的生产、中间体和产物的动力学 将确定2a)以构建氧化剂的组成和结构同一性的A1分子文库 胆汁中胆固醇产物的高效液相色谱(HPLC法)，与 质谱仪(MS)和IMS/MS 2$)，以表征这些化合物的物理化学和生物功能；|y‘，| 氧类固醇。3)在长期共培养中使用氧化甾醇；与正常人胆管(胆)上皮细胞的比例为‘’/1 诱导癌变。细胞形态/癌变途径将在之前、之后和之后确定 癌症形成。，_，‘jfv\t\’，V‘1’‘li’i！‘’Ju‘h.。‘’！；！：！‘’&lt；‘.！I“V‘，’I1.‘！ 这项建议采用了包括结构生物化学、细胞和分子生物学在内的多学科方法， 物理化学方法论。它有望开辟胆道生理学和新陈代谢的新领域； 创建人类胆固醇氧化产物的分子库；增加新的和重要的理解和 深入了解胆管细胞癌的病因，并指导这一重要疾病的战略治疗和预防 维辛克病。