Biliary oxysterols and cholangiocarcinoma

胆汁氧甾醇和胆管癌

• 批准号: 7343156
• 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/7343156
• 关键词: American; American Indians; Anchorage-Independent Growth; Apoptosis; Area; Asians; Autopsy; Bacteria; Bacterial Infections; Behavior; Bile fluid; Biliary; Biliary calculi; 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; Condition; Disease; Enzymes; Epidemiology; Epithelial; Epithelial Cells; Epithelium; Etiology; Event; Exhibits; Exposure to; Frequencies; Gallbladder; Gallbladder Carcinoma; 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; Prostaglandin-Endoperoxide Synthase; Race; Recurrence; Sampling; Signal Transduction Pathway; Site; South American Indians; Sterility; Sterols; Structural Biochemistry; Structure; Sum; Surface; Techniques; Time; United States National Institutes of Health; Work; biliary tract; carcinogenesis; cell growth; cyclooxygenase 1; 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.

胆管癌的发病率在世界范围内稳步增加，胆管癌是一种恶性肿瘤， 有限的治疗选择和高死亡率。这尤其涉及到胆道疾病患病率高的种族群体， 石头是高的-如美洲原住民，南美印第安人，和亚洲人。在所有的病因中， 胆囊癌和胆道系统癌与胆结石的存在最密切相关， 慢性细菌感染我们最近发现，在胆囊和肝内结石患者中， 它们几乎总是被细菌的生物膜覆盖，其中有胆固醇的氧化产物。两 这些氧化甾醇种类已被明确鉴定。人体内氧固醇的存在和丰度 胆汁和胆结石与细菌感染的存在和细菌DMA的量相关 存在于石头中。氧化固醇，在短期暴露于胆管上皮细胞，表现出深刻的影响， 细胞凋亡、细胞增殖、粘蛋白合成和分泌。长期暴露，狗胆囊上皮细胞 表现出恶性转化的特征。 我们假设胆汁氧化固醇来源于内源性胆汁胆固醇， 细菌-白细胞相互作用中的白细胞酶。此外，氧固醇介导炎症和恶性 胆管上皮细胞的转化。我们还推测氧化固醇诱导的分子机制 通过脂质微区（筏）介导致癌。 本研究的具体目的是：1）确定胆固醇的来源。'我们建议在体外' 使用人白细胞和模型胆汁的实验，以及使用兔模型的体内实验， 控制细菌感染的诱导。氧化甾醇的生产动力学、中间体和产物 2a）构建了一个含有氧化还原酶的组成和结构特性的分子文库， 使用高效液相色谱法（HPLC）测定肝胆汁中的胆固醇产物，与“ij”接口： 质谱（MS）和IMS/MS 2$），以表征这些物质的物理化学和生物学功能;| y '，| 氧化固醇3)使用氧化固醇与正常人胆管（胆囊）上皮细胞长期共培养，以“"/1 诱发癌变。细胞形态学/肿瘤生长途径。在V之前，在V之后， 癌症形成，_，'jfv \t\ '<$，V'1' 'li'i！！Ju 'H。. '¿！;！:! ''<'.! i“V "i 1.¿.！ 该建议使用多学科方法，包括结构生物化学，细胞和分子生物学， 和物理化学方法。它有望开辟胆道生理学和代谢学的新领域; 创建人体胆固醇氧化产物的分子库;增加新的和重要的理解， 深入了解胆管癌的病因;并指导战略治疗和预防这一重要和 vexinq病。