Detection and characterization of cell type-specific extracellular vesicles in obesity-driven hepatocellular carcinoma

肥胖导致的肝细胞癌中细胞类型特异性细胞外囊泡的检测和表征

• 批准号: 9806072
• 负责人: Takahisa Nakamura
• 金额: $ 18.5万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9806072
• 关键词: Affect; Biological Markers; Bone Marrow; Cell Culture Techniques; Cells; Characteristics; Complication; Detection; Development; Devices; Disease; Distant; Environment; Extracellular Space; Fatty acid glycerol esters; Genetic Transcription; Goals; Green Fluorescent Proteins; Hepatocyte; Immune; Immune Cell Activation; Inflammation; Inflammatory; Inflammatory Response; Interleukin-6; Label; Lead; Lipid Bilayers; Lipids; Liver; Liver Cirrhosis; Liver Fibrosis; Malignant Neoplasms; Mediating; Methods; Molecular; Monitor; Mus; Nucleic Acids; Obesity; Pathogenesis; Pathogenicity; Pathologic; Population; Prevalence; Primary carcinoma of the liver cells; Production; Proteins; Reproducibility; Risk Factors; Role; Serum; Sphingomyelinase; System; TNF gene; Technology; Time; Tissues; base; cancer cell; cancer risk; cell type; chronic liver inflammation; cytokine; dielectric property; exosome; extracellular vesicles; in vivo; in vivo monitoring; innovation; macrophage; microdevice; microvesicles; monocyte; mouse model; neutrophil; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; novel; pandemic disease; protein B; trait; tumor

PROJECT SUMMARY The worldwide prevalence of obesity has reached pandemic proportions. An association between fat accumulation or hepatosteatosis and hepatocellular carcinoma (HCC) development has been long known. Obesity-induced hepatosteatosis, together with its more severe complication nonalcoholic steatohepatitis (NASH), classified as the nonalcoholic fatty liver disease (NAFLD) affects up to 40% of the US population. Based on correlative and bench studies, several mechanisms have been proposed to explain how obesity increase cancer risk. An important finding that accounts for the tumor-promoting effect of obesity is the low- grade, aberrant inflammatory response, which results in the elevated production of cytokines, such as TNF and IL-6. Studies with mouse models demonstrated that obesity-promoted HCC development is dependent on the enhanced production of these inflammatory cytokines. However, critical questions concerning how the aberrant inflammation in obesity is initiated have been yet to be clearly defined. We hypothesize that in obesity, extracellular vesicles (EVs) derived from hepatocytes become pathogenic and drive recipient immune cells, such as neutrophils and monocytes/macrophages, towards abnormal inflammation associated with the development of HCC. To demonstrate this hypothesis, we will establish two novel systems; A) a new mouse line in which specific cell/tissue-derived EVs are selectively labeled with green fluorescent protein (GFP), and B) a simple, yet powerful electrokinetic-based micro-device that can rapidly extract EVs based on their dielectric properties from biofluids with high purity and yield. With these systems, we aim to 1) determine if H-EVs become pathogenic and induce aberrant inflammation leading to HCC, and 2) isolate H-EVs through the novel dielectrophoretic (DEP) technology and assess their pro- inflammatory trait. This study will lead us to demonstrate the pathogenicity of hepatocyte-derived EVs, which potentially provides a novel mechanism for the development of HCC and the EV biomarker for the disease.

项目摘要 肥胖症的全球流行率已经达到了大流行的比例。脂肪之间的关联 众所周知，积累或肝癌和肝细胞癌（HCC）发育已知。 肥胖引起的肝造成病，以及更严重的并发症非酒精性脂肪性肝炎 （NASH）被归类为非酒精性脂肪肝病（NAFLD）影响多达40％的美国人口。 基于纠正和台式研究，已经提出了几种机制来解释肥胖的方式 增加癌症风险。一个重要的发现，说明肥胖的肿瘤促进作用是低 - 等级，异常炎症反应，导致细胞因子的产生升高，例如TNF 和IL-6。使用小鼠模型的研究表明，肥胖的HCC发育取决于 这些炎症细胞因子的产生增强。但是，关于如何 启动肥胖的异常炎症尚未明确定义。 我们假设在肥胖症中，源自肝细胞的细胞外蔬菜（EV）成为致病性 并驱动接受者免疫球，例如中性粒细胞和单核细胞/巨噬细胞，趋向异常 与HCC的发展相关的炎症。为了证明这一假设，我们将建立两个 新型系统； a）特定细胞/组织衍生的EV选择性标记的新鼠标系 绿色荧光蛋白（GFP），b）一个简单但功能强大的基于电子的微型设备，可以 迅速提取电动汽车，其基于其纯净和产量的生物流体的顽固特性。与这些 系统，我们的目标是 1）确定H-EV是否成为致病性并诱导异常注射导致HCC，并且 2）通过新型的介电训练（DEP）技术分离H-EV，并评估其促进 炎症性状。 这项研究将使我们证明肝细胞衍生的电动汽车的致病性 HCC开发和EV生物标志物的新型机制。