Novel heterotypic cell cultures for liver toxicology studies

用于肝毒理学研究的新型异型细胞培养物

• 批准号: 8323544
• 负责人: Alexander Revzin
• 金额: $ 18.29万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8323544
• 关键词: Adhesives; Adverse effects; Affect; Alcohol consumption; Alcohols; Attention; BMP7 gene; Caliber; Cell Culture System; Cell Culture Techniques; Cells; Coculture Techniques; Collagen; Complex; Development; Drug Metabolic Detoxication; Extracellular Matrix; Goals; Growth Factor; Hepatic; Hepatocyte; Hepatocyte Growth Factor; Human body; In Vitro; Injury; Lead; Liver; Liver Dysfunction; Liver Fibrosis; Location; Mediating; Metabolic; Mono-S; Phenotype; Play; Positioning Attribute; Principal Investigator; Printing; Process; Production; Rattus; Role; Signal Transduction; Signaling Molecule; Spottings; Technology; Testing; TimeLine; Toxicology; Transforming Growth Factors; Translating; alcohol effect; alcohol exposure; autocrine; cell type; high throughput screening; imprint; improved; in vivo Model; novel; paracrine; prevent; programs; receptor; research study; response; stellate cell; therapeutic target

DESCRIPTION (provided by applicant): The goal of our proposal is to investigate paracrine fibrogenic signaling between parenchymal and non parenchymal liver cells during alcohol injury. Whereas the nonparenchymalcompartment (stellate cells) remains the major player in the development of liver, the role of the parenchymal cells (hepatocytes) in this process is drawing increasing attention. Improved understanding of the roles played by parenchymal and nonparenchymalliver cells during alcohol insult will translate into better, more effective and targeted therapeutics. However, the complex signaling exchange between different liver cell types is difficult to delineate with in vivo models or standard in vitro cell culture approaches. In this proposal, we will utilize a novel liver cell culture approach to investigate !he hypothesis that hepatic production of transforming growth factor (TGF)-¿ plays an important role in initiating the fibrogneic program of stellate cells. This hypothesis will be explored using a novel cell culture system where cell adhesive and/or signaling molecules are imprinted into a culture substrate so as to position discrete groups of hepatocytes and stellate cells in defined locations and in close proximity to each other (see Figure 1 (A,B)). As shown in Figure 1 C, this novel cell culture dish will be used to selectively stimulate hepatocytes within the co-cultures with anti-fibrotic growth factors (GFs) (e.g. HGF and BMP7) during alcohol injury in order to investigate how GF signals delivered to hepatocytes impact activation of neighboring stellate cells. The novelty of the proposed platform lies in our ability to define interactions of two liver cell types cultured in the same dish so as to modulate the phenotype of one cell type and to study the response of the other cell type. This approach is particularly well-suited for improving our understanding of the complex heterotypic signaling underlying liver fibrosis. The novel cell culture system described here is envisioned as an enabling technology for liver toxicology studies and for high-throughput screening of liver-protective molecules.

描述(由申请人提供)：我们建议的目标是研究酒精损伤过程中实质和非实质肝细胞之间的旁分泌纤维形成信号。虽然非实质室(星状细胞)仍然是肝脏发育的主要参与者，但实质细胞(肝细胞)在这一过程中的作用正受到越来越多的关注。更好地理解实质细胞和非实质细胞在酒精侮辱中所起的作用将转化为更好、更有效和更有针对性的治疗方法。然而，不同类型的肝细胞之间复杂的信号交换很难用体内模型或标准的体外细胞培养方法来描述。在这项建议中，我们将利用一种新的肝细胞培养方法来研究这一假说，即肝脏产生转化生长因子(TGF)-β在启动星状细胞的纤维化程序中起着重要作用。这一假设将使用一种新型的细胞培养系统来探索，在该系统中，细胞黏附分子和/或信号分子被印记在培养底物中，以便将离散的肝细胞组和星状细胞组定位在指定的位置和彼此紧密接近的位置(见图1(A，B))。如图1C所示，这种新型细胞培养皿将用于在酒精损伤期间选择性地刺激与抗纤维化生长因子(GFS)(如HGF和BMP7)共培养的肝细胞，以研究传递给肝细胞的GF信号如何影响邻近星状细胞的激活。该平台的新奇之处在于，我们能够定义在同一培养皿中培养的两种肝细胞类型的相互作用，从而调节一种细胞类型的表型，并研究另一种细胞类型的反应。这种方法特别适合于提高我们对肝纤维化背后复杂的异型信号的理解。这里描述的新型细胞培养系统被设想为一种能够用于肝脏毒理学研究和高通量筛选肝脏保护分子的技术。