Department of Bioengineering, University of California, San Diego, USA.
Integr Biol (Camb). 2009 Sep;1(8-9):513-24. doi: 10.1039/b912926j. Epub 2009 Aug 10.
Hepatic stellate cells (HSCs) are a major cell type of the liver that are involved in liver homeostasis. Upon liver damage, HSCs exit their normally quiescent state and become activated, leading to an increase of their proliferation, production of abnormal extracellular matrix proteins (ECMPs) and inflammatory mediators, and eventually liver fibrosis and cirrhosis. Current in vitro approaches to identify components that influence HSC biology typically investigate one factor at a time and generally ignore the complex crosstalk among the myriad of components that comprise the microenvironments of quiescent or activated HSCs. Here we describe a high throughput screening (HTS) approach to identify factors that affect HSC biology. Specifically, we integrated the use of ECMPs and signaling molecules into a combinatorial cellular microarray technology platform, thereby creating comprehensive "microenvironments". Using this technology, we performed real-time simultaneous screening of the effects of hundreds of unique microenvironments composed of ECMPs and signaling molecules on HSC proliferation and activation. From these screens, we identified combinations of microenvironment components that differentially modulate the HSC phenotype. Furthermore, analysis of HSC responses revealed that the influences of Wnt signaling molecules on HSC fate are dependent on the ECMP composition in which they are presented. Collectively, our results demonstrate the utility of high-content, array-based screens to provide a better understanding of HSC biology. Our results indicate that array-based screens may provide an efficient means for identifying candidate signaling pathways to be targeted for anti-fibrotic therapies.
肝星状细胞（HSCs）是肝脏的主要细胞类型之一，参与肝脏稳态。在肝损伤时，HSCs 从正常静止状态中退出并被激活，导致其增殖增加、产生异常细胞外基质蛋白（ECMPs）和炎症介质，最终导致肝纤维化和肝硬化。目前用于鉴定影响 HSC 生物学的成分的体外方法通常一次研究一个因素，并且通常忽略了构成静止或激活 HSCs 微环境的无数成分之间的复杂串扰。在这里，我们描述了一种用于鉴定影响 HSC 生物学的因素的高通量筛选（HTS）方法。具体来说，我们将 ECMPs 和信号分子的使用整合到组合细胞微阵列技术平台中，从而创建了全面的“微环境”。使用这项技术，我们实时同时筛选了由 ECMPs 和信号分子组成的数百种独特微环境对 HSC 增殖和激活的影响。通过这些筛选，我们鉴定了可以差异调节 HSC 表型的微环境成分组合。此外，对 HSC 反应的分析表明，Wnt 信号分子对 HSC 命运的影响取决于它们呈现的 ECMP 组成。总的来说，我们的结果证明了基于高内涵、基于阵列的筛选在更好地理解 HSC 生物学方面的实用性。我们的结果表明，基于阵列的筛选可能为鉴定抗纤维化治疗的候选信号通路提供一种有效的手段。
