University Medical Center Groningen, Department of Pathology and Medical Biology, Medical Biology section, University of Groningen, Groningen, The Netherlands.
Cell Tissue Res. 2013 Dec;354(3):647-69. doi: 10.1007/s00441-013-1714-7. Epub 2013 Sep 27.
Many studies on the molecular control underlying normal cell behavior and cellular responses to disease stimuli and pharmacological intervention are conducted in single-cell culture systems, while the read-out of cellular engagement in disease and responsiveness to drugs in vivo is often based on overall tissue responses. As the majority of drugs under development aim to specifically interact with molecular targets in subsets of cells in complex tissues, this approach poses a major experimental discrepancy that prevents successful development of new therapeutics. In this review, we address the shortcomings of the use of artificial (single) cell systems and of whole tissue analyses in creating a better understanding of cell engagement in disease and of the true effects of drugs. We focus on microvascular endothelial cells that actively engage in a wide range of physiological and pathological processes. We propose a new strategy in which in vivo molecular control of cells is studied directly in the diseased endothelium instead of at a (far) distance from the site where drugs have to act, thereby accounting for tissue-controlled cell responses. The strategy uses laser microdissection-based enrichment of microvascular endothelium which, when combined with transcriptome and (phospho)proteome analyses, provides a factual view on their status in their complex microenvironment. Combining this with miniaturized sample handling using microfluidic devices enables handling the minute sample input that results from this strategy. The multidisciplinary approach proposed will enable compartmentalized analysis of cell behavior and drug effects in complex tissue to become widely implemented in daily biomedical research and drug development practice.
许多关于正常细胞行为和细胞对疾病刺激及药物干预反应的分子调控的研究都是在单细胞培养系统中进行的，而细胞在疾病中的参与和对药物的反应的读出通常基于整体组织反应。由于大多数正在开发的药物旨在与复杂组织中细胞亚群中的分子靶标特异性相互作用，这种方法存在一个主要的实验差异，从而阻止了新疗法的成功开发。在这篇综述中，我们讨论了在人工（单一）细胞系统和整个组织分析中使用的不足之处，以更好地了解细胞在疾病中的参与和药物的真实作用。我们专注于微血管内皮细胞，它们积极参与广泛的生理和病理过程。我们提出了一种新策略，即直接在病变的内皮细胞中研究体内细胞的分子调控，而不是在药物必须作用的部位之外，从而考虑到组织控制的细胞反应。该策略使用基于激光微切割的微血管内皮细胞富集，与转录组和（磷酸化）蛋白质组分析相结合，提供了它们在复杂微环境中的实际状态。将这与使用微流控装置进行微型化样本处理相结合，可处理该策略产生的微小样本输入。拟议的多学科方法将使复杂组织中细胞行为和药物作用的分区分析能够在日常的生物医学研究和药物开发实践中得到广泛实施。
