Quantitative Analysis of Cellular Composition in Advanced Atherosclerotic Lesions of Smooth Muscle Cell Lineage-Tracing Mice.
Quantitative Analysis of Cellular Composition in Advanced Atherosclerotic Lesions of Smooth Muscle Cell Lineage-Tracing Mice.
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DOI：
10.3791/59139
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发表时间：
2019-02
期刊：
影响因子：
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通讯作者：
Sidney Mahan;Mingjun Liu;Richard A. Baylis;Delphine Gomez
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文献类型：
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作者：
Sidney Mahan;Mingjun Liu;Richard A. Baylis;Delphine Gomez
动脉粥样硬化仍然是全球死亡的主要原因，尽管有无数的临床前研究描述了有希望的治疗靶点，但新的干预措施仍然难以捉摸。这可能部分是由于依赖于临床前预防模型，研究基因操作或药物治疗对动脉粥样硬化发展的影响，而不是确定的疾病。此外，由于使用浅表病变分析和缺乏病变细胞群的表征，这些研究的结果通常是混淆的。为了帮助克服这些翻译障碍，我们建议增加对干预模型的依赖，这些模型通过免疫荧光染色和共聚焦显微镜在单细胞水平上研究细胞组成的变化。为此，我们描述了一个协议，用于测试一个假定的治疗剂在小鼠干预模型，包括动物解剖，包埋，切片，染色和定量头臂动脉病变的系统方法。此外，由于晚期动脉粥样硬化病变内细胞的表型多样性，我们描述了使用细胞特异性，诱导谱系示踪小鼠系统的重要性，以及如何利用这一点对动脉粥样硬化病变细胞群进行无偏表征。总之，这些策略可以帮助血管生物学家更准确地模拟治疗干预和分析动脉粥样硬化疾病，并有望转化为更高的临床试验成功率。
Atherosclerosis remains the leading cause of death worldwide and, despite countless preclinical studies describing promising therapeutic targets, novel interventions have remained elusive. This is likely due, in part, to a reliance on preclinical prevention models investigating the effects of genetic manipulations or pharmacological treatments on atherosclerosis development rather than the established disease. Also, results of these studies are often confounding because of the use of superficial lesion analyses and a lack of characterization of lesion cell populations. To help overcome these translational hurdles, we propose an increased reliance on intervention models that employ investigation of changes in cellular composition at a single cell level by immunofluorescent staining and confocal microscopy. To this end, we describe a protocol for testing a putative therapeutic agent in a murine intervention model including a systematic approach for animal dissection, embedding, sectioning, staining, and quantification of brachiocephalic artery lesions. In addition, due to the phenotypic diversity of cells within late-stage atherosclerotic lesions, we describe the importance of using cell-specific, inducible lineage tracing mouse systems and how this can be leveraged for unbiased characterization of atherosclerotic lesion cell populations. Together, these strategies may assist vascular biologists to more accurately model therapeutic interventions and analyze atherosclerotic disease and will hopefully translate into a higher rate of success in clinical trials.