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Quantitative Analysis of Cellular Composition in Advanced Atherosclerotic Lesions of Smooth Muscle Cell Lineage-Tracing Mice
Published on: February 20, 2019
Quantitative Analysis of Cellular Composition in Advanced Atherosclerotic Lesions of Smooth Muscle Cell
Sidney Mahan1, Mingjun Liu1, Richard A Baylis2
1Heart, Lung, Blood and Vascular Medicine Institute, University of Pittsburgh.
Abstract:
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.
Insights
Developing new treatments for atherosclerosis, the leading cause of death, requires better preclinical models. This study proposes advanced methods for analyzing cellular changes in established atherosclerotic lesions to improve therapeutic interventions.
Area of Science:
- Cardiovascular Biology
- Translational Medicine
- Atherosclerosis Research
Background:
- Atherosclerosis is a major global cause of mortality, yet effective therapeutic interventions remain limited.
- Current preclinical models often focus on disease prevention rather than established atherosclerosis, hindering translation to clinical practice.
- Superficial lesion analysis and lack of cellular characterization in preclinical studies confound results.
Purpose of the Study:
- To propose improved preclinical intervention models for atherosclerosis research.
- To advocate for single-cell level analysis of cellular composition in atherosclerotic lesions.
- To enhance the accuracy of therapeutic intervention modeling for cardiovascular disease.
Main Methods:
- Detailed protocol for testing therapeutic agents in a murine atherosclerosis intervention model.
- Systematic approach for animal dissection, tissue processing, and brachiocephalic artery lesion analysis.
- Utilizing immunofluorescent staining, confocal microscopy, and cell-specific inducible lineage tracing for unbiased cellular characterization.
Main Results:
- Established a comprehensive protocol for evaluating therapeutic interventions in established atherosclerotic lesions.
- Demonstrated the utility of single-cell analysis for characterizing diverse cell populations within lesions.
- Highlighted the importance of inducible lineage tracing for accurate assessment of cellular contributions.
Conclusions:
- Advanced preclinical models focusing on cellular composition at a single-cell level are crucial for overcoming translational hurdles in atherosclerosis research.
- Implementing these strategies can lead to more accurate modeling of therapeutic interventions.
- Improved preclinical modeling holds the potential to increase the success rate of clinical trials for atherosclerosis treatments.
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