Department of Pathology and Laboratory Medicine University of British Columbia Vancouver BC Canada.
Centre for Heart Lung Innovation University of British Columbia Vancouver BC Canada.
J Am Heart Assoc. 2024 Jun 18;13(12):e034990. doi: 10.1161/JAHA.123.034990. Epub 2024 Jun 6.
Previous studies using animal models and cultured cells suggest that vascular smooth muscle cells (SMCs) and inflammatory cytokines are important players in atherogenesis. Validating these findings in human disease is critical to designing therapeutics that target these components. Multiplex imaging is a powerful tool for characterizing cell phenotypes and microenvironments using biobanked human tissue sections. However, this technology has not been applied to human atherosclerotic lesions and needs to first be customized and validated.
For validation, we created an 8-plex imaging panel to distinguish foam cells from SMC and leukocyte origins on tissue sections of early human atherosclerotic lesions (n=9). The spatial distribution and characteristics of these foam cells were further analyzed to test the association between SMC phenotypes and inflammation. Consistent with previous reports using human lesions, multiplex imaging showed that foam cells of SMC origin outnumbered those of leukocyte origin and were enriched in the deep intima, where the lipids accumulate in early atherogenesis. This new technology also found that apoptosis or the expression of pro-inflammatory cytokines were not more associated with foam cells than with nonfoam cells in early human lesions. More CD68 SMCs were present among SMCs that highly expressed interleukin-1β. Highly inflamed SMCs showed a trend of increased apoptosis, whereas leukocytes expressing similar levels of cytokines were enriched in regions of extracellular matrix remodeling.
The multiplex imaging method can be applied to biobanked human tissue sections to enable proof-of-concept studies and validate theories based on animal models and cultured cells.
先前使用动物模型和培养细胞的研究表明，血管平滑肌细胞（SMC）和炎症细胞因子是动脉粥样硬化形成过程中的重要参与者。在人类疾病中验证这些发现对于设计针对这些成分的治疗方法至关重要。多重免疫荧光成像技术是一种使用生物样本库的人类组织切片来描述细胞表型和微环境的强大工具。然而，该技术尚未应用于人类动脉粥样硬化病变，需要首先进行定制和验证。
为了进行验证，我们创建了一个 8 重免疫荧光成像面板，以区分组织切片中早期人类动脉粥样硬化病变中的泡沫细胞来源于 SMC 和白细胞（n=9）。进一步分析这些泡沫细胞的空间分布和特征，以测试 SMC 表型和炎症之间的关联。与先前使用人类病变的报告一致，多重免疫荧光成像显示，SMC 来源的泡沫细胞数量多于白细胞来源的泡沫细胞，并且在深层内膜中富集，脂质在早期动脉粥样硬化形成过程中积累。这项新技术还发现，在早期人类病变中，与非泡沫细胞相比，细胞凋亡或促炎细胞因子的表达与泡沫细胞的相关性并不更强。在高表达白细胞介素 1β的 SMC 中，存在更多的 CD68 SMC。高度炎症的 SMC 表现出凋亡增加的趋势，而表达类似细胞因子水平的白细胞则富集在细胞外基质重塑区域。
多重免疫荧光成像方法可以应用于生物样本库的人类组织切片，以进行概念验证研究并验证基于动物模型和培养细胞的理论。
