Severe aortic stenosis (AS) is prevalent in adults ≥ 65 years, a significant cause of morbidity and mortality, with no medical therapy. Lipid and proteomic alterations of human AS tissue were determined using mass spectrometry imaging (MSI) and liquid chromatography electrospray ionization tandem mass spectrometry (LC–ESI–MS/MS) to understand histopathology, potential biomarkers of disease, and progression from non-calcified to calcified phenotype. A reproducible MSI method was developed using healthy murine aortic valves (n = 3) and subsequently applied to human AS (n = 2). Relative lipid levels were spatially mapped and associated with different microdomains. Proteomics for non-calcified and calcified microdomains were performed to ascertain differences in expression. Increased pro-osteogenic and inflammatory lysophosphatidylcholine (LPC) 16:0 and 18:0 were co-localized with calcified microdomains. Proteomics analysis identified differential patterns in calcified microdomains with high LPC and low cholesterol as compared to non-calcified microdomains with low LPC and high cholesterol. Calcified microdomains had higher levels of: apolipoproteins (Apo) B-100 (p < 0.001) and Apo A-IV (p < 0.001), complement C3 and C4-B (p < 0.001), C5 (p = 0.007), C8 beta chain (p = 0.013) and C9 (p = 0.010), antithrombotic proteins alpha-2-macroglobulin (p < 0.0001) and antithrombin III (p = 0.002), and higher anti-calcific fetuin-A (p = 0.02), while the osteoblast differentiating factor transgelin (p < 0.0001), extracellular matrix proteins versican, prolargin, and lumican ( p < 0.001) and regulator protein complement factor H (p < 0.001) were higher in non-calcified microdomains. A combined lipidomic and proteomic approach provided insight into factors potentially contributing to progression from non-calcified to calcific disease in severe AS. Additional studies of these candidates and protein networks could yield new targets for slowing progression of AS.

严重主动脉瓣狭窄 (AS) 在 65 岁以上的成年人中普遍存在，这是发病率和死亡率的重要原因，没有药物治疗。使用质谱成像 (MSI) 和液相色谱电喷雾电离串联质谱 (LC-ESI-MS/MS) 确定人 AS 组织的脂质和蛋白质组学改变，以了解组织病理学、疾病的潜在生物标志物以及从非钙化到钙化的进展。钙化表型。使用健康的小鼠主动脉瓣 (n = 3) 开发了可重复的 MSI 方法，随后应用于人类 AS (n = 2)。相对脂质水平在空间上映射并与不同的微区相关联。进行非钙化和钙化微区的蛋白质组学以确定表达的差异。增加促骨和炎性溶血磷脂酰胆碱 (LPC) 16：0 和 18:0 与钙化微区共定位。蛋白质组学分析确定了具有高 LPC 和低胆固醇的钙化微区与具有低 LPC 和高胆固醇的非钙化微区的差异模式。钙化微区具有更高水平：载脂蛋白 (Apo) B-100 (p < 0.001) 和 Apo A-IV (p < 0.001)、补体 C3 和 C4-B (p < 0.001)、C5 (p = 0.007)、C8 β 链 (p = 0.013) 和 C9 (p = 0.010)，抗血栓蛋白 α-2-巨球蛋白 (p < 0.0001) 和抗凝血酶 III (p = 0.002)，以及更高的抗钙化胎球蛋白-A (p = 0.02)，而成骨细胞分化因子转凝胶蛋白 (p < 0.0001)、细胞外基质蛋白 versican、prolargin 和 lumican (p < 0.001) 和调节蛋白补体因子 H (p < 0.001) 在非钙化微区中较高。脂质组学和蛋白质组学相结合的方法提供了对可能导致严重 AS 从非钙化疾病进展为钙化疾病的因素的深入了解。对这些候选物和蛋白质网络的进一步研究可以产生减缓 AS 进展的新目标。