Feenstra Lian, Zeper Lara W, van de Langenberg Brenda, Kahlman Eveline J E M, de La Roij Guido, Reijrink Melanie, Bernay Benoit, Chatre Laurent, Kuipers Jeroen, Giepmans Ben N G, Mastik Mirjam F, Kooistra Wierd, Lodewijk Monique E, Zuidscherwoude Malou, Pol Robert A, Smith Edward R, Krenning Guido, de Baaij Jeroen H F, Hillebrands Jan-Luuk, Hoenderop Joost G J
Department of Pathology and Medical Biology (HPC: EA10), University of Groningen, University Medical Center Groningen, Hanzeplein 1, 9713 GZ, Groningen, The Netherlands.
Department of Medical Biosciences, Radboud University Medical Center, P.O. Box 9101, 6500HB, Nijmegen, The Netherlands.
Cell Mol Life Sci. 2025 Apr 26;82(1):177. doi: 10.1007/s00018-025-05702-z.
Vascular calcification is highly prevalent in Chronic Kidney Disease (CKD) and is associated with markedly increased cardiovascular risk. High serum phosphate in CKD increases calcification propensity via generation of circulating calciprotein particles (CPP2), crystalline nanoaggregates composed of calcium, phosphate, and serum proteins. CPP2 induce vascular calcification in vascular smooth muscle cells (VSMCs) in vitro. In vivo, endothelial cells, rather than VSMCs are primarily exposed to CPP2, yet understanding the influence of endothelial cells on vascular calcification is limited.
We investigated calcification-promoting signalling by endothelial cells on VSMCs. Effects of CPP2 exposure to endothelial cells on CPP2 uptake, endothelial cell activation, and endothelial cell-derived secretome were studied. Effects of the secretome on VSMC calcification were investigated. Using NanoString nCounter analysis the effects of CPP2-activated endothelial cell-conditioned medium on VSMCs gene expression were mapped.
Endothelial cells internalise CPP2 and elevate ICAM-1, E-selectin, and VCAM-1-mRNA expression, indicating endothelial activation. VSMCs cultured in conditioned medium from CPP2-activated endothelial cells demonstrated enhanced calcification, suggesting that CPP2-activated endothelial cells release pro-calcifying soluble factors. Mass spectrometry was utilized to identify 1171 proteins in the CPP2-activated endothelial cells' secretome. Among these, 76 proteins were differentially expressed compared to control endothelial cells' secretome, including proteins related to blood vessel development, extracellular matrix remodelling, and oxidative stress-related processes. Finally, endothelial cell-derived paracrine factors present in conditioned medium enhanced mRNA-expression of calcification-related factors in VSMCs.
CPP2-activated endothelial cells promote VSMC calcification via paracrine signalling. In response to these paracrine factors, VSMCs increase the expression of pro-calcification genes.
血管钙化在慢性肾脏病（CKD）中非常普遍，并且与心血管风险显著增加相关。CKD患者的高血清磷通过生成循环钙蛋白颗粒（CPP2）增加钙化倾向，CPP2是由钙、磷和血清蛋白组成的结晶纳米聚集体。CPP2在体外可诱导血管平滑肌细胞（VSMC）发生血管钙化。在体内，主要是内皮细胞而非VSMC暴露于CPP2，但目前对内皮细胞在血管钙化中的作用了解有限。
我们研究了内皮细胞对VSMC的促钙化信号传导。研究了CPP2作用于内皮细胞对CPP2摄取、内皮细胞活化及内皮细胞分泌组的影响。研究了分泌组对VSMC钙化的影响。使用NanoString nCounter分析绘制了CPP2激活的内皮细胞条件培养基对VSMC基因表达的影响。
内皮细胞摄取CPP2并提高细胞间黏附分子-1（ICAM-1）、E-选择素和血管细胞黏附分子-1（VCAM-1）的mRNA表达，表明内皮细胞被激活。在CPP2激活的内皮细胞条件培养基中培养的VSMC显示钙化增强，提示CPP2激活的内皮细胞释放促钙化可溶性因子。利用质谱鉴定了CPP2激活的内皮细胞分泌组中的1171种蛋白质。其中，与对照内皮细胞分泌组相比，有76种蛋白质差异表达，包括与血管发育、细胞外基质重塑和氧化应激相关过程的蛋白质。最后，条件培养基中存在的内皮细胞衍生旁分泌因子增强了VSMC中钙化相关因子的mRNA表达。
CPP2激活的内皮细胞通过旁分泌信号传导促进VSMC钙化。VSMC对这些旁分泌因子作出反应后，会增加促钙化基因的表达。
