Human Breast Tumor-associated Endothelial Cells Favor Interactions with Nanomedicines

Nanomedicines, designed to improve the effectiveness of cancer treatment, have not met anticipated outcomes owing to suboptimal delivery efficiency to tumors. While the enhanced permeability and retention (EPR) effect has been explored, recent research challenges the predominant passive accumulation theory. This study explores the mechanisms governing nanoparticle entry into tumors by comparing human breast tumor-associated endothelial cells (HBTECs) and healthy human mammary macrovascular endothelial cells (HMMECs). Our results demonstrate that HBTECs exhibit higher nanoparticle uptake, involving three major endocytic pathways. Gene expression analysis reveals the upregulation of transport-related genes, especially motor protein genes, in tumor-associated endothelial cells. These insights highlight the need for comprehensive investigations into nanoparticle transport mechanisms and associated factors within the tumor microenvironment, essential for advancing drug delivery and therapeutic outcomes. Overall design: To investigate the difference in nanoparticle interactions with endothelial cells from tumor and healthy tissues, we employed two primary human cell types: human breast tumor-associated endothelial cells (HBTECs) and human mammary microvascular endothelial cells (HMMECs). Two experimental conditions, one without and one with gold nanoparticle incubation, were applied. Consequently, we established four distinct groups: HBTEC, HMMEC, HBTEC_Au (HBTECs with gold nanoparticle incubation), and HMMEC_Au (HMMECs with gold nanoparticle incubation). RNA-seq was conducted on these groups. For the RNA-seq result analysis, the four groups were compared in a pairwise manner, which are HBTEC vs HMMEC, HBTEC_Au vs HMMEC_Au, HBTEC_Au vs HBTEC, and HMMEC_Au vs HMMEC, to explore the impact of both cell origin differences and nanoparticle incubation.

旨在提升癌症治疗效果的纳米药物，因向肿瘤递送的效率欠佳，尚未达到预期疗效。尽管人们已对增强渗透滞留效应（enhanced permeability and retention, EPR）展开研究，但近期研究对主流的被动积累理论提出了质疑。本研究通过对比人乳腺肿瘤相关内皮细胞（human breast tumor-associated endothelial cells, HBTECs）与健康人乳腺大血管内皮细胞（human mammary macrovascular endothelial cells, HMMECs），探究调控纳米颗粒进入肿瘤的机制。研究结果显示，HBTECs的纳米颗粒摄取能力更强，且涉及三大主要内吞通路。基因表达分析表明，肿瘤相关内皮细胞中与转运相关的基因（尤其是动力蛋白基因）表达上调。这些研究结果凸显了全面探究肿瘤微环境内纳米颗粒转运机制及其相关影响因素的必要性，而这对于优化药物递送与治疗效果至关重要。
实验整体设计：为探究纳米颗粒与肿瘤来源及健康组织来源内皮细胞的相互作用差异，本研究采用了两种原代人源细胞系：人乳腺肿瘤相关内皮细胞（HBTECs）与人乳腺微血管内皮细胞（human mammary microvascular endothelial cells, HMMECs）。设置两种实验条件：分别为未添加金纳米颗粒孵育与添加金纳米颗粒孵育。由此构建了四组独立样本：HBTEC组、HMMEC组、HBTEC_Au组（经金纳米颗粒孵育的HBTECs）以及HMMEC_Au组（经金纳米颗粒孵育的HMMECs）。对上述四组样本开展RNA测序（RNA-seq）实验。在RNA-seq结果分析阶段，以两两配对的方式对四组样本进行比较，即HBTEC组 vs HMMEC组、HBTEC_Au组 vs HMMEC_Au组、HBTEC_Au组 vs HBTEC组以及HMMEC_Au组 vs HMMEC组，以此探究细胞来源差异与金纳米颗粒孵育分别产生的影响。