Mueller Mikala C, Blomberg Rachel, Tanneberger Alicia E, Davis-Hall Duncan, Neeves Keith B, Magin Chelsea M
bioRxiv. 2025 Jan 21:2025.01.17.633670. doi: 10.1101/2025.01.17.633670.
Pulmonary arterial hypertension (PAH) impacts male and female patients in different ways. Female patients exhibit a greater susceptibility to disease (4:1 female-to-male ratio) but live longer after diagnosis than male patients. This complex sexual dimorphism is known as the estrogen paradox. Prior studies suggest that estrogen signaling may be pathologic in the pulmonary vasculature and protective in the heart, yet the mechanisms underlying these sex-differences in PAH remain unclear. PAH is a form of a pulmonary vascular disease that results in scarring of the small blood vessels, leading to impaired blood flow and increased blood pressure. Over time, this increase in blood pressure causes damage to the heart. Many previous studies of PAH relied on male cells or cells of undisclosed origin for modeling. Here we present a dynamic, 3D-bioprinted model that incorporates cells and circulating sex hormones from female patients to specifically study how female patients respond to changes in microenvironmental stiffness and sex hormone signaling. Poly(ethylene glycol)-alpha methacrylate (PEGαMA)-based hydrogels containing female human pulmonary artery adventitia fibroblasts (hPAAFs) from idiopathic PAH (IPAH) or control donors were 3D bioprinted to mimic pulmonary artery adventitia. These biomaterials were initially soft, like healthy blood vessels, and then stiffened using light to mimic vessel scarring in PAH. These 3D-bioprinted models showed that stiffening the microenvironment around female IPAH hPAAFs led to hPAAF activation. On both the protein and gene-expression levels, cellular activation markers significantly increased in stiffened samples and were highest in IPAH patient-derived cells. Treatment with a selective estrogen receptor modulator reduced expression hPAAF activation markers, demonstrating that hPAAF activation is a one pathologic response mediated by estrogen signaling in the vasculature, validating that drugs currently in clinical trials could be evaluated in sex-specific 3D-bioprinted pulmonary artery adventitia models.
肺动脉高压（PAH）对男性和女性患者的影响方式不同。女性患者对疾病的易感性更高（女性与男性的比例为4:1），但诊断后比男性患者寿命更长。这种复杂的性别差异被称为雌激素悖论。先前的研究表明，雌激素信号在肺血管中可能是病理性的，而在心脏中具有保护作用，但PAH中这些性别差异的潜在机制仍不清楚。PAH是一种肺血管疾病，会导致小血管瘢痕形成，导致血流受损和血压升高。随着时间的推移，这种血压升高会对心脏造成损害。以前许多关于PAH的研究依靠雄性细胞或来源未公开的细胞进行建模。在这里，我们展示了一个动态的、3D生物打印模型，该模型整合了来自女性患者的细胞和循环性激素，以专门研究女性患者如何应对微环境硬度和性激素信号的变化。基于聚（乙二醇）-α-甲基丙烯酸酯（PEGαMA）的水凝胶，含有来自特发性PAH（IPAH）或对照供体的女性人肺动脉外膜成纤维细胞（hPAAFs），通过3D生物打印来模拟肺动脉外膜。这些生物材料最初很柔软，就像健康血管一样，然后用光照使其变硬，以模拟PAH中的血管瘢痕形成。这些3D生物打印模型表明，使女性IPAH hPAAFs周围的微环境变硬会导致hPAAF激活。在蛋白质和基因表达水平上，细胞激活标志物在变硬的样本中显著增加，在IPAH患者来源的细胞中最高。用选择性雌激素受体调节剂治疗可降低hPAAF激活标志物的表达，表明hPAAF激活是血管中雌激素信号介导的一种病理反应，证实了目前正在临床试验中的药物可以在性别特异性的3D生物打印肺动脉外膜模型中进行评估。
