Mueller Mikala C, Blomberg Rachel, Tanneberger Alicia E, Davis-Hall Duncan, Neeves Keith B, Magin Chelsea M
Department of Bioengineering, University of Colorado, Denver|Anschutz Medical Campus, Aurora 80045, Colorado, United States.
Department of Pediatrics, University of Colorado, Anschutz Medical Campus, Aurora 80045, Colorado, United States.
ACS Biomater Sci Eng. 2025 May 12;11(5):2935-2945. doi: 10.1021/acsbiomaterials.5c00123. Epub 2025 Apr 26.
Pulmonary arterial hypertension (PAH) is a form of pulmonary vascular disease characterized by scarring of the small blood vessels that results in reduced blood flow and increased blood pressure in the lungs. Over time, this increase in blood pressure causes damage to the heart. Idiopathic (IPAH) impacts male and female patients differently, with female patients showing a higher disease susceptibility (4:1 female-to-male ratio) but experiencing longer survival rates postdiagnosis compared to male patients. This complex sex 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 IPAH remain unclear. Many previous studies of PAH relied on male cells or cells of undisclosed origin for modeling. Here, we present a dynamic, three-dimensional (3D)-bioprinted model incorporating cells and circulating sex hormones from female patients to specifically study how female patients respond to changes in microenvironmental stiffness and sex hormone signaling on the cellular level. Poly(ethylene glycol)-α methacrylate (PEGαMA)-based hydrogels containing female human pulmonary artery adventitia fibroblasts (hPAAFs) from 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, which is currently in clinical trials for IPAH treatment, reduced the expression of hPAAF activation markers, demonstrating that hPAAF activation is one pathologic response mediated by estrogen signaling in the vasculature. These results showed the utility of sex-specific, 3D-bioprinted pulmonary artery adventitia models for preclinical drug discovery and validation.
肺动脉高压（PAH）是一种肺血管疾病，其特征是小血管瘢痕形成，导致肺血流量减少和血压升高。随着时间的推移，这种血压升高会对心脏造成损害。特发性肺动脉高压（IPAH）对男性和女性患者的影响不同，女性患者表现出更高的疾病易感性（女性与男性的比例为4:1），但与男性患者相比，诊断后的生存率更长。这种复杂的性别二态性被称为雌激素悖论。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IPAH中这些性别差异的潜在机制仍不清楚。以前许多关于PAH的研究依赖于雄性细胞或来源未公开的细胞进行建模。在这里，我们展示了一种动态的三维（3D）生物打印模型，该模型包含来自女性患者的细胞和循环性激素，以具体研究女性患者在细胞水平上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，就像健康的血管一样，然后通过光照变硬，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，并且在IPAH患者来源的细胞中最高。用一种选择性雌激素受体调节剂进行治疗（目前正在进行IPAH治疗的临床试验），降低了hPAAF激活标志物的表达，表明hPAAF激活是雌激素信号在血管系统中介导的一种病理反应。这些结果表明了性别特异性的、3D生物打印的肺动脉外膜模型在临床前药物发现和验证中的实用性。
