源自诱导多能干细胞 (iPSC) 的血管类器官 (VO) 有望成为体外疾病模型和药物筛选平台。然而，它们忠实再现人类血管疾病和细胞组成的能力仍不清楚。在这项研究中，我们证明，与非糖尿病 VO (ND-VO) 相比，源自糖尿病供体 iPSC 的 VO (DB-VO) 表现出血管功能受损。 DB-VO 表现出活性氧 (ROS) 水平升高、线粒体含量和活性升高、促炎细胞因子增加以及体内血液灌注恢复减少。通过全面的单细胞 RNA 测序，我们揭示了 DB-VO 内血管细胞类型和簇之间的分子和功能差异以及信号网络。我们的分析确定了 VO 内的主要血管细胞类型（内皮细胞 [EC]、周细胞和血管平滑肌细胞），突出了 EC 和壁细胞之间的二分法。我们还证明了使用器官特异性分化因子进行额外诱导的潜在需要，以促进 VO 中的器官特异性身份。此外，我们观察到 VO 内的基础异质性以及 DB-VO 和 ND-VO 之间的显着差异。值得注意的是，我们确定了 DB-VO 特异的 EC 亚群，在 ROS 通路中表现出过度表达，而在血管生成标志中表现不足，表明糖尿病中异常血管生成的迹象。我们的研究结果强调了 VO 在糖尿病血管病变建模中的潜力，强调了研究 VO 内细胞异质性对于疾病建模和药物发现的重要性，并提供了 EC 中 GAP43（神经调节蛋白）表达的证据，特别是在 DB-VO 中，这对血管疾病具有重要意义。发育和疾病。© 作者 2024。由牛津大学出版社出版。

Vascular organoids (VOs), derived from induced pluripotent stem cells (iPSCs), hold promise as in vitro disease models and drug screening platforms. However, their ability to faithfully recapitulate human vascular disease and cellular composition remains unclear. In this study, we demonstrate that VOs derived from iPSCs of donors with diabetes (DB-VOs) exhibit impaired vascular function compared to non-diabetic VOs (ND-VOs). DB-VOs display elevated levels of reactive oxygen species (ROS), heightened mitochondrial content and activity, increased proinflammatory cytokines, and reduced blood perfusion recovery in vivo. Through comprehensive single-cell RNA sequencing, we uncover molecular and functional differences, as well as signaling networks, between vascular cell types and clusters within DB-VOs. Our analysis identifies major vascular cell types (endothelial cells [ECs], pericytes, and vascular smooth muscle cells) within VOs, highlighting the dichotomy between ECs and mural cells. We also demonstrate the potential need for additional inductions using organ-specific differentiation factors to promote organ-specific identity in VOs. Furthermore, we observe basal heterogeneity within VOs and significant differences between DB-VOs and ND-VOs. Notably, we identify a subpopulation of ECs specific to DB-VOs, showing overrepresentation in the ROS pathway and underrepresentation in the angiogenesis hallmark, indicating signs of aberrant angiogenesis in diabetes. Our findings underscore the potential of VOs for modeling diabetic vasculopathy, emphasize the importance of investigating cellular heterogeneity within VOs for disease modeling and drug discovery, and provide evidence of GAP43 (neuromodulin) expression in ECs, particularly in DB-VOs, with implications for vascular development and disease.© The Author(s) 2024. Published by Oxford University Press.