Ex vivo therapy of PAH utilizing hiPSC derived ECs in a SuNx PAH Rat model and development of an organ specific BMPR2 KO animal model in rat and pig

在 SuNx PAH 大鼠模型中利用 hiPSC 衍生的 EC 进行 PAH 离体治疗，并在大鼠和猪中开发器官特异性 BMPR2 KO 动物模型

• 批准号: 317775478
• 负责人: Professor Dr. Georg Hansmann
• 金额: --
• 依托单位: Klinik für Herz-, Thorax-, Transplantations- und Gefäßchirurgie (HTTG)
• 依托单位国家: 德国
• 项目类别: Clinical Research Units
• 财政年份: 2016
• 资助国家: 德国
• 起止时间: 2015-12-31 至 2020-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/317775478?language=en
• 关键词: Ex; vivo; therapy; PAH; utilizing

Pulmonary arterial hypertension (PAH) is a progressivevasoconstrictive disease. Due to missing curative therapy options, ithas a 5-year mortality up to 50%. Based on the first funding periodone major goal of the current project is to finally establish the new,innovative therapy concept, which bases on the replacement of thedysfunctional pulmonary endothelial cells by (functionalized) hiPSCderived endothelial cells in an ex vivo lung perfusion system (EVLP).This cellular ex vivo therapy will be evaluated on the one hand in thesugen/normoxic PAH rat model, established in the first funding period.As current animal models do not represent the impact of loss offunction mutations in the BMPR2 gene, which can be detected in 70%of the patients with familial PAH, this clinically relevant BMPR2 knockout (KO) PAH rat model will be developed and established to be usedfor evaluation of the cell therapy concept. Therefore, by directtargeting of the pulmonary endothelial cells in the EVLP the BMPR2KO –PAH model in rats and in pigs will be investigated for thegeneration of clinically relevant data. In the EVLP innovative geneediting methods will be combined with non-integrating viral vectors toachieve a targeted KO of the BMPR2 gene in the pulmonaryendothelium. Besides the detailed characterization of the PAHphenotype in the BMPR2-KO model compared to the well-establishedsugen/normoxic model the effect of the ex vivo cell therapy will beevaluated in both animal models. Detailed comparison of treatmenteffects in both animal models will allow to draw inter alia conclusionswith respect to the underlying mechanisms for PAH development. Inaddition, the newly established ex vivo gene editing will be utilized toinduce a BMPR2 KO in the pulmonary endothelial layer of pig lungs togenerate a PAH large animal model to allow for further translationaldevelopment of the established cell therapy concept.

肺动脉高压（PAH）是一种进行性血管狭窄性疾病.由于缺乏治愈性治疗选择，其5年死亡率高达50%。在第一个资助期的基础上，本项目的一个主要目标是最终建立新的、创新的治疗概念，其基础是通过在离体肺灌注系统（EVLP）中的（功能化的）hiPSC衍生的内皮细胞。这种细胞离体疗法将一方面在sugen/常氧PAH大鼠模型中进行评价，由于目前的动物模型不能代表BMPR 2基因功能缺失突变的影响，这可以在70%的家族性PAH患者中检测到，这种临床相关的BMPR 2敲除（KO）PAH大鼠模型将被开发和建立，用于评估细胞治疗概念。因此，通过直接靶向EVLP中的肺内皮细胞，将研究大鼠和猪中的BMPR 2 KO-PAH模型以产生临床相关数据。在EVLP中，创新的基因编辑方法将与非整合病毒载体相结合，以实现肺内皮中BMPR 2基因的靶向KO。除了详细描述BMPR 2-KO模型中PAH表型与已建立的sugen/常氧模型相比的特征外，还将在两种动物模型中评价离体细胞治疗的效果。详细比较两种动物模型的治疗效果，将有助于得出关于PAH发展的潜在机制的结论。此外，新建立的离体基因编辑将用于在猪肺的肺内皮层中诱导BMPR 2 KO，以产生PAH大动物模型，从而允许进一步开发已建立的细胞治疗概念。