PIEZO1基因介导AKT/eNOS信号通路在肺血栓栓塞症中的作用机制研究

肺血栓栓塞症（PTE）是一种高致死性疾病，严重威胁人类生命健康。其发生与遗传因素相关，但确切的易感机制尚不完全明确。尽管已有部分基因被证实与PTE相关，但仍有大量易感基因尚未被发现，尤其是针对内皮功能、炎症等作用机制的基因。我们前期通过对家系测序分析发现，PIEZO1基因参与PTE的发生，但致病机制尚不清楚。Piezo1通过下游AKT/eNOS信号通路及维持Ca2+浓度，维持保证内皮细胞（EC）功能稳态。若EC功能损伤，PTE发生风险将显著增加。基于此，我们提出假说，PIEZO1基因突变导致Piezo1表达及功能改变，通过对AKT/eNOS信号通路以及Ca2+浓度影响，降低eNOS活性，引起EC功能损伤，参与PTE的发生。本研究拟从细胞、动物水平，应用CRISPR/Cas9等技术，探讨PIEZO1基因突变参与PTE发生的具体机制，为PTE精准防治提供新思路。

Pulmonary thromboembolism (PTE) is a life-threatening disease. Genetic factors are involved in the etiology of PTE but the mechanisms underlying the genetic susceptibility are unclear. Although some of genes have been confirmed to be related to PTE, a large number of susceptibility genes have not been discovered, especially the genes are known for their roles in endothelial function and inflammation. Our preliminary pedigree study has found that mutations of the PIEZO1 gene may be involved in the development of PTE. However, the exact pathogenic mechanism is unexplained. As a mechanosensitive channel protein, Piezo1 maintains endothelial cell (EC) homeostasis through the regulation of intracellular Ca2+ concentration, and the protein kinase B/endothelial nitric oxide synthase (AKT/eNOS) signaling pathway. Changes in eNOS activity may cause EC dysfunction and increase risk of developing PTE. Therefore, we hypothesize that PIEZO1 mutations reduce eNOS activity by altering inactivating the AKT/eNOS signaling pathway and the intracellular Ca2+ concentration, resulting in EC damage that may contribute to the development of PTE. This study intends to use the CRISPR/Cas9 gene editing method to assess the effects PIEZO1 mutations in vitro and in vivo. Our study will explore the specific mechanisms by which PIEZO1 mutations could cause PTE, and provide new insight on prevention and early detection of PTE.