A-RAF激酶在PARP-1介导的细胞凋亡中的作用机理研究

随着我国人口老龄化加剧，神经退行性疾病的患病率也逐渐升高。PARP-1介导的细胞凋亡(parthanatos)是细胞死亡的重要形式之一，其在神经退行性疾病的发生发展过程中发挥重要作用。目前研究表明PARP-1间接通过AIF导致细胞死亡，但对此具体细胞死亡过程中所涉及的其它蛋白分子尚不清楚。A-RAF是RAF激酶家族的三个重要成员之一。我们前期研究结果显示，A-RAF与PARP-1和AIF均高度结合，且A-RAF可显著促进PARP-1介导的细胞凋亡。本研究拟通过调控A-RAF表达水平，来观察其对PARP-1和AIF功能的影响，探讨A-RAF在PARP-1介导的细胞凋亡中的具体作用机制，同时利用Crispr/Cas9技术在基因组层面筛选parthanatos死亡通路中的其它关键调控蛋白。通过该研究阐明A-RAF在parthanatos通路中的作用机理，为神经退行性疾病的诊断和治疗开辟新的思路。

The prevalence of neurodegenerative diseases is increasing rapidly with the growing trend of our aging population. PARP-1-mediated apoptosis (parthanatos) is one of the major types of cell death, and it is evident that parthanatos plays a significant role in the occurrence and development of various diseases, particularly neurodegenerative diseases. Cumulative research has demonstrated that PARP-1 indirectly causes chromatin condensation and DNA breaks in a AIF-dependent manner, finally resulting in cell death. However, the contributions of other proteins to parthanatos are unclear or have a considerable controversy. RAF kinases are one of key components in the major signaling pathway RAS-RAF-MEK-ERK, and A-RAF is one of three members of RAF family kinase. Our preliminary results demonstrated that A-RAF highly bound to both PARP-1 and AIF. Through A-RAF over-expression or knockout, we found that A-RAF could significantly promote PARP-1-mediated apoptosis. In this project, we will observe the effects of A-RAF on PARP-1 and AIF functions, and explore the mechanism by which A-RAF kinase regulates PARP-1-mediated apoptosis through controlling A-RAF expression levels. We will also identify other key regulatory proteins involved in parthanatos by using Crispr/Cas9 for genome-scale screens. This work will demonstrate the role of A-RAF in PARP-1 mediated apoptosis, clarify the mechanism of parthanatos. Ultimately， this study will give new insight into the diagnosis and treatment of parthanatos-related diseases, particularly neurodegenerative diseases.