ATF2-miRNA9在结节性硬化症发生发展中的作用及机制研究

Tuberous sclerosis complex (TSC) is an autosomal dominant multisystem neurocutaneous syndrome characterized by the development of multiple hamartomas distributed throughout the body. About 80% of adult TSC patients have multiple and bilateral renal angiomyolipomas (TSC-RAML), which is the most common cause of TSC-associated death. The dual mRNA/miRNA expression profiling were performed in the TSC-RAML and control tissue, we found activating transcription factor 2 (ATF2) and miRNA9 co-highly-expression in TSC-RAML tissue. Pre-experiment found suppression of miRNA-9 expression promotes TSC cell apoptosis and inhibits TSC cell cycle transition in TSC cell model. Bioinformatics analysis reveals that there are multiple binding sites ATF2 in the upstream of miRNA-9 gene. We also found that the downstream multiple targets of miRNA-9 with potential function to promote cell cycle transition and inhibit apoptosis. Therefore, we propose the scientific hypotheses that high expression of ATF2-miRNA9 could promote the initiation and development of tuberous sclerosis complex. This project aims to analysis and build TSC-RAML tissue miRNA/mRNA regulatory networks from the system level, and investigate the molecular mechanisms of activating “ATF2-miRNA9” signaling for promoting promote pathogenesis TSC-RAML occurrence and progression. Our goal is to provide new molecular target for the prevention and therapy of TSC-RAML.

结节性硬化症（TSC）是一种罕见的累及多器官系统的常染色体显性遗传疾病，累及肾脏表现为多发血管平滑肌脂肪瘤（TSC-RAML）可见于约80%的患者，是成年患者的首位致死原因。课题组应用mRNA/miRNA双重表达谱检测发现，激活转录因子2（ATF2）和miRNA9在TSC-RAML组织中共同高表达，预实验发现在TSC细胞模型中下调miRNA9可抑制细胞周期转换及促进细胞凋亡。生物信息学分析发现miRNA9基因上游存在ATF2结合位点，miRNA9下游潜在多个靶点与细胞周期、凋亡密切相关。因此，我们提出高表达ATF2-miRNA9可促进TSC-RAML发生发展的科学假说。本项目拟从系统水平上解析和构建TSC-RAML的miRNA/mRNA调控网络，深入研究ATF2-miRNA9促进TSC-RAML发生发展的作用及机制，为防治该病提供新的分子靶点与理论依据。

结节性硬化症（TSC）是一种罕见的累及多器官系统的常染色体显性遗传疾病，累及肾脏表现为多发血管平滑肌脂肪瘤（TSC-RAML）,可见于约80%的患者,是成年患者的首位致死原因。在课题组前期研究中，mRNA/miRNA双重表达谱检测的结果提示，激活转录因子2（ATF2）和miRNA9在TSC-RAML组织中共同高表达，预实验发现在TSC细胞模型中下调miRNA9可抑制细胞周期转换及促进细胞凋亡。生物信息学分析发现miRNA9基因上游存在ATF2结合位点。因此，我们提出高表达ATF2-miRNA9可促进TSC-RAML发生发展的科学假说。.本项目实施过程中发现：ATF2和miR-9的表达在TSC患者AML组织、TSC2-/-细胞株中均显著升高。该通路可抑制BCL2L11和PTEN的表达，从而促进抑制凋亡，促进细胞增殖，阻断ATF2-miR-9可抑TSC2-/-细胞的活率，抑制裸鼠成瘤实验中的肿瘤大小；过表达miR-9则可通过抑制BCL2L11和PTEN，从而促进TSC2-/-细胞的增殖，促进肿瘤发展。.本项目研究成果显示，TSC-RAML中存在ATF2-miR-9-BCL2L11/PTEN的信号通路，导致TSC2缺陷的细胞中凋亡水平下架，自噬水平升高，从而促进细胞的增殖。ATF2可能作为TSC靶向治疗的靶点，而凋亡、自噬通路的靶向治疗也有望成为结节性硬化新的治疗手段。