AEP调节M2巨噬细胞自噬过程机理的研究

Asparagine endopeptidase (AEP) has been found to be closely related with innate immunity and clinical pathological changes of tumor. Studies have shown that AEP plays an important role in tumorigenesis, tumor growth and metastasis. However, the molecular mechanism of AEP still remains elusive. We found in previous experiments that AEP deficiency in M2 macrophages impaired the autophagic-lysosomal pathway. Impaired autophagic-lysosomal pathway in AEP-/- M2 macrophages caused pro-inflammatory factor NF-κB overactivation, which led to higher levels of TNFα production. We established spontaneous lung metastasis model and experimental lung metastasis model using murine non-small cell lung cancer cell line D121, and found that AEP KO mice developed more serious lung metastasis than WT mice. The project intends to use Wildtype and AEP KO M2 macrophages differentiated in vitro to analysis the function of AEP on autophagy and inflammation, and how AEP regulates autophagy pathway by PI3K / mTOR pathway. Our researches will clarify that AEP is a critical autophagy modulator in M2 macrophage-involved inflammation and tumor metastasis. We will use AEP conditional-knock mice to screen potential inhibitors of inflammation and tumor.

天冬酰胺内肽酶（Asparagine endopeptidase，AEP）已被证明在先天免疫、肿瘤发生和转移过程中发挥着重要作用，但其中的分子机理仍鲜为人知。我们在前期实验中发现AEP缺失M2巨噬细胞自噬-溶酶体途径缺陷，促炎性通路NF-κB激活增强，炎症因子 TNFα分泌增多。用鼠源非小细胞肺癌细胞系 D121 建立自发性肺转移和实验性肺转移模型，观察到 AEP缺失小鼠 D121 细胞肺转移明显增强。本项目拟利用体外分化Wild Type和AEP缺失的M2巨噬细胞分析AEP对自噬和炎症的影响，深入研究AEP是如何通过PI3K/mTOR通路调节自噬途径，系统地阐明AEP作为自噬调节因子在 M2 巨噬细胞介导的炎症反应和肿瘤转移过程中具有重要作用。同时利用AEP条件敲除鼠分析筛选可能的炎症和肿瘤抑制剂。

天冬酰胺内肽酶（Asparagine endopeptidase，AEP）已被证明在先天免疫、肿瘤发生和转移过程中发挥着重要作用，但相关机理仍有待研究。我们研究发现AEP缺失M2巨噬细胞自噬-溶酶体途径缺陷，促炎性通路NF-κB激活增强；利用体外分化的WT和AEP-/- M2巨噬细胞分析发现AEP调控PI3K/mTOR通路从而调节自噬途径的机理，即AEP通过剪切亚基p85调节PI3K活性，影响PIP2和PIP3转化，从而调控自噬流，影响炎症反应。利用构建的自发性肺转移和实验性肺转移模型，观察到 AEP缺失小鼠肺转移明显增强，其中AEP-/-巨噬细胞具有重要调控作用。研究阐明AEP作为自噬调节因子参与在 M2 巨噬细胞介导的炎症反应和肿瘤转移过程。同时分析筛选自噬调节剂在肿瘤转移中的作用。