AMPK/NADPH协同蛋白磷酸酶4靶向ACC1调控脂肪酸代谢重编程促进肾透明细胞癌转移的分子机制

以脂肪异常沉积为特点的肾透明细胞癌（ccRCC）发病率高、转移发生早、病程进展快。我们发现ccRCC中脂肪酸合成限速酶ACC1异常活化可能参与肿瘤转移，但未见报道。结合研究进展，提出假说：ACC1异常活化介导脂肪酸代谢重编程，影响肿瘤转移；ACC1活性调控依赖于AMPK磷酸化、PP4去磷酸化、底物NADPH变构，以及AMPK与NADPH的协同作用。据此，本课题首先明确ACC1异常活化与ccRCC转移及患者不良预后的关系；进而用ACC1敲低/过表达细胞、裸鼠转移瘤模型和LC-MS等探讨ACC1介导的脂肪酸代谢重编程在肿瘤转移中的作用；最后用AMPK与PP4敲低/过表达细胞及相应激活/抑制剂、或改变NADPH水平，通过Co-IP、免疫荧光等方法阐明ACC1活性调控的分子机制。本研究聚焦ccRCC的转移，从ACC1的活性调控到下游效应与机制展开研究，为ccRCC转移的靶向干预提供线索和依据

Clearn cell Renal cell carcinoma (ccRCC) characterized by abnormal fatty deposition has a high morbidity, early metastasis and rapid progression.In previous studies, we found that abnormal activated ACC1, the rate-limiting enzyme of fatty acid synthesis, might be positively correlated to ccRCC metastasis,but no related report. Combined with recent research progresses, it is hypothesized that abnormal activated ACC1 mediates the fatty acid metabolism reprogramming to affect tumor metastasis, and the regulation of ACC1 activation derived from ACC1 is directly phosphorylated by AMPK, dephosphorylated by PP4, allosteric modulation by the substrate NADPH, and synergistic action by AMPK and NADPH. To test this hypothesis, we will firstly determine the correlation between ACC1 aberrant activation and malignant degree, as well as prognosis in ccRCC by clinical specimens analysis. Then, we want to verify the role of ACC1 mediated fatty fatty acid metabolism reprogramming in tumor metastasis by ACC1 knockdown/overexpression cells, tumor metastatic mode of nude mouse and LC-MS. Finally, through the cell models of knockdown/overexpression of AMPK or PP4, some activator or inhibitor treatment, different NADPH level, we attempt to elucidate the molecular mechanism of ACC1 activity regulation by Co-IP, immunofluorescence and other methods. Taken together, our study is designed to focus on ccRCC metastasis, from the regulation of ACC1 activity to its downstream effect and molecular mechanism, which offers potential clues and evidences for research of metastasis and targeted treatment intervention in ccRCC。