自噬相关转录因子上调细胞自噬促进肝癌转移的机制及基于自噬转录调节的肝癌组织特异性靶向治疗研究

Hepatocellular carcinoma (HCC) is one of the most fatal cancers in China. Metastasis is responsible for the poor prognosis of HCC and it has been linked to cell death resistance. Autophagy is an important survival mechanism under conditions of cell stress. In our preview studies, we have demonstrated that autophagy played an important role in HCC metastasis due to autophagy-mediated facilitation of lung colonization of HCC cells. Autophagy inhibition can significantly decrease the incidence of pulmonary metastases. Recently we found that autophagy was consistly upregulated in metastatic foci through transcriptional mechanism and targeting autophagy-associated transcriptional fatctors can significantly suppressed lung metastsis of HCC. The purpose of this study aims to clarify the molecular mechanism of the upregulation of autophagy of HCC metastasis and develop more effective autophagy-associated HCC tissue-specific multiple-genes target therapy. Firslty, we verified the role of transcriptional regulation in the consistant activation of autohpagy in lung metastasis. Then high-throughout detction of transcriptional factors will be performed and the autophagy-associated transcriptional factors will be determined. The potential value of autophagy-associated transcriptional factors for treatment of lung metastassis will be evaluated. We will performed the ChiP-seq analysis to clarify the interplay between the autophagy-associated transcriptional factors and autophagy genes. A new HCC tissue-specific multiple-genes target therapy system AFP-Cas9-gRNA will be constructed. Based on the results from molecular mechanism investigations, highly efficient autophagy- associated transcriptional regualtion based HCC tissue-specific target therapy will be design and evaluted in vivo.

肝细胞癌是严重威胁我国人民健康的恶性疾病。转移复发研究是关键。细胞自噬是维持细胞生存的重要机制。前期研究证实细胞自噬在肝癌转移过程中起重要作用，自噬在肝癌转移过程中激活上调，促进肝癌细胞定植转移，抑制自噬可抑制肝癌肺转移发生。近期我们发现转录调节是肝癌细胞肺转移过程中自噬上调的主要机制，并且靶向STAT3等自噬相关转录因子对肝癌肺转移的治疗效果远远超过靶向自噬基因，具有重大研究价值。本研究拟探讨自噬在肝癌肺转移过程中激活上调的机制并建立基于自噬调节机制的靶向治疗策略。首先进一步明确转录调节在肝癌肺转移自噬上调中的作用及相关转录因子，继而通过ChIP-seq等分析明确其上调自噬基因表达的作用机制，并进一步明确其在肝癌肺转移治疗中的潜在价值。同时，构建新的肝癌组织特异性多基因作用系统AFP-Cas9-gRNAs，并在自噬转录调节机制研究基础上构建基于自噬调节机制的高效靶向治疗策略。

肝细胞癌是严重威胁我国人民健康的恶性疾病。转移复发研究是关键。细胞自噬是维持细 胞生存的重要机制。前期研究证实细胞自噬在肝癌转移过程中 起重要作用，自噬在肝癌转移过程中激活上调，促进肝癌细胞定植转移，抑制自噬可抑制肝癌肺转移发生。转录调节被发现是肝癌细胞肺转移过程中自 噬上调的主要机制，并且靶向自噬相关转录因子对肝癌肺转移的治疗效果远远超过靶向自噬基因，具有重大研究价值。本研究探讨自噬在肝癌肺转移过 程中激活上调的机制并尝试在前期研究基础上建立基于自噬调节机制的靶向治疗策略。本研究首先进一步通过体内研究明确了转录调节在肝癌肺转移自 噬上调中的作用，进一步明确自噬上调的相关转录因子，发现STAT3、TFEB、CREB等因子在其中起重要作用，并探讨了其上调自噬基因表达的作用机制 ，发现自噬调节相关转录因子的活化机制与肿瘤微环境有关。构建了以AAV-DJ8为载体的新的肝癌组织特异性多基因作用系统AFP-Cas9-gRNAs，并于体内 验证基于自噬转录因子的靶向治疗效果。本研究部分阐明自噬在肝癌肺转移过程中的作用机制，明确了自噬相关的转录调节因子，获得新的作用靶标， 研究形成新的更好的肝癌组织特异性靶向治疗系统，并一定程度证实其潜在价值。