Targeting RIG-I with 5'ppp-modified RNA for immunotherapy of hepatocellular carcinoma (HCC)
Targeting RIG-I with 5'ppp-modified RNA for immunotherapy of hepatocellular carcinoma (HCC)
复制标题
使用 5ppp 修饰的 RNA 靶向 RIG-I 用于肝细胞癌 (HCC) 的免疫治疗
DOI：
10.1055/s-0034-1386661
复制
发表时间：
2014
影响因子：
1.3
通讯作者：
Schnurr M.
中科院分区：
文献类型：
--
作者：
Posselt L;Lazic I;Boehmer D;Funk A;Kirchleitner S;Hoffmann S;Adunka T;Endres S;Düwell P;Rothenfusser S;Schnurr M.
背景：肝细胞癌是最常见的原发性肝癌。晚期肿瘤患者的中位生存期不到一年。迫切需要新的治疗选择。胞质解旋酶视黄酸诱导基因I（RIG-I）是病毒5 '-三磷酸-RNA（ppp-RNA）的免疫受体，其激活通过I型IFN和促炎细胞因子导致先天性和适应性免疫。此外，它还促进肿瘤细胞凋亡的内在途径。方法：通过体外转录合成RIG-I的5 'ppp-RNA，并用于分析RIG-I在人（Hep 3B）和鼠（Hepa 1 -6）肝癌细胞中的功能表达。在HCC的原位Hepa 1 -6小鼠模型中体内评价治疗功效。肿瘤诱导七天后，用与聚乙烯亚胺复合的5 'ppp-RNA静脉内治疗小鼠。24小时后处死小鼠，取出肿瘤并测量大小。通过TUNEL染色评估肿瘤细胞死亡，通过qRT-PCR评估细胞因子表达。结果：用ppp-RNA处理人和鼠HCC细胞诱导IRF 3的磷酸化，产生IFN-β和IFN诱导的基因，如CXCL 10，指示完整的RIG-I信号传导。此外，RIG-I激活导致了严重的肿瘤细胞死亡。与对照RNA相比，用ppp-RNA对原位HCC小鼠进行单次治疗显著减小了肿瘤大小，诱导肿瘤组织中I型IFN的产生并导致大量肿瘤细胞死亡。重复ppp-RNA注射治疗小鼠显着延长survival.Conclusion：我们可以证明，RIG-I通路在HCC细胞中是功能性的。RIG-I激活导致I型IFN产生和大量肿瘤细胞在体外和体内死亡。基于RIG-I的免疫疗法为HCC治疗带来了希望，值得进一步评估。
Background: HCC is the most common primary malignant type of liver cancer. Median survival for patients with advanced tumors is less than one year. Novel treatment options are urgently needed. The cytosolic helicase retinoic acid-inducible gene I (RIG-I) is an immune receptor for viral 5'-triphosphat-RNA (ppp-RNA) and its activation leads to innate and adaptive immunity via type I IFN and proinflammatory cytokines. In addition, it promotes the intrinsic pathway of apoptosis in tumor cells. This project focuses on the development of 5'ppp-RNAs targeting RIG-I for HCC therapy.Methods: A synthetic 5'ppp-RNA was generated via in vitro transcription and used to analyze functional RIG-I expression in human (Hep3B) and murine (Hepa1–6) HCC cells. Therapeutic efficacy was evaluated in vivo in the orthotopic Hepa1–6 mouse model of HCC. Seven days after tumor induction mice were treated intravenously with 5'ppp-RNA complexed to polyethylenimine. 24h later mice were sacrificed, tumors removed and size measured. Tumor cell death was assessed by TUNNEL staining and cytokine expression by qRT-PCR. In another experiment survival of RNA-treated mice was monitored.Results: Treatment of human and murine HCC cells with ppp-RNA induced phosphorylation of IRF3, production of IFN-β and IFN-induced genes, such as CXCL10, indicative of intact RIG-I signaling. In addition, RIG-I activation led to profound tumor cell death. A single treatment of mice with orthotopic HCC with ppp-RNA significantly reduced tumor size in comparison to control RNA, induced type I IFN production in the tumor tissue and led to massive tumor cell death. Treatment of mice with repeated ppp-RNA injections significantly prolonged survival.Conclusion: We could demonstrate that the RIG-I pathway is functional in HCC cells. RIG-I activation leads to type I IFN production and massive tumor cell death both in vitro and in vivo. RIG-I-based immunotherapy holds promise for HCC therapy deserving further evaluation.