索拉菲尼联合自杀基因调控的异基因CIK细胞治疗肝癌的实验研究

Hepatocellular carcinoma（HCC）is a serious threat to human health, while the overall efficacy of the current treatment is still unsatisfied. Our previously study has reported that allogeneic cytokine-induced killer cells(aCIK) transduced with suicide gene had significant anti-HCC effect, importantly, administration of the suicide gene’s prodrug allows providing a safe and effective prevention of the major complications of aCIK’s alloreactivity, graft versus host disease (GvHD), therefore demonstrated proof-of-concept of using aCIK to treat solid tumors. The key to improve the treatment effect of HCC is how to achieve the HCC targeted of aCIK : we will first study to improve the ability of aCIK hepatic homing in order to increase antitumor effect on HCC. Chemotactic factor 12 (SDF-1) and its specific receptor CXCR4 constitutes of the SDF-1 / CXCR4 axis which can induce lymphocyte directed migration and hepatic metastasis of several kind of tumor cells. Utilizing the high expression level of SDF-1 in hypoxia environment of HCC as well as adjacent cancerous tissue during the course of anti-VEGF effect of sorafenib against HCC, we plan to transduce aCIK cells with CXCR4 in order to increase aCIK homing property to the tumor location of HCC through the navigation function of SDF-1 / CXCR4 axis, thus provide a more stronger anti-HCC effect together with sorafenib. The present project has good research foundation and innovation, could provide a new effective method of adoptive cellular immunotherapy for the treatment of HCC and, finally, represents good perspectives of clinical application.

肝癌是严重威胁人类健康的恶性肿瘤，目前的治疗方式总体疗效仍不满意。我们前期研究首次报道了转染自杀基因的异基因CIK细胞（aCIK）有明显抗HCC的作用，自杀基因前体药物可有效预防异体反应的主要并发症GvHD，揭示了aCIK治疗实体肿瘤的可能性。而提高HCC治疗效果的关键环节是如何实现aCIK的HCC靶向性：我们将首次研究提高aCIK的趋肝性而增加对HCC靶向性杀伤作用。趋化因子12（SDF-1）及其特异受体CXCR4构成的SDF-1/CXCR4轴能诱导淋巴细胞的定向迁移及肿瘤细胞肝转移，利用索拉菲尼抗VEGF作用诱导的HCC及癌旁组织缺氧环境下 SDF-1的高表达，对aCIK细胞转染SDF-1的特异受体CXCR4，通过SDF-1/CXCR4轴的导航作用，实现效应aCIK的趋肝性迁移，协同索拉菲尼发挥更强大的抗HCC作用。本研究具有好的研究基础及创新性，可为肝癌的综合治疗提供新的有效方式。

肝癌是严重威胁人类健康的恶性肿瘤，目前的治疗方式总体疗效仍不满意。我们前期研究首次报道了转染自杀基因的异基因CIK细胞（aCIK）有明显抗HCC的作用，自杀基因前体药物可有效预防异体反应的主要并发症GvHD，揭示了aCIK治疗实体肿瘤的可能性。而提高HCC治疗效果的关键环节是如何实现aCIK的HCC靶向性：本研究拟通过提高aCIK的趋肝性而增加对HCC靶向性杀伤作用。趋化因子12（SDF-1）及其特异受体CXCR4构成的SDF-1/CXCR4轴能诱导淋巴细胞的定向迁移及肿瘤细胞肝转移，利用索拉菲尼抗VEGF作用诱导的HCC及癌旁组织缺氧环境下SDF-1的高表达，对aCIK细胞转染SDF-1的特异受体CXCR4，通过SDF-1/CXCR4轴的导航作用，实现效应aCIK的趋肝性迁移，协同索拉菲尼发挥更强大的抗HCC作用。本研究发现，iCasp9自杀基因系统及CXCR4双质粒转染后aCIK细胞在体外具有明显的细胞毒作用，可发挥明显的HCC细胞杀灭作用，证实了双基因转染并未影响细胞功能，而自杀基因的前体药物CID可以快速诱导aCIK细胞的凋亡，从而预防GVHD的发生。通过门静脉注射Huh-7肝癌细胞株可成功建立HCC原位模型，索拉菲尼可在发挥抗肿瘤的作用时诱导原位HCC模型中肿瘤组织的局部SDF-1高表达，而通过SDF-1/CXCR4轴的趋化作用，成功诱导了表达CXCR4的aCIK细胞的趋肝性，和对照组相比明显增加了aCIK细胞在肝癌组织中的局部浓度，增加了aCIK细胞的抗HCC作用，而aCIK的前体药物CID体内可以快速诱导aCIK细胞的凋亡，从而预防GVHD的发生和发展。通过本课题的研究，为肝癌的治疗提供了一个新的治疗理念，为过继免疫治疗治疗实体肿瘤提供了一种安全、有效的治疗策略，随着过继免疫治疗实体肿瘤靶向性问题的解决，也为我们后续的课题申请及研究奠定了基础。