Liu Y, Ye T, Sun D, Maynard J, Deisseroth A
Genetic Therapy Program, Sidney Kimmel Cancer Center, San Diego, CA 92121, USA.
Gene Ther. 2006 Aug;13(16):1235-43. doi: 10.1038/sj.gt.3302776. Epub 2006 Apr 13.
We constructed a melanoma-specific oncolytic adenoviral vector Ad.MCDIRESE1.71Hsp3, in which the cytosine deaminase and adenoviral E1A genes linked by the IRES sequence were under the control of a mouse tyrosinase enhancer/promoter transcriptional element in the E1 region of the vector. We also inserted the human heat shock protein 70 (Hsp70) gene driven by the cytomegalovirus promoter into the E3 region of this vector. The RGD-4C peptide was inserted into the HI loop of the fiber knob domain of the Ad.MCDIRESE1.71Hsp3 vector to increase the transduction efficiency of this vector to tumor cells. The Ad.MCDIRESE1.71Hsp3 vector replicates specifically in melanoma cells, and it has a melanoma-specific cytotoxic effect in the presence of 5-fluorocytosine in vitro and in vivo. Moreover, the in vivo killing of tumor cells associated with the overexpression of Hsp70 generated by the intratumoral injection of the Ad.MCDIRESE1.71Hsp3 vector into established subcutaneous tumors can lead to the suppression of tumor growth and potent melanoma-specific systemic immune responses.
我们构建了一种黑色素瘤特异性溶瘤腺病毒载体Ad.MCDIRESE1.71Hsp3,其中通过内部核糖体进入位点(IRES)序列连接的胞嘧啶脱氨酶和腺病毒E1A基因受载体E1区中鼠酪氨酸酶增强子/启动子转录元件的控制。我们还将由巨细胞病毒启动子驱动的人热休克蛋白70(Hsp70)基因插入该载体的E3区。将RGD-4C肽插入Ad.MCDIRESE1.71Hsp3载体纤维钮结构域的HI环中,以提高该载体对肿瘤细胞的转导效率。Ad.MCDIRESE1.71Hsp3载体在黑色素瘤细胞中特异性复制,并且在体外和体内存在5-氟胞嘧啶的情况下具有黑色素瘤特异性细胞毒性作用。此外,通过向已建立的皮下肿瘤内注射Ad.MCDIRESE1.71Hsp3载体产生的与Hsp70过表达相关的肿瘤细胞在体内的杀伤可导致肿瘤生长的抑制和强大的黑色素瘤特异性全身免疫反应。