TMTP1修饰的第三代腺病毒载体介导tBid表达对卵巢癌治疗效应的研究

Epithelial ovarian cancer is the commonest cause of gynaecological cancer-associated death，most women with advanced disease will develop many episodes of recurrent disease with progressively shorter disease-free intervals. These episodes culminate in chemoresistance and ultimately bowel obstruction. In order to improve chemotherapy resistance of recurrent ovarian cancer, the present study will screen a novel conditionally-replicative adenovirus. The strategy of genetic engineering of adenovirus consists of the incorporation of TMTP1(NVVRQ) peptide into HVR2，HVR5 or HVR7 on Hexon protein, deletion of 920-946 bp on E1A, and incorporation of luciferase as a reporter gene or tBid as a therapeutic gene on E3 protein. Then, we will verify the Tumor targeting, Hepatic un-targeting of above adenovirus vector and demonstrate anti-tumor effect of a therapeutic regimen of DDP with the screening novel adenovirus through constructing the cell model and animal model in vitro and in vivo.

卵巢癌腹腔转移和铂类耐药是患者死亡的首要原因。本实验室前期筛选的高转移肿瘤靶向肽TMTP1为卵巢癌的治疗提供了新的思路。本项目将基于第三代腺病毒载体（Ad5/dE1A/ADP），将TMTP1的DNA序列分别嵌入第三代腺病毒HVR2、HVR5或HVR7中，并使用荧光素酶作为报告基因插入Ad5-E3的ADP（adenovirus death protein）区域，获得4种新型腺病毒载体，比较并筛选出一种具有安全性好、肿瘤感染效率高、免疫原性低为特性的TMTP1修饰的条件复制性腺病毒载体，以用于卵巢癌的基因治疗的载体；然后将凋亡基因tBid重组入筛选出的腺病毒载体ADP区域，体内体外实验验证其单独或联合顺铂对卵巢癌的杀伤效应和体内安全性。 寄希望获得一种新型腺病毒载体Ms-tBid，其联合铂类药物可改善对铂类耐药型卵巢癌的治疗效果。

卵巢癌腹腔转移和铂类耐药是患者死亡的首要原因。本实验室前期筛选的高转移肿瘤靶向肽TMTP1为卵巢癌的治疗提供了新的思路。本项目将基于第三代腺病毒载体（Ad5/dE1A/ADP），将TMTP1的DNA序列分别嵌入第三代腺病毒HVR5中，体外实验显示M0-TMTP1对高转移卵巢癌细胞系转染效率大大增加，在卵巢癌腹腔转移模型中证实M0-TMTP1在腹腔转移灶中病毒载量增加,全身分布实验显示其内肝脏内摄取显著降低，几乎检测不到，因此体内体外实验均证实TMTP1插入HVR5区赋予条件复制性腺病毒载体新的肿瘤靶向性；然后将凋亡基因tBid重组入腺病毒载体ADP区域，体内体外实验验证其具备单独或联合顺铂对耐药卵巢癌肿瘤的杀伤效应和体内安全性。结合M0-TMTP1对肿瘤的靶向性和M0-tBid对铂类耐药的杀伤效应本研究获得M11，体外实验验证了腺病毒载体对肿瘤细胞的嵌合靶向性和tBid的逆转铂类耐药特性；该项工作发表两篇SCI论文（均发表在Human Gene Therapy，IF 5.695），具体见附件！本研究证实TMTP1嵌合入HVR5增加肿瘤靶向性的可行性和客服了溶瘤腺病毒在卵巢癌应用的局限性。该研究成果已进入临床转化阶段（IND前）。