In vitro- and in vivo-targeted tumor lysis by an MMP2 cleavable melittin-LAP fusion protein
In vitro- and in vivo-targeted tumor lysis by an MMP2 cleavable melittin-LAP fusion protein
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DOI：
10.3892/ijo_00000396
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发表时间：
2009-10-01
影响因子：
5.2
通讯作者：
Yu, Xianzhong
中科院分区：
文献类型：
--
作者：
Holle, Lori;Song, Wen;Yu, Xianzhong
化疗是癌症的主要治疗选择之一，但由于化疗药物的全身毒性，其有效性受到严重限制。因此，在肿瘤治疗中需要更有针对性的方法是显而易见的。肿瘤活化剂将降低全身毒性以及增加治疗的功效。先前已经表明，pro-TGF-β的潜伏期是由形成保护屏障的两个潜伏期相关肽（LTP）的二聚化赋予的，所述保护屏障在被基质金属蛋白酶（MMP）激活时被切割掉。还已经显示，这种β肽与其他细胞因子的融合可以赋予它们的潜伏期。在本研究中，制备了具有编码肿瘤活化的促细胞溶解肽的融合基因的重组腺病毒，其中TGF-β的β-D结构域与蜂毒肽（一种有效的细胞溶解毒素）融合，在两者之间具有MMP 2切割位点。体外研究表明，蜂毒肽-MMP 2-β重组腺病毒可被MMP 2激活，释放出游离的蜂毒肽，裂解靶细胞。体内研究显示，与对照小鼠相比，蜂毒肽-MMP 2-β重组腺病毒处理的小鼠中B16肿瘤体积减少约70%。在处理的小鼠中未观察到显著的全身毒性。
Chemotherapy is one of the main treatment options for cancer, but the effectiveness of chemotherapeutic drugs is severely limited due to their systemic toxicity. Therefore, the need for a more targeted approach in tumor treatment is obvious. A tumor-activated agent would decrease systemic toxicity as well as increase the efficacy of the treatment. It has previously been shown that the latency of pro-TGF-beta is conferred by dimerization of two latency-associated peptides (LAP) that form a protective shield, which is cleaved off upon activation by matrix metalloproteinases (MMPs). It has also been shown that the fusion of this LAP peptide with other cytokines can confer their latency. In the present study, a recombinant adenovirus with a fusion gene encoding a tumor-activated pro-cytolytic peptide was made in which the LAP domain of TGF-beta was fused with melittin, a potent cytolytic toxin, with an MMP2 cleavage site in between the two. In vitro studies show that the melittin-MMP2-LAP recombinant adenovirus can be activated by MMP2 which leads to the release of free melittin to lyse the target cells. In vivo studies show approximately a 70% decrease in B16 tumor volume in melittin-MMP2-LAP recombinant adenovirus-treated mice as compared to control mice. No significant systemic toxicity was observed in the treated mice.