Adenoviral-mediated imaging of gene transfer using a somatostatin receptor-cytosine deaminase fusion protein.
Adenoviral-mediated imaging of gene transfer using a somatostatin receptor-cytosine deaminase fusion protein.
复制标题
使用生长抑素受体-胞嘧啶脱氨酶融合蛋白进行腺病毒介导的基因转移成像。
DOI：
10.1038/cgt.2015.14
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发表时间：
2015
影响因子：
6.4
通讯作者：
Rogers,BE
中科院分区：
文献类型：
--
作者：
Lears,KA;Parry,JJ;Andrews,R;Nguyen,K;Wadas,TJ;Rogers,BE
自杀基因疗法是一种过程，通过这种过程，细胞被给予一种基因，该基因编码一种能够将无毒前药转化为活性毒素的蛋白质。胞嘧啶脱氨酶（CD）由于能够将前药5-氟胞嘧啶（5-FC）转化为毒性化合物5-氟尿嘧啶（5-FU）而被广泛研究作为自杀基因治疗的手段。然而，基因转移的程度是预测治疗结果的限制因素。非侵入性地监测基因转移的能力将加强治疗的效率。在这方面，我们已经构建和评估了一个复制缺陷型腺病毒（Ad）含有人生长抑素受体亚型2（SSTR 2）融合的C-末端酵母CD基因的非侵入性监测的基因转移和治疗。在乳腺癌细胞中体外评价所得Ad（AdSSTR 2-yCD）以确定融合蛋白的功能。这些研究表明，SSTR 2和yCD在结合试验、转化试验和细胞毒性试验中均具有功能。体内研究使用转化测定、生物分布研究和小动物正电子发射断层扫描（PET）成像研究类似地证明了功能性。总之，融合蛋白已被验证为可用于yCD表达的非侵入性成像，并将在未来进行评估，以监测基于yCD的治疗。
Suicide gene therapy is a process by which cells are administered a gene that encodes a protein capable of converting a nontoxic prodrug into an active toxin. Cytosine deaminase (CD) has been widely investigated as a means of suicide gene therapy owing to the enzyme’s ability to convert the prodrug 5-fluorocytosine (5-FC) into the toxic compound 5-fluorouracil (5-FU). However, the extent of gene transfer is a limiting factor in predicting therapeutic outcome. The ability to monitor gene transfer, non-invasively, would strengthen the efficiency of therapy. In this regard, we have constructed and evaluated a replication-deficient adenovirus (Ad) containing the human somatostatin receptor subtype 2 (SSTR2) fused with a C-terminal yeast CD gene for the non-invasive monitoring of gene transfer and therapy. The resulting Ad (AdSSTR2-yCD) was evaluated in vitro in breast cancer cells to determine the function of the fusion protein. These studies demonstrated that both the SSTR2 and yCD were functional in binding assays, conversion assays and cytotoxicity assays. In vivo studies similarly demonstrated the functionality using conversion assays, biodistribution studies and small animal positron-emission tomography (PET) imaging studies. In conclusion, the fusion protein has been validated as useful for the non-invasive imaging of yCD expression and will be evaluated in the future for monitoring yCD-based therapy.