Specific membrane receptor gene expression targeted with radiolabeled peptide employing the erbB-2 and DF3 promoter elements in adenoviral vectors
Specific membrane receptor gene expression targeted with radiolabeled peptide employing the erbB-2 and DF3 promoter elements in adenoviral vectors
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DOI：
10.1038/sj.cgt.7700049
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发表时间：
1999-05-01
影响因子：
6.4
通讯作者：
Curiel, DT
中科院分区：
文献类型：
--
作者：
Stackhouse, MA;Buchsbaum, DJ;Curiel, DT
放射免疫疗法受到多种因素的限制，包括单克隆抗体的肿瘤渗透性差和瘤内抗原表达水平低。为了解决这些局限性，设计了一种基因治疗策略，通过基因诱导肿瘤细胞表达增强水平的对放射性标记肽具有高亲和力的膜受体。我们将这种方法指定为遗传放射性同位素靶向策略。为此，使用编码鼠胃泌素释放肽受体（GRPr）的腺病毒载体（AdCMVGRPr）来实现放射性标记铃蟾肽（BBN）的高水平结合。为了实现膜 GRPr 特异性针对肿瘤细胞的遗传诱导，我们构建了两种在肿瘤特异性调控元件 DF3 (AdDF3GRPr) 或 erbB-2 (AderbGRPr) 控制下编码 GRPr 基因的腺病毒载体。我们在一组人乳腺癌、胰腺癌和胆管癌肿瘤细胞系中研究了 AdDF3GRPr 和 AderbGRPr 感染后 [I-125]BBN 与 GRPr 的结合，在所有测试的细胞系中，[I-125]BBN 结合和 GRPr 表达随着 AdCMVGRPr 感染复数的增加而增加。使用 AderbGRPr，表达 erbB-2 的乳腺癌细胞系显示出显着的 GRPr 表达。通过免疫组织化学检测，在感染表达 MUC1 的 AdDF3GRPr 的乳腺癌和胆管癌细胞中观察到类似的结果，但在测试的胰腺细胞系中未观察到类似的结果。因此，具有组织特异性启动子元件的腺病毒载体可用于实现可以用放射性标记的肽靶向的膜受体的选择性表达。使用这种转录靶向方法可以限制肿瘤的基因表达并限制体内正常组织中沉积的辐射剂量。
Radioimmunotherapy is limited by a variety of factors, including poor tumor penetration of monoclonal antibodies and low levels of intratumoral antigen expression. To address these limitations, a gene therapy strategy was devised to genetically induce tumor cells to express enhanced levels of membrane receptors with high affinity for a radiolabeled peptide. We designated this approach as genetic radioisotope targeting strategy. To this end, an adenoviral vector (AdCMVGRPr) encoding the murine gastrin-releasing peptide receptor (GRPr) was used to achieve a high level of binding of radiolabeled bombesin (BBN). To achieve genetic induction of membrane GRPr specifically to tumor cells, we constructed two adenoviral vectors encoding the GRPr gene under the control of the tumor-specific regulatory elements, DF3 (AdDF3GRPr) or erbB-2 (AderbGRPr). We investigated the binding of [I-125]BBN to the GRPr following infection with AdDF3GRPr and AderbGRPr in a panel of human breast, pancreatic, and cholangiocarcinoma tumor cell lines, [I-125]BBN binding and GRPr expression increased with increasing multiplicities of infection of AdCMVGRPr in all of the cell lines tested. Breast cancer cell lines expressing erbB-2 showed significant GRPr expression using AderbGRPr. A similar result was observed in breast and cholangiocarcinoma cells infected with AdDF3GRPr expressing MUC1 as detected by immunohistochemistry but was not seen in the pancreatic cell lines tested. Thus, adenoviral vectors with tissue-specific promoter elements can be used to achieve a selective expression of membrane receptors that can be targeted with a radiolabeled peptide. The use of such a transcriptional targeting approach may restrict gene expression to tumors and limit the radiation dose deposited in normal tissues in vivo.