Detection of neuroendocrine tumors using promoter-specific secreted Gaussia luciferase
Detection of neuroendocrine tumors using promoter-specific secreted Gaussia luciferase
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DOI：
10.3892/ijo.2015.3223
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发表时间：
2016-01-01
影响因子：
5.2
通讯作者：
Lan, Michael S.
中科院分区：
文献类型：
--
作者：
Tseng, Alan Wei-Shun;Akerstrom, Victoria;Lan, Michael S.
神经内分泌（NE）肿瘤的准确检测对于患者更好的预后和治疗效果至关重要。为了证明使用腺病毒载体检测NE肿瘤的有效性，我们构建了一对腺病毒载体，它们结合在一起可以在感染NE肿瘤后有条件地复制并将高斯荧光素酶释放到循环中。这两个载体的表达由insm1启动子（胰岛素瘤相关-1）调控，该启动子在NE肿瘤和发育中的NE组织中特别活跃，但在正常成人组织中沉默。为了保留INSM1启动子的肿瘤特异性，我们利用鸡β -珠蛋白HS4绝缘子的核心绝缘子序列和神经元限制性沉默元件（NRSE）对该启动子进行了修饰。这种修饰的insm1启动子可以在腺病毒构建体中保持NE肿瘤特异性，同时驱动突变的腺病毒E1A基因（Delta 24E1A）、Metridia或Gaussia荧光素酶基因。体外细胞系和小鼠异种移植人肿瘤研究显示，insm1启动子在NE肺癌、神经母细胞瘤、髓母细胞瘤、视网膜母细胞瘤和胰岛素瘤中具有NE特异性。当我们将insm1启动子驱动的高斯荧光素酶与Delta 24E1A结合使用时，与单独使用insm1 -p -高斯病毒相比，共感染的NE肿瘤分泌更高水平的高斯荧光素酶。在小鼠皮下异种移植瘤模型中，联合病毒在感染insm1阳性的NE肺肿瘤>= 12天后分泌出可检测水平的高斯荧光素酶。因此，insm1启动子特异性条件复制腺病毒是一种敏感的诊断工具，可以帮助临床医生检测NE肿瘤。
Accurate detection of neuroendocrine (NE) tumors is critically important for better prognosis and treatment outcomes in patients. To demonstrate the efficacy of using an adenoviral vector for the detection of NE tumors, we have constructed a pair of adenoviral vectors which, in combination, can conditionally replicate and release Gaussia luciferase into the circulation after infecting the NE tumors. The expression of these two vectors is regulated upstream by an INSM1-promoter (insulinoma-associated-1) that is specifically active in NE tumors and developing NE tissues, but silenced in normal adult tissues. In order to retain the tumor-specificity of the INSM1 promoter, we have modified the promoter using the core insulator sequence from the chicken beta-globin HS4 insulator and the neuronal restrictive silencing element (NRSE). This modified INSM1-promoter can retain NE tumor specificity in an adenoviral construct while driving a mutated adenovirus E1A gene (Delta 24E1A), the Metridia, or Gaussia luciferase gene. The in vitro cell line and mouse xenograft human tumor studies revealed the NE specificity of the INSM1-promoter in NE lung cancer, neuroblastoma, medulloblastoma, retinoblastoma, and insulinoma. When we combined the INSM1-promoter driven Gaussia luciferase with Delta 24E1A, the co-infected NE tumor secreted higher levels of Gaussia luciferase as compared to the INSM1p-Gaussia virus alone. In a mouse subcutaneous xenograft tumor model, the combination viruses secreted detectable level of Gaussia luciferase after infecting an INSM1-positive NE lung tumor for >= 12 days. Therefore, the INSM1-promoter specific conditional replicating adenovirus represents a sensitive diagnostic tool to aid clinicians in the detection of NE tumors.