EGFR-targeted Chimeras of Pseudomonas ToxA released into the extracellular milieu by attenuated Salmonella selectively kill tumor cells.
EGFR-targeted Chimeras of Pseudomonas ToxA released into the extracellular milieu by attenuated Salmonella selectively kill tumor cells.
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DOI：
10.1002/bit.26026
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发表时间：
2016-12
影响因子：
3.8
通讯作者：
Bermudes, David
中科院分区：
文献类型：
--
作者：
Quintero, David;Carrafa, Jamie;Vincent, Lena;Bermudes, David
肿瘤靶向沙门氏菌VNP20009在小鼠肿瘤模型中优先在肿瘤组织内复制并部分抑制肿瘤生长。这些沙门氏菌在与癌细胞直接接触时具有局部诱导细胞凋亡的能力，但它们缺乏显著的旁观者杀伤作用，这可能与它们在人类临床研究中总体缺乏抗肿瘤活性有关。为了在不增强整体毒性的情况下弥补这一缺陷，我们通过工程使细菌表达表皮生长因子受体(EGFR)靶向的细胞毒蛋白，这些蛋白被释放到细胞外环境中。在这项研究中，我们证明了沙门氏菌株VNP20009能够产生三种不同形式的假单胞菌外毒素A(TOXA)与肿瘤生长因子α(α)的嵌合，从而导致其产生具有细胞毒性的培养上清，并通过四唑盐还原试验、罗丹明123和JC-10线粒体去极化试验检测其诱导EGFR阳性癌细胞凋亡的作用。此外，与野生型毒素相比，Toxa REDLK内质网滞留信号与KDEL的交换以及ColE3裂解蛋白的共表达导致了总体上更高的细胞毒性。这种方法有可能显著增强VNP20009的抗肿瘤活性，同时保持其先前建立的安全性。生物技术。比昂斯。2016；113：2698-2711。©2016作者。生物技术和生物工程由威利期刊出版公司出版。
Tumor‐targeted Salmonella VNP20009 preferentially replicate within tumor tissue and partially suppress tumor growth in murine tumor models. These Salmonella have the ability to locally induce apoptosis when they are in direct contact with cancer cells but they lack significant bystander killing, which may correlate with their overall lack of antitumor activity in human clinical studies. In order to compensate for this deficiency without enhancing overall toxicity, we engineered the bacteria to express epidermal growth factor receptor (EGFR)‐targeted cytotoxic proteins that are released into the extracellular milieu. In this study, we demonstrate the ability of the Salmonella strain VNP20009 to produce three different forms of the Pseudomonas exotoxin A (ToxA) chimeric with a tumor growth factor alpha (TGFα) which results in its producing culture supernatants that are cytotoxic and induce apoptosis in EGFR positive cancer cells as measured by the tetrazolium dye reduction, and Rhodamine 123 and JC‐10 mitochondrial depolarization assays. In addition, exchange of the ToxA REDLK endoplasmic reticulum retention signal for KDEL and co‐expression of the ColE3 lysis protein resulted in an overall increased cytotoxicity compared to the wild type toxin. This approach has the potential to significantly enhance the antitumor activity of VNP20009 while maintaining its previously established safety profile. Biotechnol. Bioeng. 2016;113: 2698–2711. © 2016 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals, Inc.
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