Engineered adenoviruses combine enhanced oncolysis with improved virus production by mesenchymal stromal carrier cells
Engineered adenoviruses combine enhanced oncolysis with improved virus production by mesenchymal stromal carrier cells
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DOI：
10.1002/ijc.29442
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发表时间：
2015-08-15
影响因子：
6.4
通讯作者：
Nettelbeck, Dirk M.
中科院分区：
文献类型：
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作者：
Hammer, Katharina;Kazcorowski, Adam;Nettelbeck, Dirk M.
溶瘤病毒已在临床前和临床研究中证明了其安全性和独特的多效性肿瘤破坏活性。然而，它们的输送会受到血液成分灭活病毒和隔离到健康组织的影响。因此，间充质基质细胞（MSC）已被用作载体细胞，用于在溶瘤病毒离体感染后将屏蔽病毒递送至肿瘤。然而，间充质干细胞的感染和颗粒产生仍不令人满意。在这里，我们报告了工程化的溶瘤腺病毒（OAd），用于改善 MSC 的病毒生产和递送。 OAd 独特地适合分子工程，这有助于改善肿瘤细胞的破坏。但对于 MSC 介导的治疗方案，OAd 工程需要在 MSC 和肿瘤细胞中实现有效的感染和复制。我们发现，含有腺病毒血清型 3 细胞结合结构域的 Ad5/3 嵌合 OAd 衣壳可强烈增加进入人骨髓源性 MSC 以及已建立的原发性胰腺癌细胞的能力。此外，我们还发现，通过删除抗凋亡病毒基因 E1B19K 或表达死亡配体 TRAIL，具有工程化后进入功能的 OAd 显着增加了 MSC 释放的病毒滴度，而 MSC 迁移并未受到阻碍。最后，这些病毒修饰或用于局部 5-FC 前药激活的 FCU1 病毒表达，改善了肿瘤细胞杀伤，在一组胰腺癌细胞培养物中实现了互补的细胞毒性特征。总之，我们的研究建立了 OAd 复制的进入后修饰，以改善载体细胞的病毒传递，并为胰腺癌个性化治疗的未来临床开发提出了一组优化的 OAd。
Oncolytic viruses have demonstrated in pre-clinical and clinical studies safety and a unique pleiotropic activity profile of tumor destruction. Yet, their delivery suffers from virus inactivation by blood components and sequestration to healthy tissues. Therefore, mesenchymal stromal cells (MSCs) have been applied as carrier cells for shielded virus delivery to tumors after ex vivo infection with oncolytic viruses. However, infection and particle production by MSCs have remained unsatisfying. Here, we report engineered oncolytic adenoviruses (OAds) for improved virus production and delivery by MSCs. OAds are uniquely amenable to molecular engineering, which has facilitated improved tumor cell destruction. But for MSC-mediated regimens, OAd engineering needs to achieve efficient infection and replication in both MSCs and tumor cells. We show that an Ad5/3 chimeric OAd capsid, containing the adenovirus serotype 3 cell-binding domain, strongly increases the entry into human bone marrow-derived MSCs and into established and primary pancreatic cancer cells. Further, we reveal that OAd with engineered post-entry functions-by deletion of the anti-apoptotic viral gene E1B19K or expression of the death ligand TRAIL-markedly increased virus titers released from MSCs, while MSC migration was not hampered. Finally, these virus modifications, or viral expression of FCU1 for local 5-FC prodrug activation, improved tumor cell killing implementing complementary cytotoxicity profiles in a panel of pancreatic cancer cell cultures. Together, our study establishes post-entry modification of OAd replication for improving virus delivery by carrier cells and suggests a panel of optimized OAds for future clinical development in personalized treatment of pancreatic cancer.