Use of tissue-specific microRNA to control pathology of wild-type adenovirus without attenuation of its ability to kill cancer cells.
Use of tissue-specific microRNA to control pathology of wild-type adenovirus without attenuation of its ability to kill cancer cells.
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DOI：
10.1371/journal.ppat.1000440
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发表时间：
2009-05
期刊：
影响因子：
6.7
通讯作者：
Seymour LW
中科院分区：
文献类型：
--
作者：
Cawood R;Chen HH;Carroll F;Bazan-Peregrino M;van Rooijen N;Seymour LW
复制病毒在生物医学中有着广泛的应用，特别是在癌症病毒治疗和减毒疫苗的设计中;然而，在脆弱组织中不受控制的病毒复制可能导致病理学，并且通常限制了强效毒株的使用。组织选择性microRNA表达的知识增加，现在提供了工程复制病毒的可能性，这些病毒在潜在病理学位点的RNA水平上减毒，但在不表达相关microRNA的位点保留野生型复制活性。为了评估这种方法对DNA病毒腺病毒的有用性，我们设计了一种肝细胞安全的野生型腺病毒5（Ad 5），它通常介导显着的毒性，并在小鼠中可能是致命的。为此，我们在E1 A转录盒的3′ UTR中包含了肝细胞选择性microRNA mir-122的结合位点。通过将E1 A与荧光素酶融合产生的这些病毒的成像版本显示，在静脉内递送给小鼠后，包含mir-122结合位点导致E1 A的肝脏表达降低高达80倍。给予10倍致死剂量的野生型Ad 5（5×1010病毒颗粒/小鼠）的动物显示出大量的肝脏基因组复制和广泛的肝脏病理学，而包含4个microRNA结合位点使复制减少50倍，几乎消除了肝脏毒性。这种修饰的野生型病毒在癌细胞内保留了全部活性，并提供了一种有效的、对肝脏安全的溶瘤病毒。除了为癌症病毒治疗提供许多有效的新病毒外，病毒复制的microRNA控制还应该为设计应用于广泛病毒疾病的安全减毒疫苗提供新策略。减毒病毒在医学上有重要的应用，包括用作疫苗（特别是麻疹、腮腺炎、脊髓灰质炎、流感和水痘），以及作为选择性抗癌剂的实验性开发，即所谓的“病毒疗法”。在这两种情况下，野生型病毒通常最有效;然而，减毒病毒通常被开发用于降低重大病毒病理学的风险。最近在理解microRNA对基因表达的调节方面的进展现在提供了设计在潜在病理学位点“选择性减毒”的病毒的可能性，通过工程化它们以被在那里表达的microRNA分子抑制。在这里，我们设计了野生型腺病毒，用于识别肝细胞中表达的microRNA，产生一种病毒，该病毒保留野生型感染和复制的治疗活性位点（如癌细胞），但在肝细胞中严重减毒，无论是在体外还是体内。即使以野生型病毒致死剂量的十倍施用，这种病毒也不会对小鼠造成显著的肝毒性。生产具有复制能力的病毒并去除关键毒性的能力应该为开发改进的癌症治疗和更好的疫苗提供新的平台。
Replicating viruses have broad applications in biomedicine, notably in cancer virotherapy and in the design of attenuated vaccines; however, uncontrolled virus replication in vulnerable tissues can give pathology and often restricts the use of potent strains. Increased knowledge of tissue-selective microRNA expression now affords the possibility of engineering replicating viruses that are attenuated at the RNA level in sites of potential pathology, but retain wild-type replication activity at sites not expressing the relevant microRNA. To assess the usefulness of this approach for the DNA virus adenovirus, we have engineered a hepatocyte-safe wild-type adenovirus 5 (Ad5), which normally mediates significant toxicity and is potentially lethal in mice. To do this, we have included binding sites for hepatocyte-selective microRNA mir-122 within the 3′ UTR of the E1A transcription cassette. Imaging versions of these viruses, produced by fusing E1A with luciferase, showed that inclusion of mir-122 binding sites caused up to 80-fold decreased hepatic expression of E1A following intravenous delivery to mice. Animals administered a ten-times lethal dose of wild-type Ad5 (5×1010 viral particles/mouse) showed substantial hepatic genome replication and extensive liver pathology, while inclusion of 4 microRNA binding sites decreased replication 50-fold and virtually abrogated liver toxicity. This modified wild-type virus retained full activity within cancer cells and provided a potent, liver-safe oncolytic virus. In addition to providing many potent new viruses for cancer virotherapy, microRNA control of virus replication should provide a new strategy for designing safe attenuated vaccines applied across a broad range of viral diseases. Attenuated viruses have found important applications in medicine, including their use as vaccines (notably for measles, mumps, polio, influenza, and chicken pox) and their experimental development as selective cancer-killing agents, so-called “virotherapy.” Wild-type versions are often most effective in both of these settings; however, attenuated viruses have usually been developed to decrease the risk of significant viral pathology. Recent advances in understanding regulation of gene expression by microRNA now afford the possibility to design viruses that are “selectively attenuated” in sites of potential pathology, by engineering them for inhibition by microRNA molecules that are expressed there. Here we have engineered wild-type adenovirus for recognition by a microRNA expressed in hepatocytes, producing a virus that retains wild-type infection and replication at sites of therapeutic activity (such as cancer cells) but is severely attenuated in hepatocytes, both in vitro and in vivo. This virus caused no significant liver toxicity to mice even when applied at ten times the lethal dose of wild-type virus. The ability to produce replication-competent viruses with key toxicities removed should provide a new platform for development of improved cancer treatments and better vaccines for a broad range of viral diseases.
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影响因子：
56.9
作者：
Lagos-Quintana, M;Rauhut, R;Tuschl, T
通讯作者：
Tuschl, T
影响因子：
30.3
作者：
Barnes, Dwight;Kunitomi, Mark;Vignuzzi, Marco;Saksela, Kalle;Andino, Raul
通讯作者：
Andino, Raul
影响因子：
82.9
作者：
Brown, Brian D.;Venneri, Mary Anna;Naldini, Luigi
通讯作者：
Naldini, Luigi
影响因子：
4.1
作者：
Chang, Jinhong;Nicolas, Emmanuelle;Taylor, John M
通讯作者：
Taylor, John M
DOI：
10.1007/bf01863914
发表时间：
1931-01-01
影响因子：
--
作者：
Karber, G
通讯作者：
Karber, G