use of tissue-specific microrna to control pathology of wild-type adenovirus without attenuation of its ability to kill cancer cells使用组织微控制病理无衰减的野生型腺病毒杀死癌细胞的能力.pdfVIP

Use of Tissue-Specific MicroRNA to Control Pathology of Wild-Type Adenovirus without Attenuation of Its Ability to Kill Cancer Cells 1 1 1 1 2 Ryan Cawood , Hannah H. Chen , Fionnadh Carroll , Miriam Bazan-Peregrino , Nico van Rooijen , Leonard W. Seymour1* 1 Department of Clinical Pharmacology, University of Oxford, Oxford, United Kingdom, 2 Department of Molecular Cell Biology, Vrije Universiteit, Amsterdam, The Netherlands Abstract 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 39 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 61010