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Oncolytic adenovirus expressing therapeutic transgenes represent a promising alternative treatment for GBM. Of the many human adenoviral serotypes described to date, adenovirus 5 (HAdV-C5) has been the most utilised clinically and experimentally. However, the use of Ad5 as an anti-cancer agent may be hampered by naturally high seroprevalence rates to HAdV-C5 coupled with the infection of healthy cells via native receptors. To explore whether alternative natural adenoviral tropisms are better suited to GBM therapeutics, we pseudotyped an HAdV-C5-based platform using the fibre knob protein from alternative serotypes. We demonstrate that the adenoviral entry receptor coxsackie, adenovirus receptor (CAR) and CD46 are highly expressed by both GBM and healthy brain tissue, whereas Desmoglein 2 (DSG2) is expressed at a low level in GBM. We demonstrate that adenoviral pseudotypes, engaging CAR, CD46 and DSG2, effectively transduce GBM cells. However, the presence of these receptors on non-transformed cells presents the possibility of off-target effects and therapeutic transgene expression in healthy cells. To enhance the specificity of transgene expression to GBM, we assessed the potential for tumour-specific promoters hTERT and survivin to drive reporter gene expression selectively in GBM cell lines. We demonstrate tight GBM-specific transgene expression using these constructs, indicating that the combination of pseudotyping and tumour-specific promoter approaches may enable the development of efficacious therapies better suited to GBM.<\/jats:p>","DOI":"10.3390\/v15051086","type":"journal-article","created":{"date-parts":[[2023,5,1]],"date-time":"2023-05-01T12:02:31Z","timestamp":1682942551000},"page":"1086","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Engineering Adenoviral Vectors with Improved GBM Selectivity"],"prefix":"10.3390","volume":"15","author":[{"given":"Emily A.","family":"Bates","sequence":"first","affiliation":[{"name":"Division of Cancer and Genetics, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2606-8930","authenticated-orcid":false,"given":"Charlotte","family":"Lovatt","sequence":"additional","affiliation":[{"name":"Division of Cancer and Genetics, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alice R.","family":"Plein","sequence":"additional","affiliation":[{"name":"Division of Cancer and Genetics, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"James A.","family":"Davies","sequence":"additional","affiliation":[{"name":"Division of Cancer and Genetics, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8411-8775","authenticated-orcid":false,"given":"Florian A.","family":"Siebzehnrubl","sequence":"additional","affiliation":[{"name":"European Cancer Stem Cell Research Institute, School of Biosciences, Cardiff University, Maindy Road, Cardiff CF24 4HQ, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9302-1761","authenticated-orcid":false,"given":"Alan L.","family":"Parker","sequence":"additional","affiliation":[{"name":"Division of Cancer and Genetics, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, UK"},{"name":"Systems Immunity University Research Institute, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,4,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"iv1","DOI":"10.1093\/neuonc\/nou223","article-title":"CBTRUS statistical report: Primary brain and central nervous system tumors diagnosed in the United States in 2007\u20132011","volume":"16","author":"Ostrom","year":"2014","journal-title":"Neuro-Oncology"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1007\/s10143-017-0928-7","article-title":"Multiple high-grade gliomas: Epidemiology, management, and outcome: A systematic review and meta-analysis","volume":"42","author":"Cagnazzo","year":"2019","journal-title":"Neurosurg. 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