Glioblastoma (GBM) is the most common brain tumor in adults. The average survival at best standard care including surgery and radio-chemotherapy is approximately 20 months. In several pre- and clinical trials, it has been shown that oncolytic virotherapy is a safe approach to treat solid tumors, and it has been proven to be efficacious in different settings of GBM and other cancers. It is however limited to an intratumoral virus application due to the complexity of the immune system, leading to a fast inactivation of systemically applied viruses. In our study we use the oncolytic adenovirus XVir-N-31 in a xenograft, orthotopic mouse GBM model. As an alternative to intratumoral virus injection we examined the therapeutic effects of XVir-N-31 using a novel approach. We intranasally apply XVir-N-31 loaded, optimized “virus-shuttle-cells” to GBM-bearing mice. Intranasal delivery of drugs or therapeutic cells is a non-invasive, effective and less tedious approach to target brain disease and, in our approach, also brain tumors. In our study we examine the feasibility and therapeutic efficacy of this approach using XVir-N-31-loaded hepatic stellate cells. We hypothesize that this treatment strategy will also target infiltrative and invasively growing glioma cells that are distant from the original tumor and that cannot be targeted by a standard intratumoral OAV injection. Additionally, in a second step we will define the therapeutic window and efficacy of combining intranasal delivery of XVir-N-31 loaded shuttle cells with a single intratumoral injection of this oncolytic virus in glioma bearing mice.