Tutor: Dott.ssa Tatiana Gianni (DIMES) e Prof.ssa Anna Zaghini (DIMEVET)
TITOLO DEL PROGETTO
Oncolytic Herpes Viruses retargeted to cancer-specific receptors: set-up of an immunocompetent mouse model
ABSTRACT DEL PROGETTO
To enter cell, human herpes simplex virus (HSV) use both viral fusion glycoproteins gH/gL/gB and viral glycoprotein gD that is responsible for cell/tissue tropism. Glycoprotein gD has been previously engineered with a single chain antibody specific for the HER2 receptor that is overexpressed by some types of cancer cells. The recombinant virus enters cells through the HER2 receptor only and has lost the ability to infect the cells that are the natural targets of HSV infection. This oncolytic, HER-2 retargeted virus (o-HSV-HER2ret) has been shown to be able to efficiently kill cancer cells both in vitro and in vivo models. However pre-clinical studies and clinical trials testing oncolytic viruses have demonstrated that, alone, the lytic activity of the virus against cancer cells is not sufficient for tumor clearance. An induction of an immune response is pivotal to give rise to a complete, immune-mediated tumor regression. The tumor micro-environment is immunosuppressive and cancer cells have devised strategies to evade the action of the immune system. As such,we have insertedimmune stimulatory molecules intoo-HSV-HER2ret.The rationale of using oncolytic viruses is to try to stimulate an initial immune response against tumor antigens. If the oncolytic virus is also engineered to express immune stimulatory molecules, the viral infection inside the tumor plus the aid of immune stimulation could be potent enough to overcome immune tolerance, leading to the development of an effective and long lasting immune response against cancer cells. In order to test these immune stimulatory o-HSVs, an immunocompetent mouse model, the C57Bl/6 huHER2 mouse strain, tolerant for the human HER2 protein, will be employed.
PIANO DELLE ATTIVITÀ DELL’ASSEGNISTA
The research fellow will carry out all the laboratory work required to maintain and expand the C57Bl/6 huHER2 colony. More specifically, he/she will help in the general husbandry to maintain the colony, will plan and follow the breeding, will wean and mark the pups, will genotype the mice. Both a subcutaneous tumor model and a systemic tumor model will be established using different doses of congenic huHER2+ cancer cell lines. Mice will be monitored after the tumor induction and the tumor mass will be measured. Administration of the oncolytic virus will be performed both locally and systemically. Harvesting and preparation of organs and serum will be carried out at the end of the experiment to proceed with analysis of the tumor and the tumor-specific immune response.
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