While cancer immunotherapy has gained much deserved attention in recent years, many areas regarding the optimization of such modalities remain unexplored, including the development of novel approaches and the strategic combination of therapies that target multiple aspects of the cancer-immunity cycle. Our own work involves the use of gene transfer technology to promote cell death and immune stimulation. Such immunogenic cell death, mediated by the combined transfer of the alternate reading frame (p14ARF in humans and p19Arf in mice) and the interferon-β cDNA in our case, was shown to promote an antitumor immune response in mouse models of melanoma and lung carcinoma. With these encouraging results, we are now setting out on the road toward translational and preclinical development of our novel immunotherapeutic approach. Here, we outline the perspectives and challenges that we face, including the use of human tumor and immune cells to verify the response seen in mouse models and the incorporation of clinically relevant models, such as patient-derived xenografts and spontaneous tumors in animals. In addition, we seek to combine our immunotherapeutic approach with other treatments, such as chemotherapy or checkpoint blockade, with the goal of reducing dosage and increasing efficacy. The success of any translational research requires the cooperation of a multidisciplinary team of professionals involved in laboratory and clinical research, a relationship that is fostered at the Cancer Institute of Sao Paulo.

pelos ensinamentos, discussões, ideias, direcionamentos e por estar SEMPRE presente. Por ter me recebido em seu grupo, concedido essa incrível oportunidade e por ter confiado em mim. Obrigada por ter me tornado a pesquisadora que sou hoje.Aos alunos e funcionários do ICESP e da Faculdade de Medicina da USP pela incessante colaboração. Á FAPESP pelo financiamento imprescindível para a elaboração e desenvolvimento deste projeto.Aos professores Roger Chammas e Luisa Villa, por serem exemplos de profissionais que eu desejo alcançar.Aos pesquisadores e amigos Daniela, Rodrigo, Marlous, Paulo, Samir, Aline, Igor, Otto, Ruan, Nayara, Nadine, Fernanda e Otavio pelo treinamento, apoio, conselhos, discussões e cafés. Sem vocês este projeto não seria possível. pela colaboração e excelentes discussões e ideias. À minha amiga e sócia Simone Crestoni, pelo incentivo, apoio, por suportar minhas ausências tantas vezes, por ter sido a responsável por abrir as portas para essa jornada e por continuar me fazendo amar a pesquisa. À minha família Marlene, Osmar, Alessandra e Rodrigo por todo amor, carinho, apoio incondicionais, sempre. Ao meu marido Eduardo, pelo amor, compreensão, companheirismo e pela oportunidade maravilhosa de amá-lo. gene TP53 e a avaliação da expressão de genes da via de TP53 induzidos por doxorrubicina e Nutlin-3. RESULTADOS: As quatro linhagens de melanoma canino estabelecidas possuem capacidade tumorigênica e de serem transduzidas pelos vetores adenovirais. Não foram identificadas alterações na sequência do TP53 na região avaliada e a doxorrubicina promoveu aumento da expressão dos transgenes dirigidos pelo promotor PG e a ativação de genes da via de TP53. CONCLUSÕES: A funcionalidade desta plataforma adenoviral aprimorada abre oportunidades para estudos da transferência gênica, incluindo da combinação p19ARF/IFNβ, nas linhagens estabelecidas. Com o sucesso destas análises, teremos um importante modelo experimental a ser utilizado no desenvolvimento de novas terapias para o melanoma. ABSTRACT Silva GRO. Establishment of canine melanoma cell lines and transduction with improved adenoviral vectors [dissertation]. São Paulo: "Faculdade de Medicina, Universidade de São Paulo"; 2019. INTRODUCTION: Melanoma is a cancer of high mortality both in human and veterinary medicine due to the poor response to therapies and the metastatic evolution of the disease. Advances in the field of oncology have shown that agents that target immune system components, as well as molecularly targeted therapies, are very promising for the treatment of melanoma. Our group has developed viral vectors for gene transfer of antitumor factors. The improvement in the adenoviral vector includes the insertion of the RGD tripeptide, which allows a broad transduction tropism, and the use of a p53 responsive promoter to control therapeutic gene expression. Canine melanoma may be considered as an experimental model since it is a spontaneous cancer with biological behavior similar to human melanoma, thus we aim to test our approach, which we call Ad...

Immunotherapy of cancer has deservedly gained much attention in the past few years and is likely to continue to advance and become a fundamental cancer treatment. While vaccines, chimeric antigen receptor (CAR) T cells and checkpoint blockade have received the lion's share of the attention, an important direct role for gene transfer as an immunotherapy is emerging. For example, oncolytic viruses induce immunogenic cell death, thus liberating both antigens and the signals that are necessary for the activation of antigenpresenting cells, ensuring stimulation of an adaptive response. In another example, transfer of prodrug converting enzymes, such as the herpes simplex virus-thymidine kinase (HSV-tk) gene or the cytosine deaminase gene, has been shown to promote an immune response, thus functioning as immunotherapies. Alternatively, our own work involves the use of nonreplicating viral vectors for the simultaneous delivery of gene combinations that promote both cell death and an immune response. In fact, our gene transfer approach has been applied as a vaccine, immunotherapy or in situ gene therapy, resulting in immunogenic cell death and the induction of a protective immune response. Here, we highlight the development of these approaches both in terms of technical advances and clinical experience.