Combined p14ARF and Interferon-beta Gene Transfer to the Human Melanoma Cell Line SK-MEL-147 Promotes Oncolysis and Immune Activation

Autor(es)

Cerqueira, Otto Luiz Dutra ^[1] ; Clavijo-Salomon, Maria Alejandra ^{[2, 1]} ; Cardoso, Elaine Cristina ^[3] ; Citrangulo Tortelli Junior, Tharcisio ^[1] ; Mendonca, Samir Andrade ^[1] ; Barbuto, Jose Alexandre M. ^{[2, 4]} ; Strauss, Bryan E. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo FMUSP, Fac Med, Ctr Invest Translac Oncol CTO, Inst Canc Estado Sao Paulo ICESP, Sao Paulo - Brazil

^[2] Univ Sao Paulo, Dept Imunol, Inst Ciencias Biomed, Sao Paulo - Brazil

^[3] Univ Sao Paulo FMUSP, Fac Med, Dept Pediat, Sao Paulo - Brazil

^[4] Univ Sao Paulo, Fac Med, Lab Med Invest Pathogenesis & Targeted Therapy On, Dept Hematol, Hosp Clin HCFMUSP, Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: FRONTIERS IN IMMUNOLOGY; v. 11, OCT 22 2020.
• Citações Web of Science: 0

Resumo

Immune evasion is an important cancer hallmark and the understanding of its mechanisms has generated successful therapeutic approaches. Induction of immunogenic cell death (ICD) is expected to attract immune cell populations that promote innate and adaptive immune responses. Here, we present a critical advance for our adenovirus-mediated gene therapy approach, where the combined p14ARF and human interferon-beta (IFN beta) gene transfer to human melanoma cells led to oncolysis, ICD and subsequent activation of immune cells. Our results indicate that IFN beta alone or in combination with p14ARF was able to induce massive cell death in the human melanoma cell line SK-MEL-147, though caspase 3/7 activation was not essential. In situ gene therapy of s.c. SK-MEL-147 tumors in Nod-Scid mice revealed inhibition of tumor growth and increased survival in response to IFN beta alone or in combination with p14ARF. Emission of critical markers of ICD (exposition of calreticulin, secretion of ATP and IFN beta) was stronger when cells were treated with combined p14ARF and IFN beta gene transfer. Co-culture of previously transduced SK-MEL-147 cells with monocyte-derived dendritic cells (Mo-DCs) derived from healthy donors resulted in increased levels of activation markers HLA-DR, CD80, and CD86. Activated Mo-DCs were able to prime autologous and allogeneic T cells, resulting in increased secretion of IFN gamma, TNF-alpha, and IL-10. Preliminary data showed that T cells primed by Mo-DCs activated with p14ARF+IFN beta-transduced SK-MEL-147 cells were able to induce the loss of viability of fresh non-transduced SK-MEL-147 cells, suggesting the induction of a specific cytotoxic population that recognized and killed SK-MEL-147 cells. Collectively, our results indicate that p14ARF and IFN beta delivered by our adenoviral system induced oncolysis in human melanoma cells accompanied by adaptive immune response activation and regulation. (AU)

• Processo FAPESP: 17/13686-9 - Planejamento racional de um agonista de STING como modelo para a implementação de uma plataforma de desenvolvimento de novas imunoterapias contra câncer
• Beneficiário: Maria Alejandra Clavijo Salomón
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 17/23068-0 - Avaliação da transferência gênica de p14Arf e IFN-beta em melanoma primário humano buscando evidências ex vivo de resposta imunológica e eficácia terapêutica in vivo
• Beneficiário: Otto Luiz Dutra Cerqueira
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 15/26580-9 - Terapia gênica do câncer: alinhamento estratégico para estudos translacionais
• Beneficiário: Bryan Eric Strauss
• Modalidade de apoio: Auxílio à Pesquisa - Temático