Vaccines have become an important approach for the treatment of cancer. Cancer vaccines help the immune system to detect and eradicate tumor cells. Also, cancer vaccines are designed to stimulate an effective immune response that can create long-term immune memory to prevent tumor recurrence. This treatment approach involves the administration of a vaccine comprising or encoding an antigen and can often be combined with an adjuvant to further promote the immune response. The goal of this research was to study the effect of the polyanhydride composition of prophylactic cancer vaccine formulations on the tumor-specific immune response. To achieve this goal, three different amphiphilic polyanhydride copolymers were generated comprising different ratios of 1,6-bis-(p-carboxyphenoxy)-hexane (CPH) and 1,8-bis-(p-carboxyphenoxy)-3,6-dioxaoctane (CPTEG) or sebacic anhydride (SA) monomers. These copolymers were used to fabricate particles encapsulating a model antigen, ovalbumin (OVA), using a double emulsion solvent evaporation technique. The ability of the three different compositions of amphiphilic polyanhydride copolymers (50:50 CPTEG:CPH, 20:80 CPTEG:CPH, and 20:80 CPH:SA) encapsulating OVA to elicit immune responses was investigated. Further, the impact of soluble unmethylated oligodeoxynucleotides containing deoxycytidyl-deoxyguanosine dinucleotides (CpG ODN), an immunologic adjuvant, on the immune response to the three formulations was also studied. The immune response to cancer vaccines was measured after treatment of C57BL/6J mice with two subcutaneous injections, seven days apart, of 50 μg OVA encapsulated in particles composed of different polyanhydride copolymers with or without 25 μg CpG ODN. In vivo studies showed that 20:80 CPTEG:CPH particles encapsulating OVA significantly stimulated the highest level of CD8+ T lymphocytes, generated the highest serum titers of OVA-specific IgG antibodies, and produced longer survival in comparison to formulations involving the other polyanhydride copolymers. The results also revealed that supplementing the vaccine formulations with CpG ODN did not enhance the immunogenicity of OVA. These results accentuate the crucial role of the copolymer composition of polyanhydrides in stimulating the immune response and improving cancer vaccine efficacy.

The multifunctional signaling hub p62 is well recognized as a ubiquitin sensor and a selective autophagy receptor. As a ubiquitin sensor, p62 promotes NFκB activation by facilitating TRAF6 ubiquitination and aggregation. As a selective autophagy receptor, p62 sorts ubiquitinated substrates including p62 itself for lysosome-mediated degradation. p62 plays crucial roles in myriad cellular processes including DNA damage response, aging/senescence, infection and immunity, chronic inflammation, and cancerogenesis, dependent on or independent of autophagy. Targeting p62-mediated autophagy may represent a promising strategy for clinical interventions of different cancers. In this review, we summarize the transcriptional and post-translational regulation of p62, and its mechanistic roles in cancers, with the emphasis on its roles in regulation of DNA damage response and its connection to the cGAS-STING-mediated antitumor immune response, which is promising for cancer vaccine design.

Oncolytic viruses (OVs) are promising tumor-selective treatments, and the efficacy of OV therapies has been shown to depend heavily on the successful delivery and spread of these agents within the tumor mass to generate profound immunostimulatory effects. We have previously reported the potential of vanadium-based compounds such as vanadyl sulfate (VS) as immune-stimulatory enhancers of OV immunotherapy. These compounds, in conjunction with RNA-based OVs such as oncolytic VSVΔ51, improve viral spread and oncolysis, leading to long-term antitumor immunity and prolonged survival in resistant tumor models as previously reported. This effect is associated with a virus-induced antiviral type I IFN response shifting towards a type II IFN response. Here, the systemic impact and the relevant immunological changes following VS/VSVΔ51 combination therapy were investigated to understand the immunological mechanism of action leading to improved antitumor responses. We screened for the secretion of chemokines and cytokines in vivo to understand the mechanism of action regulating the recruitment of immune cells to the tumor in the CT26WT tumor model following treatment. Additionally, the antigen-specific immune response was investigated to further identify the relevant immunological changes following treatment with the VS+VSVΔ51 combination. Our data revealed that VS+VSVΔ51 combination therapy significantly increased the levels of IFN-γ and IL-6, and other key important pro-inflammatory cytokines and chemokines. Improved tumor antigen-specific T-cell responses were observed following the combined therapy. Supported by relevant immunological changes and as a proof of concept for the design of more effective therapeutic regimens, we found that local delivery of VSVΔ51 encoded with IL-12 or with other transgenes in combination with VS further improved therapeutic outcomes in a syngeneic CT26WT colon cancer model. We found that CD8+ T cells and Natural Killer (NK) cells play significant roles in establishing the therapeutic efficacy that we observed; Furthermore, engineering new and targeted therapeutic platforms to impact the antitumor immune response further improves the therapeutic benefits of the combined therapy.

A atividade física aeróbia reduz a incidência de diversos tipos de câncer e atenua o crescimento tumoral. Entretanto, existe uma lacuna na literatura sobre os mecanismos envolvidos nessa resposta. Dados recentes demonstram a importância da manutenção da função da resposta imune antitumoral para a atenuação da progressão da doença e estratégias capazes de aumentar essa resposta permanecem como um grande desafio. Diante disso, delineamos um estudo experimental para investigar especificamente a possível contribuição da atividade física aeróbia para a resposta imune antitumoral. Na presente dissertação, nós demonstramos que a atividade física aeróbia possui o potencial alterar a resposta de marcadores relacionados a resposta imune antitumoral. Em um primeiro estudo, observamos que a atividade física aeróbia atenuou o crescimento tumoral em três diferentes modelos de câncer em animais, assim como aumentou a sobrevida de animais com melanoma B16F10. Além disso, nós observamos que a atividade física aeróbia também altera a expressão gênica de marcadores de os linfócitos T infiltrantes de tumor (do inglês tumor infiltrating lymphocytes T - TILs-T) e de macrófagos associados ao tumor (do inglês tumor-associated macrophages - TAMs). Em segundo estudo, nosso objetivo foi explorar através de análises in silico respostas imunes tumorais que poderiam estar sendo moduladas pela atividade física aeróbia. Por meio da análise de Gene Set Enrichment Analysis (GSEA), observamos que a atividade física aeróbia prévia gerou uma assinatura imunológica tumoral semelhante à de pacientes com diferentes tipos de câncer e maior sobrevida. Essa assinatura imunológica revelou que pacientes com câncer de mama e melanoma que apresentam alta expressão gênica de BTG2, CD69, CFH, DUSP1 e PTGER4 em seus tumores, apresentam maior sobrevida em comparação aos pacientes com baixa expressão desses genes em seus tumores. A análise de GSEA também demonstrou que a atividade física aeróbia foi capaz de induzir uma assinatura imunológica semelhante à de animais com melanoma B16F10 sensíveis ao tratamento com o imunoterápico anti-CTLA-4 (do inglês anti- cytotoxic T-lymphocyte-associated antigen 4) e pacientes pós-tratamento com anti-CTLA-4. Essa assinatura imunológica também revelou que pacientes com melanoma que apresentam alta expressão gênica de PHC3, TET2, MACF1 e PARP8 em seus tumores, apresentam maior sobrevida em comparação aos pacientes com baixa expressão desses genes em seus tumores. Em conclusão, a atividade física aeróbia demonstra-se como uma potente ferramenta no combate a progressão da doença / It is already very well accepted that the aerobic physical activity reduces the incidence of many different types of cancer and mitigates the tumor progression. However, there is an investigation gap on the literature about the mechanisms underlying this response. Recently, a body of literature has arisen which show the importance of maintain antitumoral immune response to mitigated tumor progression, and strategy for enhance this response remains a major challenge. Presently, we demonstrate that the aerobic physical activity has the potential to modulate antitumor immune responses. Firstly, we have observed that the aerobic physical activity mitigated the tumor progression in three different animal models of cancer and increased survival in the animals which had B16F10 melanoma. Moreover, we have observed that the aerobic physical activity also seems modulate the tumor infiltrating T lymphocytes (TILs-T) and the tumor associated macrophages (TAMs) in as specific-tumor way. Based on the computer gene set enrichment analysis (GSEA), we have observed that the previous aerobic physical activity helped to develop an immunological signature of the tumor shared among patients with different cancer etiologies with higher survival over time. This signature introduced us to some genes that are associated with a higher survival over time when increased in tumors of patients with melanoma, breast cancer and lymphoma. The GSEA analysis also shows that the aerobic physical activity is able to introduce an immune signature similar to the one observed in animals with B16F10 melanoma sensitive to the immunotherapic anti- cytotoxic T-lymphocyte-associated antigen 4 (anti-CTLA-4) and patients post treatment with anti-CTLA-4. This immune signature also revealed genes with a strong association with high survival over time. This immune signature also revealed genes associated with a higher survival over time when their expression were increased in melanoma patients. In conclusion, aerobic physical activity is a powerful tool to counteract tumor progression due to its capacity to modulate antitumor immune responses

A atividade física aeróbia reduz a incidência de diversos tipos de câncer e atenua o crescimento tumoral. Entretanto, existe uma lacuna na literatura sobre os mecanismos envolvidos nessa resposta. Dados recentes demonstram a importância da manutenção da função da resposta imune antitumoral para a atenuação da progressão da doença e estratégias capazes de aumentar essa resposta permanecem como um grande desafio. Diante disso, delineamos um estudo experimental para investigar especificamente a possível contribuição da atividade física aeróbia para a resposta imune antitumoral. Na presente dissertação, nós demonstramos que a atividade física aeróbia possui o potencial alterar a resposta de marcadores relacionados a resposta imune antitumoral. Em um primeiro estudo, observamos que a atividade física aeróbia atenuou o crescimento tumoral em três diferentes modelos de câncer em animais, assim como aumentou a sobrevida de animais com melanoma B16F10. Além disso, nós observamos que a atividade física aeróbia também altera a expressão gênica de marcadores de os linfócitos T infiltrantes de tumor (do inglês tumor infiltrating lymphocytes T - TILs-T) e de macrófagos associados ao tumor (do inglês tumor-associated macrophages - TAMs). Em segundo estudo, nosso objetivo foi explorar através de análises in silico respostas imunes tumorais que poderiam estar sendo moduladas pela atividade física aeróbia. Por meio da análise de Gene Set Enrichment Analysis (GSEA), observamos que a atividade física aeróbia prévia gerou uma assinatura imunológica tumoral semelhante à de pacientes com diferentes tipos de câncer e maior sobrevida. Essa assinatura imunológica revelou que pacientes com câncer de mama e melanoma que apresentam alta expressão gênica de BTG2, CD69, CFH, DUSP1 e PTGER4 em seus tumores, apresentam maior sobrevida em comparação aos pacientes com baixa expressão desses genes em seus tumores. A análise de GSEA também demonstrou que a atividade física aeróbia foi capaz de induzir uma assinatura imunológica semelhante à de animais com melanoma B16F10 sensíveis ao tratamento com o imunoterápico anti-CTLA-4 (do inglês anti- cytotoxic T-lymphocyte-associated antigen 4) e pacientes pós-tratamento com anti-CTLA-4. Essa assinatura imunológica também revelou que pacientes com melanoma que apresentam alta expressão gênica de PHC3, TET2, MACF1 e PARP8 em seus tumores, apresentam maior sobrevida em comparação aos pacientes com baixa expressão desses genes em seus tumores. Em conclusão, a atividade física aeróbia demonstra-se como uma potente ferramenta no combate a progressão da doença / It is already very well accepted that the aerobic physical activity reduces the incidence of many different types of cancer and mitigates the tumor progression. However, there is an investigation gap on the literature about the mechanisms underlying this response. Recently, a body of literature has arisen which show the importance of maintain antitumoral immune response to mitigated tumor progression, and strategy for enhance this response remains a major challenge. Presently, we demonstrate that the aerobic physical activity has the potential to modulate antitumor immune responses. Firstly, we have observed that the aerobic physical activity mitigated the tumor progression in three different animal models of cancer and increased survival in the animals which had B16F10 melanoma. Moreover, we have observed that the aerobic physical activity also seems modulate the tumor infiltrating T lymphocytes (TILs-T) and the tumor associated macrophages (TAMs) in as specific-tumor way. Based on the computer gene set enrichment analysis (GSEA), we have observed that the previous aerobic physical activity helped to develop an immunological signature of the tumor shared among patients with different cancer etiologies with higher survival over time. This signature introduced us to some genes that are associated with a higher survival over time when increased in tumors of patients with melanoma, breast cancer and lymphoma. The GSEA analysis also shows that the aerobic physical activity is able to introduce an immune signature similar to the one observed in animals with B16F10 melanoma sensitive to the immunotherapic anti- cytotoxic T-lymphocyte-associated antigen 4 (anti-CTLA-4) and patients post treatment with anti-CTLA-4. This immune signature also revealed genes with a strong association with high survival over time. This immune signature also revealed genes associated with a higher survival over time when their expression were increased in melanoma patients. In conclusion, aerobic physical activity is a powerful tool to counteract tumor progression due to its capacity to modulate antitumor immune responses

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Baixas doses de agentes desmetilantes do DNA (DNMTi) tem efeitos antitumorais por induzirem parada do ciclo celular e aumentarem a imunogenicidade se tornando mais visível para o sistema imune. Crescentes evidências sugerem que o tratamento com DNMTis podem aumentar a sinalização imune nas células tumorais por meio do mecanismo de mimetismo viral bem como aumento da expressão de antígenos associados ao tumor, destacando assim, o seu potencial clínico na combinação com outras terapias. Enquanto os efeitos dos DNMTis têm sido na maior parte investigados nas células tumorais, os efeitos sobre o sistema imune ainda permanecem desconhecido, especialmente sobre baixas doses. Assim sendo, nosso objetivo foi investigar os efeitos da administração de agentes desmetilantes do DNA sobre a função efetora dos linfócitos T CD8 e seu papel frente a resposta antitumoral. Para isso, injetamos células tumorais CT26 na concentração de 5x105 por animal no dia 0. Cada grupo de animais foi tratado ou não com 5-AZA-CdR (0,5 mg/kg). Os camundongos foram tratados diariamente intraperitonealmente do dia 5 ao dia 9. Como controle adicional, em um grupo de animais, foi utilizado anticorpo neutralizante para depletar células T CD8. O tratamento diminuiu o tamanho tumoral, aumentou o infiltrado de células T CD8 e aumentou a capacidade de produção de citocinas por essas células. Em adição, todos esses efeitos foram abolidos na ausência das células T CD8. Em nossos ensaios de estimulação ex vivo e de imunização com adenovírus, o tratamento gerou aumento da ativação celular, aumento da produção de citocinas, diminuição da proliferação e aumento de morte celular. Por fim em nossos ensaios de citotoxicidade, o tratamento aumentou a capacidade citotóxica dessas células. Nossos resultados mostram que o tratamento in vivo com DNMTis em tumores de camundongos levou ao aumento da infiltração de células T CD8 e que a diminuição do tamanho tumoral é dependente da ação das células T CD8. Uma vez que observamos a capacidade efetoras das células T CD8, o tratamento levou ao aumento da produção de citocinas e aumento da indução de morte celular de células alvo. Em resumo, nossos resultados demonstram que o tratamento com DNMTis aumenta a resposta antitumoral e aumentam o potencial citotóxico das células T CD8. / Low doses of the DNA methylation inhibitor (DNMTis) on cancer cells has durable antitumorigenic effects by inducing cell cycle arrest and immune-modulatory properties that increases their visibility by the immune system. Growing evidence suggest that DNMTis can upregulate immune signaling in cancer cells through viral mimicry and upregulation of tumour associated antigens, highlighting clinical potential to combine with immunotherapy. While the effects of DNMTis have been mostly studied in cancer cells, their effects on the immune system remains vastly unknown specially at clinically relevant low doses. Therefore, we aimed to investigate the effect of in vivo administration of DNA demethylating inhibitor on the effector function of CD8+ T cells and antitumor immune response. We injected 5x105 CT26 cells at day 0 in three group of mice (n=5 mice per group). Each group of mice were either mock treated or treated with Decitabine (5-AZA-CdR or DAC, at a dosage of 0,5 mg/kg). The mice were treated intraperitoneally daily from days 5 to 9. As a control, in one group of mice, it was used neutralizing antibody to deplete CD8+ T cells. The treatment significantly decreased tumor burden and in the absence of CD8+ T cells this effect was abolished. The treatment led to increase in CD8+ T cells infiltration into the tumor microenvironment and enhance secretion of effector cytokines. In addition, all these effects were abolished in the lack of CD8 T cells. Furthermore, using ex vivo and adenovirus immunization, the treatment led to enhance of cellular activation, decrease of proliferation and enhance of cell death. In addition, using cytotoxic assays, the treatment enhanced the killing capability of these cells. to characterize the CD8 T cells, we used ex vivo stimulation protocols. Remarkably, our results show that treatment in vivo with DNMTi in established murine tumors model led to increase CD8+ T cell infiltration and the decrease in tumor burden is dependent of CD8 T cells. Once the effector capacities of these CD8+ T cells were assessed, DNMTi treatment increased their effector cytokine production and increased killing of target cells. Altogether, our results show that treatment of CD8+ T cells enhance antitumor immune response and their cytolytic potential.