RESUMEN
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.
Asunto(s)
Humanos , Terapia Genética/métodos , Sistemas de Lectura/genética , Interferón beta/uso terapéutico , Técnicas de Transferencia de Gen , Inmunoterapia/métodos , Neoplasias/terapia , Muerte Celular/genética , Inhibidor p16 de la Quinasa Dependiente de Ciclina/genética , Proteína p14ARF Supresora de Tumor/genética , Neoplasias/inmunologíaRESUMEN
O câncer de mama é uma doença extremamente heterogênea compreendendo diferentes subtipos moleculares que resultam em evoluções clínicas e condutas terapêuticas distintas. A maior gravidade desta patologia está associada a sua capacidade de formação de metástases Mudanças no padrão de expressão gênica têm sido associadas à manifestação do fenótipo metastático. Neste trabalho, utilizamos microarranjos de tecido (TMAs) para investigar a expressão de 8 biomarcadores candidatos (CIP4, PPIL1, ITGAV, AKAP14, MICA, FXYD1, ARPC3, ABG1) e avaliar seu potencial prognóstico em pacientes com carcinoma ductal invasivo da mama. Destes, ARPC3 PPIL1 e CIP4 mostraram associações estatisticamente significativas com a sobrevida câncer específica e/ou a probabilidade de desenvolvimento de metástases. Determinamos que a expressão aumentada de CIP4 nos tumores está associada a maior probabilidade de desenvolvimento de metástases. CIP4 é uma proteína adaptadora descrita na literatura como moduladora de migração e invasão celular e portanto selecionamos este candidato para caracterização funcional detalhada. Observamos que a expressão de CIP4 encontra-se aumentada em linhagens tumorais com características invasivas. A partir do silenciamento estável e regulado de CIP4 na linhagem metastática MDA-MB-231, determinamos que CIP4 modula positivamente a ativação de MAPK-p38 e a expressão de MMP2 , sugerindo que CIP4 participe em vias de sinalização importantes para a transição epitélio-mesenquima (EMT). O silenciamento de CIP4 resultou em uma redução de aproximadamente 50% da capacidade migratória e invasiva das células tumorais in vitro , e na diminuição da formação de metástases pulmonares in vivo. Coletivamente, nossos resultados indicam que CIP4 tem potencial como marcador de prognóstico assim como um possível alvo terapêutico no controle da disseminação de metástases nos tumores da mama
Breast cancer is an extremely heterogeneous disease comprising different molecular subtypes that result in different clinical outcomes and therapeutic procedures. The severity of this disease is mainly associated with its ability to produce metastasis. Changes in gene expression profile have been associated with the manifestation of the metastatic phenotype. In this study, we used tissue microarrays (TMAs) to investigate the expression of 8 candidate biomarkers (CIP4, PPIL1, ITGAV, AKAP14, MICA, FXYD1, ARPC3 e ABG1) and to evaluate their prognostic potential in patients with invasive ductal breast carcinoma. Among these, ARPC3, PPIL1 and CIP4 showed statistically significant associations with cancer specific survival and/or the patient's probability to develop metastasis. We found that increased expression of CIP4 in tumors is associated with a higher probability of developing metastasis. CIP4 is an adaptor protein described in the literature as a modulator of cell migration and invasion and therefore we selected this candidate for detailed functional characterization. We observed that CIP4 expression is increased in tumor cell lines with invasive characteristics. Following the stable and regulated knockdown of CIP4 in the metastatic line MDA-MB-231, we determined that it modulates positively the activation of MAPK-p38 and the expression of MMP2, suggesting that CIP4 participates in important signaling pathways required for the epithelial mesenchymal transition (EMT). CIP4 silencing resulted in an approximate 50% reduction of the migratory and invasive capacity of tumor cells in vitro and decreased the generation of lung metastases in vivo. Collectively, our results indicate that CIP4 has potential as a prognostic marker as well as a potential therapeutic target to control the metastatic dissemination of breast tumors