RESUMO
Oncolytic virus (OV) therapy is an emerging approach with the potential to redefine treatment options across a range of cancer indications and in patients who remain resistant to existing standards of care, including immuno-oncology (IO) drugs. MEDI5395, a recombinant Newcastle disease virus (NDV), engineered to express granulocyte-macrophage colony-stimulating factor (GM-CSF), exhibits potent oncolytic activity. It was hypothesized that activation of immune cells by MEDI5395, coupled with its oncolytic activity, would enhance the priming of antitumor immunity. Using MEDI5395 and recombinant NDVs encoding fluorescent reporter genes, we demonstrated preferential virus uptake and non-productive infection in myeloid cells, including monocytes, macrophages, and dendritic cells (DCs). Infection resulted in immune-cell activation, with upregulation of cell surface activation markers (e.g., CD80, PD-L1, HLA-DR) and secretion of proinflammatory cytokines (IFN-α2a, IL-6, IL-8, TNF-α). Interestingly, in vitro M2-polarized macrophages were more permissive to virus infection than were M1-polarized macrophages. In a co-culture system, infected myeloid cells were effective virus vectors and mediated the transfer of infectious NDV particles to tumor cells, resulting in cell death. Furthermore, NDV-infected DCs stimulated greater proliferation of allogeneic T cells than uninfected DCs. Antigens released after NDV-induced tumor cell lysis were cross-presented by DCs and drove activation of tumor antigen-specific autologous T cells. MEDI5395 therefore exhibited potent immunostimulatory activity and an ability to enhance antigen-specific T-cell priming. This, coupled with its tumor-selective oncolytic capacity, underscores the promise of MEDI5395 as a multimodal therapeutic, with potential to both enhance current responding patient populations and elicit de novo responses in resistant patients.
Assuntos
Vírus da Doença de Newcastle/genética , Terapia Viral Oncolítica/métodos , Vírus Oncolíticos/genética , Linhagem Celular Tumoral , Vetores Genéticos , Humanos , Imunidade InataRESUMO
p63 is critical for epithelial development yet little is known about the transcriptional programmes it regulates. By characterising transcriptional changes and cellular effects following modulation of p63 expression, we have defined a vital role for p63 in cellular adhesion. Knockdown of p63 expression caused downregulation of cell adhesion-associated genes, cell detachment and anoikis in mammary epithelial cells and keratinocytes. Conversely, overexpression of the TAp63gamma or deltaNp63alpha isoforms of p63 upregulated cell adhesion molecules, increased cellular adhesion and conferred resistance to anoikis. Apoptosis induced by loss of p63 was rescued by signalling downstream of beta4 integrin. Our results implicate p63 as a key regulator of cellular adhesion and survival in basal cells of the mammary gland and other stratified epithelial tissues.
Assuntos
Células Epiteliais/citologia , Proteínas de Membrana/fisiologia , Anoikis , Adesão Celular/genética , Linhagem Celular Tumoral , Sobrevivência Celular/genética , Regulação da Expressão Gênica , Humanos , Queratinócitos/citologia , Proteínas de Membrana/genética , Isoformas de ProteínasRESUMO
The transition of ductal carcinoma in situ (DCIS) to invasive carcinoma is a poorly understood key event in breast tumor progression. Here, we analyzed the role of myoepithelial cells and fibroblasts in the progression of in situ carcinomas using a model of human DCIS and primary breast tumors. Progression to invasion was promoted by fibroblasts and inhibited by normal myoepithelial cells. Molecular profiles of isolated luminal epithelial and myoepithelial cells identified an intricate interaction network involving TGFbeta, Hedgehog, cell adhesion, and p63 required for myoepithelial cell differentiation, the elimination of which resulted in loss of myoepithelial cells and progression to invasion.
Assuntos
Neoplasias da Mama/patologia , Carcinoma Intraductal não Infiltrante/patologia , Neoplasias da Mama/genética , Carcinoma Intraductal não Infiltrante/genética , Adesão Celular , Metilação de DNA , Feminino , Regulação Neoplásica da Expressão Gênica , Homeostase , Humanos , Invasividade Neoplásica , Análise de Sequência com Séries de Oligonucleotídeos , Polimorfismo de Nucleotídeo Único , Fator de Crescimento Transformador beta/genética , Proteína Supressora de Tumor p53/genéticaRESUMO
The development of stratified epithelia and their derivatives is a complex process requiring a multifaceted transcriptional program. p63, the p53-related transcription factor, is fundamental to this process. However, the underlying mechanisms by which p63 exerts its influence on stratified epithelial development and integrity remain elusive. Recent work from our laboratories has demonstrated that p63 mediates its effects on stratified epithelial function at least in part via its ability to regulate multiple aspects of epithelial cellular adhesion and survival. The identification of cell-cell and cell-matrix adhesion subprograms downstream of p63 provides an initial understanding of p63's role in epithelial development, integrity and homeostasis.