RESUMEN
Checkpoint blockade immunotherapy has failed in pancreatic cancer and other poorly responsive tumor types in part due to inadequate T cell priming. Naive T cells can receive costimulation not only via CD28 but also through TNF superfamily receptors that signal via NF-κB. Antagonists of the ubiquitin ligases cellular inhibitor of apoptosis protein (cIAP)1/2, also called second mitochondria-derived activator of caspases (SMAC) mimetics, induce degradation of cIAP1/2 proteins, allowing for the accumulation of NIK and constitutive, ligand-independent activation of alternate NF-κB signaling that mimics costimulation in T cells. In tumor cells, cIAP1/2 antagonists can increase TNF production and TNF-mediated apoptosis; however, pancreatic cancer cells are resistant to cytokine-mediated apoptosis, even in the presence of cIAP1/2 antagonism. Dendritic cell activation is enhanced by cIAP1/2 antagonism in vitro, and intratumoral dendritic cells show higher expression of MHC class II in tumors from cIAP1/2 antagonism-treated mice. In this study, we use in vivo mouse models of syngeneic pancreatic cancer that generate endogenous T cell responses ranging from moderate to poor. Across multiple models, cIAP1/2 antagonism has pleiotropic beneficial effects on antitumor immunity, including direct effects on tumor-specific T cells leading to overall increased activation, increased control of tumor growth in vivo, synergy with multiple immunotherapy modalities, and immunologic memory. In contrast to checkpoint blockade, cIAP1/2 antagonism does not increase intratumoral T cell frequencies. Furthermore, we confirm our previous findings that even poorly immunogenic tumors with a paucity of T cells can experience T cell-dependent antitumor immunity, and we provide transcriptional clues into how these rare T cells coordinate downstream immune responses.
Asunto(s)
FN-kappa B , Neoplasias Pancreáticas , Ratones , Animales , FN-kappa B/metabolismo , Línea Celular Tumoral , Linfocitos T/metabolismo , Proteínas Inhibidoras de la Apoptosis , Apoptosis , InmunidadRESUMEN
We describe a de novo computational approach for designing proteins that recapitulate the binding sites of natural cytokines, but are otherwise unrelated in topology or amino acid sequence. We use this strategy to design mimics of the central immune cytokine interleukin-2 (IL-2) that bind to the IL-2 receptor ßγc heterodimer (IL-2Rßγc) but have no binding site for IL-2Rα (also called CD25) or IL-15Rα (also known as CD215). The designs are hyper-stable, bind human and mouse IL-2Rßγc with higher affinity than the natural cytokines, and elicit downstream cell signalling independently of IL-2Rα and IL-15Rα. Crystal structures of the optimized design neoleukin-2/15 (Neo-2/15), both alone and in complex with IL-2Rßγc, are very similar to the designed model. Neo-2/15 has superior therapeutic activity to IL-2 in mouse models of melanoma and colon cancer, with reduced toxicity and undetectable immunogenicity. Our strategy for building hyper-stable de novo mimetics could be applied generally to signalling proteins, enabling the creation of superior therapeutic candidates.
Asunto(s)
Diseño de Fármacos , Interleucina-15/inmunología , Interleucina-2/inmunología , Imitación Molecular , Receptores de Interleucina-2/agonistas , Receptores de Interleucina-2/inmunología , Secuencia de Aminoácidos , Animales , Sitios de Unión , Neoplasias del Colon/tratamiento farmacológico , Neoplasias del Colon/inmunología , Simulación por Computador , Cristalografía por Rayos X , Modelos Animales de Enfermedad , Humanos , Interleucina-15/uso terapéutico , Interleucina-2/uso terapéutico , Subunidad alfa del Receptor de Interleucina-2/inmunología , Subunidad alfa del Receptor de Interleucina-2/metabolismo , Melanoma/tratamiento farmacológico , Melanoma/inmunología , Ratones , Modelos Moleculares , Estabilidad Proteica , Receptores de Interleucina-2/metabolismo , Transducción de Señal/inmunologíaRESUMEN
Ipilimumab, a monoclonal antibody that recognizes cytotoxic T lymphocyte antigen (CTLA)-4, was the first approved "checkpoint"-blocking anticancer therapy. In mouse tumor models, the response to antibodies against CTLA-4 depends entirely on expression of the Fcγ receptor (FcγR), which may facilitate antibody-dependent cellular phagocytosis, but the contribution of simple CTLA-4 blockade remains unknown. To understand the role of CTLA-4 blockade in the complete absence of Fc-dependent functions, we developed H11, a high-affinity alpaca heavy chain-only antibody fragment (VHH) against CTLA-4. The VHH H11 lacks an Fc portion, binds monovalently to CTLA-4, and inhibits interactions between CTLA-4 and its ligand by occluding the ligand-binding motif on CTLA-4 as shown crystallographically. We used H11 to visualize CTLA-4 expression in vivo using whole-animal immuno-PET, finding that surface-accessible CTLA-4 is largely confined to the tumor microenvironment. Despite this, H11-mediated CTLA-4 blockade has minimal effects on antitumor responses. Installation of the murine IgG2a constant region on H11 dramatically enhances its antitumor response. Coadministration of the monovalent H11 VHH blocks the efficacy of a full-sized therapeutic antibody. We were thus able to demonstrate that CTLA-4-binding antibodies require an Fc domain for antitumor effect.
Asunto(s)
Antígeno CTLA-4/inmunología , Fragmentos Fc de Inmunoglobulinas/administración & dosificación , Fragmentos de Inmunoglobulinas/administración & dosificación , Neoplasias/terapia , Animales , Anticuerpos Monoclonales/administración & dosificación , Anticuerpos Monoclonales/química , Anticuerpos Monoclonales/inmunología , Antígeno CTLA-4/química , Línea Celular Tumoral , Modelos Animales de Enfermedad , Humanos , Fragmentos Fc de Inmunoglobulinas/química , Fragmentos Fc de Inmunoglobulinas/inmunología , Fragmentos de Inmunoglobulinas/química , Fragmentos de Inmunoglobulinas/inmunología , Inmunoglobulina G/administración & dosificación , Inmunoglobulina G/inmunología , Inmunoterapia , Ratones , Ratones Endogámicos C57BL , Neoplasias/inmunología , Dominios ProteicosRESUMEN
OBJECTIVE: The aims of this study were to assess first-year predoctoral dental students' knowledge, training, perceptions, and attitudes on obesity and to compare knowledge, perceptions, and attitudes before and after attending an educational presentation. METHODS: A presentation explaining the impact of obesity and its treatments on oral health was developed and presented to first-year predoctoral dental students (D1's) in the Doctor of Dental Medicine program. Before ("pre") and after ("post") the presentation, attending students were given the opportunity to take an anonymous and voluntary survey. Questions on the survey covered topics on knowledge, training, perceptions, and attitudes about obesity. RESULTS: Thirty-four of the 213 students in the course (response rate = 16.0%) attended the presentation and completed both the pre- and post-surveys. Most participating students reported receiving 0-5 h of education on obesity since starting their dental education. Regarding pre-survey answers to knowledge-based items, the percentage of students answering, "strongly agree" (representing the greatest knowledge level) ranged from 5.9% to 61.8% across items. Significant improvements from pre- to post-survey were observed for all knowledge-based items except a statement that obesity is associated with serious medical conditions. Significant improvements were also found for perception-based items asking about accommodations for patients and the importance of assessing dietary habits. Additionally, at post-survey, students reported significantly less discomfort asking about dietary habits and weight loss medications. CONCLUSION: Dental curricula policy makers and developers should consider the incorporation of obesity and its treatments, with nutrition experts teaching the content.
Asunto(s)
Actitud del Personal de Salud , Educación en Odontología , Conocimientos, Actitudes y Práctica en Salud , Obesidad , Estudiantes de Odontología , Humanos , Obesidad/terapia , Obesidad/psicología , Estudiantes de Odontología/psicología , Estudiantes de Odontología/estadística & datos numéricos , Encuestas y Cuestionarios , Femenino , MasculinoRESUMEN
Loss of major histocompatibility complex (MHC) class I and interferon-γ (IFN-γ) sensing are major causes of primary and acquired resistance to checkpoint blockade immunotherapy. Thus, additional treatment options are needed for tumors that lose expression of MHC class I. The cellular inhibitor of apoptosis proteins 1 and 2 (cIAP1/2) regulate classical and alternative nuclear factor κB (NF-κB) signaling. Induction of noncanonical NF-κB signaling with cIAP1/2 antagonists mimics costimulatory signaling, augmenting antitumor immunity. We show that induction of noncanonical NF-κB signaling induces T cell-dependent immune responses, even in ß2-microglobulin (ß2M)-deficient tumors, demonstrating that direct CD8 T cell recognition of tumor cell-expressed MHC class I is not required. Instead, T cell-produced lymphotoxin reprograms both mouse and human macrophages to be tumoricidal. In wild-type mice, but not mice incapable of antigen-specific T cell responses, cIAP1/2 antagonism reduces tumor burden by increasing phagocytosis of live tumor cells. Efficacy is augmented by combination with CD47 blockade. Thus, activation of noncanonical NF-κB stimulates a T cell-macrophage axis that curtails growth of tumors that are resistant to checkpoint blockade because of loss of MHC class I or IFN-γ sensing. These findings provide a potential mechanism for controlling checkpoint blockade refractory tumors.