On-target restoration of a split T cell-engaging antibody for precision immunotherapy

T cell-engaging immunotherapies are changing the landscape of current cancer care. However, suitable target antigens are scarce, restricting these strategies to very few tumor types. Here, we report on a T cell-engaging antibody derivative that comes in two complementary halves and addresses antigen combinations instead of single molecules. Each half, now coined hemibody, contains an antigen-specific single-chain variable fragment (scFv) fused to either the variable light (V-L) or variable heavy (V-H) chain domain of an anti-CD3 antibody. When the two hemibodies simultaneously bind their respective antigens on a single cell, they align and reconstitute the original CD3-binding site to engage T cells. Employing preclinical models for aggressive leukemia and breast cancer, we show that by the combinatorial nature of this approach, T lymphocytes exclusively eliminate dual antigen-positive cells while sparing single positive bystanders. This allows for precision targeting of cancers not amenable to current immunotherapies

Das immunevasive Potential von MYC im Pankreaskarzinom

Pancreatic ductal adenocarcinoma (PDAC) is predominantly driven by mutations in KRAS and TP53. However, PDAC tumors display deregulated levels of MYC and are a paradigm example for MYC-driven and -addicted tumors. For many years MYC was described as a transcription factor that regulates a pleiotropic number of genes to drive proliferation. Recent work sheds a different light on MYC biology. First, changes in gene expression that come along with the activation of MYC are mild and MYC seems to act more as a factor that reduces stress and increases resilience towards challenges during transcription. Second, MYC is a strong driver of immune evasion in different entities. In this study we depleted MYC in murine PDAC cells and revealed the immune dependent regression of tumors in an orthotope transplant model, as well as the activation of the innate immune system using global expression analysis, immunoblotting and fCLIP. These experiments revealed that endogenous double-stranded RNA is binding as a viral mimicry to Toll-like receptor 3, causing activation of TBK1 and downstream activation of a proimmunogenic transcription program. The regression of tumors upon depletion of MYC is dependent on this pathway since the knockout of TBK1 prevents regression of tumors after depletion of MYC. We can summarize this study in three main findings: First, the dominant and most important function of MYC in tumors is not to drive proliferation but to promote immune evasion and prevent immune-dependent regression of tumors. Second, cells monitor defects or delay in splicing and RNA processing and activate the immune system to clear cells that face problems with co-transcriptional processing. Third, MYC suppresses the activation of the cell-intrinsic innate immune system and shields highly proliferating cells from the recognition by the immune system. To translate this into a therapeutically approach, we replaced the shRNA mediated depletion of MYC by treatment with cardiac glycosides. Upon treatment with cardiac glycosides tumor cells reduce uptake of nutrients, causing a downregulation of MYC translation, inhibition of proliferation, glycolysis and lactate secretion. Lactate is a major reason for immune evasion in solid tumors since it dampens, amongst others, cytotoxic T cells and promotes regulatory T cells. Treatment of mice with cardiac glycosides causes a complete and immune-dependent remission of PDAC tumors in vivo, pointing out that cardiac glycosides have strong proimmunogenic, anti-cancer effects. More detailed analyses will be needed to dissect the full mechanism how cardiac glycosides act on MYC translation and immune evasion in PDAC tumors.Pankreaskarzinome entwickeln sich in den meisten Fällen durch die Mutation von KRAS und TP53. Nichtsdestotrotz weisen Pankreaskarzinome sehr hohe, deregulierte Level des MYC Proteins auf und sind exemplarisch für Tumore, deren Wachstum abhängig von MYC ist. Für lange Zeit wurde MYC als Transkriptionsfaktor beschrieben, der vor allem Gene aktiviert, die für die Proliferation von Zellen notwendig sind. Die jüngste Forschung wirft jedoch ein anderes Bild auf die Biologie von MYC. Zum einen sind die transkriptionellen Veränderung nach Aktivierung von MYC mild und vieles deutet darauf hin, dass die Funktion von MYC zum einen in der Reduktion von transkriptionellem Stress liegt. Zum anderen verhindert MYC die Erkennung von Tumorzellen durch das Immunsystem. In dieser Studie wurde MYC in murinen Pankreaskarzinomzellen depletiert und die immunabhängige Regression der Tumore in einem orthotopen Transplantationsmodel untersucht. Die Aktivierung der zell-intrinsischen Immunantwort wurde mittels globaler Expressionsanalyse, Immunoblots und fCLIP Experimenten untersucht. Diese Experimente haben gezeigt, dass endogene doppel-strängige RNAs als virales Mimikry an Toll-like Rezeptor 3 binden, worauf TBK1 phosphoryliert und ein pro-immunogenes Transkriptionsprogram aktiviert wird. Die Deletion von TBK1 konnte beweisen werden, dass die Regression der Tumore von diesem Signalweg abhängig ist. Die Ergebnisse der Studie lassen sich in drei zentrale Erkenntnisse zusammenfassen: Erstens ist die prä-dominante Funktion von MYC in Tumoren in vivo nicht die Proliferation der Zellen zu fördern, sondern zu verhindern, dass der Tumor vom Immunsystem erkannt und bekämpft wird. Zweitens überwachen Zellen das richtige Prozessieren von RNA und aktivieren bei Bedarf das Immunsystem um defekte Zellen zu entfernen. Drittens unterdrückt MYC die Aktivierung dieses zell-intrinsischen Signalweges und schirmt den Tumor dadurch vom Immunsystem ab. Um diese Ergebnisse in einen translationalen Ansatz zu überführen, haben wir die shRNA vermittelte Depletion von MYC durch die Behandlung mit Herzglykosiden ersetzt, die die Aufnahme von Nährstoffen reduzieren, Proliferation einschränken, die Translation des MYC Proteins stoppen und Glykolyse und Laktatsekretion herunterfahren. Laktat hat starke immunsuppressive Eigenschaften in soliden Tumoren, da es zytotoxische T Zellen erschöpft und regulatorische T Zellen aktiviert. Die Behandlung von Mäusen mit Herzglykosiden führte in Abhängigkeit vom Immunsystem zur kompletten Remission der Tumore, auch wenn noch weitere Forschung notwendig ist, um die exakten Zusammenhänge besser zu verstehen

Nurses’ voice: the role of hierarchy and leadership

Voicing concerns and suggestions is crucial for preventing medical errors and improving patient safety. Research suggests that hierarchy in health-care teams impair open communication. Hierarchy, however, can vary with changing team composition, particularly during acute care situations where more senior persons join the team later on. The purpose of this study is to investigate how changes in hierarchy and leadership were associated with nurses’ voice frequency and nurses’ time to voice during simulated acute care situations

PAF1c links S-phase progression to immune evasion and MYC function in pancreatic carcinoma

Abstract In pancreatic ductal adenocarcinoma (PDAC), endogenous MYC is required for S-phase progression and escape from immune surveillance. Here we show that MYC in PDAC cells is needed for the recruitment of the PAF1c transcription elongation complex to RNA polymerase and that depletion of CTR9, a PAF1c subunit, enables long-term survival of PDAC-bearing mice. PAF1c is largely dispensable for normal proliferation and regulation of MYC target genes. Instead, PAF1c limits DNA damage associated with S-phase progression by being essential for the expression of long genes involved in replication and DNA repair. Surprisingly, the survival benefit conferred by CTR9 depletion is not due to DNA damage, but to T-cell activation and restoration of immune surveillance. This is because CTR9 depletion releases RNA polymerase and elongation factors from the body of long genes and promotes the transcription of short genes, including MHC class I genes. The data argue that functionally distinct gene sets compete for elongation factors and directly link MYC-driven S-phase progression to tumor immune evasion

Sagebrush Control: At What Canopy Cover Is It Economically Justified?

We determine the economic threshold level for big sagebrush control based on 18 yr of forage-response data from an experiment conducted in Carbon County, Wyoming. We analyze the impacts of climatic variables and treatment site characteristics, such as sagebrush abundance levels, precipitation, and understory composition, on forage response and threshold level. We find that sagebrush canopy cover levels, April precipitation, May soil moisture, and understory composition are statistically significant factors in explaining forage response to sagebrush treatment. Forage yield across treated and untreated plots for 10 canopy cover levels, ranging from 4% to 40%, are analyzed via panel data regression techniques. We further investigate the impact of variability in precipitation and understory characteristics on economic outcomes of sagebrush control by analyzing three scenarios. Scenario 1 uses actual forage response data that include all variability from precipitation and understory composition. Scenario 2 uses regression-predicted yields across plots assuming average precipitation and soil moisture conditions. Scenario 3 uses regression-predicted yields assuming average precipitation, soil moisture, and understory characteristics across plots. Net present values based on value of grazing (for estimated yield differences between treated and untreated plots assuming 50% forage utilization) compared to treatment cost across sagebrush cover levels are estimated across these three scenarios. Results indicate that the economic threshold level of sagebrush infestation for the study period was between 8% and 24% for the analyzed scenarios. This indicates variability in precipitation and understory composition impact forage response and the resulting economics of sagebrush control. We conclude that range managers should consider potential control site characteristics and long-range weather forecasts when contemplating sagebrush control./Determinamos el nivel del umbral económico para el control de la artemisa basados en 18 años de datos sobre la respuesta del forraje en un experimento realizado en Carbon County, Wyoming. Analizamos el impacto de variables climáticas y características del sitio tales como niveles de abundancia de artemisa, precipitación y la composición abajo del dosel en la respuesta del forraje y el nivel de umbral. Encontramos que los niveles de cobertura aérea de la artemisa, la precipitación de abril, la humedad del suelo de mayo y la composición son factores estadísticamente significantes para explicar la respuesta del forraje en el tratamiento de la artemisa. El rendimiento de forraje a lo largo de las parcelas tratadas y no tratadas para diez niveles de cubierta aérea fluctuaron del 4% al 40% son analizados por medio de técnicas de regresión de datos panel. Además investigamos el impacto de la variabilidad en precipitación y características debajo del dosel en los resultados económicos del control de la artemisa analizando tres escenarios. En el escenario uno, se usaron los datos de la respuesta actual del forraje la cual incluye toda la variabilidad de la precipitación y composición de abajo del dosel. El escenario dos, usa rendimientos predichos de regresión a lo largo de las parcelas asumiendo precipitación promedio y condiciones de humedad del suelo. El escenario tres usa rendimientos predichos de regresión asumiendo precipitación promedio, humedad del suelo y características de abajo del dosel a través de las parcelas. Valores presentes netos basados en el valor del pastoreo (estimados de las diferencias entre los rendimientos de las parcelas tratadas y no tratadas asumiendo un 50% de utilización del forraje) comparado con el costo del tratamiento a través de los niveles de cobertura de la artemisa son estimados a través de estos tres escenarios. Los resultados indican que el nivel del umbral económico de infestación de artemisa para el periodo de estudio fue entre 8% y 24% de los escenarios analizados. Esto indica que la variabilidad en precipitación y composición abajo del dosel impacta la respuesta del forraje resultando en el control económico de la artemisa. Concluimos que manejadores de pastizales deben considerar las características potenciales de control en el sitio y rangos amplios de pronósticos de tiempo cuando consideren el control de la artemisa.The Rangeland Ecology & Management archives are made available by the Society for Range Management and the University of Arizona Libraries. Contact [email protected] for further information.Migrated from OJS platform August 202