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SUMMARY We present a semi-analytical method and expressions for computing the displacements, strains and stress due to localized (e.g. faulting) and distributed (volumetric) inelastic deformation in heterogeneous elastic full- and half-spaces. Variations in elastic properties are treated as piecewise-constant homogeneous subregions as in orthodox multiregion approaches. The deformation in the subregions is solved by matching the interface traction and displacement conditions for contrasting elastic parameters. We show equivalence between the integral equation convolving boundary traction and those convolving displacement discontinuities and volumetric inelastic strain in the representation theorem for a bounded volume. This equivalence allows us to express the deformation fields in the half-/full-space which comprises those subregions by using virtual fault displacement elements or volumetric eigenstrain elements, the integral kernels of which have known analytic forms for finite sources in homogeneous volumes. We include computer programs that implement our method with known analytic solutions of homogeneous volumes free of major singular points. We provide an extension to the existing toolkit available for the observational and theoretical analyses of deformation fields allowing users to model heterogeneous geological structures, with a number of primary geophysical applications, including earthquake and volcano deformation, where variations in elastic parameters may present a substantial contribution to the observed deformation.

Faults and fractures are a critical store and pathway for groundwater in Ireland’s limestone bedrock aquifers either directly as conductive structures or indirectly as the locus for the development of karst conduits. From the quantitative analysis of post-Devonian faults and fractures in a range of lithological sequences, this report describes the principal characteristics of Cenozoic strike-slip faults and joints, the youngest and the most intrinsically conductive fractures within Irish bedrock. Analysis of these structures in more than 120 outcrop, quarry, mine and cave locations in a range of bedrock types, provides a basis for: (1) definition of quantitative models for their depth dependency, lithological control, scaling systematics and links to preexisting structure, (2) conceptualisation of their impact on groundwater behaviour, and (3) estimation of groundwater flow parameters. The quantitative models provide constraints on fracture-controlled flow connectivity. Commonly observed decreases in sustainable flows and water strike interceptions with depth are attributed to increasing confinement and decreasing fracture connectivity and dissolution. Faults and joints have quite different end member geometries, with faults having strongly heterogeneous scale-independent properties and joints more often showing scale-dependent stratabound properties. The highest and most sustainable groundwater flows are usually associated with the complexity of structure of Cenozoic faults and of preexisting Carboniferous structures (on which conductive fracturing localises), enhanced by karstification and strongly jointed limestone bedrock particularly in the near-surface. Increased groundwater flow is promoted within bedded, rather than massive (i.e. unbedded), limestone sequences, characterised by bedding-parallel fractures and karst connecting otherwise subvertical fractures and subvertical wells.

A hydrogeological conceptual model of the source, circulation pathways and temporal variation of a low-enthalpy thermal spring in a fractured limestone setting is derived from a multidisciplinary approach. St. Gorman's Well is a thermal spring in east-central Ireland with a complex and variable temperature profile (maximum of 21.8 °C). Geophysical data from a three-dimensional(3D)audio-magnetotelluric(AMT) survey are combined with time-lapse hydrogeological data and information from a previously published hydrochemical analysis to investigate the operation of this intriguing hydrothermal system. Hydrochemical analysis and time-lapse measurements suggest that the thermal waters flow within the fractured limestones of the Carboniferous Dublin Basin at all times but display variability in discharge and temperature. The 3D electrical resistivity model of the subsurface revealed two prominent structures: (1) a NW-aligned faulted contact between two limestone lithologies; and (2) a dissolutionally enhanced, N-aligned, fault of probable Cenozoic age. The intersection of these two structures, which has allowed for karstification of the limestone bedrock, has created conduits facilitating the operation of relatively deep hydrothermal circulation (likely estimated depths between 240 and 1,000 m) within the limestone succession of the Dublin Basin. The results of this study support a hypothesis that the maximum temperature and simultaneous increased discharge observed at St. Gorman's Well each winter is the result of rapid infiltration, heating and recirculation of meteoric waters within a structurally controlled hydrothermal circulation system.The online version contains supplementary material available at 10.1007/s10040-021-02393-1.Un modèle conceptuel hydrogéologique de la source, des voies de circulation et de la variation temporelle d’une source thermale à faible enthalpie dans un contexte de calcaire fracturé est dérivé d’une approche multidisciplinaire. St. Gorman’s Well est une source thermale du centre-est de l’Irlande avec un profil de température complexe et variable (maximum de 21.8 °C). Les données géophysiques d’un levé audio-magnétotellurique (AMT) en trois dimensions (3D) sont combinées avec des données hydrogéologiques à intervalles de temps et des informations provenant d’une analyse hydrochimique publiée précédemment pour étudier le fonctionnement de cet intrigant système hydrothermal. L’analyse hydrochimique et les mesures à différentes périodes suggèrent que les eaux thermales s’écoulent à tout moment dans les calcaires fracturés du bassin carbonifère de Dublin, mais présentent une variabilité de débit et de température. Le modèle de résistivité électrique 3D du sous-sol a révélé deux structures importantes: (1) un contact faillé orienté NW entre deux lithologies calcaires; et (2) une faille alignée au Nord, améliorée par dissolution, d’âge cénozoïque probable. L’intersection de ces deux structures, qui a permis la karstification du socle calcaire, a créé des conduits facilitant le fonctionnement d’une circulation hydrothermale relativement profonde (profondeurs estimées vraisemblablement entre 240 et 1,000 m) au sein de la succession calcaire du bassin de Dublin. Les résultats de cette étude appuient l’hypothèse selon laquelle la température maximale et l’augmentation simultanée du débit observés à St. Gorman’s Well chaque hiver sont le résultat d’une infiltration, d’un réchauffement et d’une recirculation rapides des eaux météoriques dans un système de circulation hydrothermale structurellement contrôlé.Se deriva un modelo conceptual hidrogeológico de la fuente, las vías de circulación y la variación temporal de un manantial termal de baja entalpía en un entorno de caliza fracturada a partir de un enfoque multidisciplinar. Gorman’s Well es un manantial termal en el centro-este de Irlanda con un perfil de temperatura complejo y variable (máximo de 21.8 °C). Los datos geofísicos de un estudio audio-magnetotelúrico (AMT) tridimensional (3D) se combinan con los datos hidrogeológicos de un lapso de tiempo y la información de un análisis hidroquímico previamente publicado para investigar el funcionamiento de este intrigante sistema hidrotermal. El análisis hidroquímico y las mediciones a intervalos de tiempo sugieren que las aguas termales fluyen dentro de las calizas fracturadas de la cuenca carbonífera de Dublín en todo momento, pero muestran variabilidad en la descarga y la temperatura. El modelo de resistividad eléctrica tridimensional del subsuelo reveló dos estructuras prominentes: (1) un contacto de falla alineado al NW entre dos litologías calcáreas; y (2) una falla de disolución incrementada, alineada al N, de probable edad cenozoica. La intersección de estas dos estructuras, que ha permitido la karstificación del lecho rocoso calcáreo, ha creado conductos que facilitan el funcionamiento de una circulación hidrotermal relativamente profunda (probablemente a profundidades estimadas entre 240 y 1,000 m) dentro de la sucesión calcárea de la cuenca de Dublín. Los resultados de este estudio apoyan la hipótesis de que la temperatura máxima y el aumento simultáneo de la descarga observados en St. Gorman’s Well cada invierno son el resultado de una rápida infiltración, calentamiento y recirculación de aguas meteóricas dentro de un sistema de circulación hidrotermal estructuralmente controlado.通过多学科方法, 建立了裂缝型石灰岩环境中低焓温泉的来源、循环路径和时间变化的水文地质概念模型。圣戈尔曼井是爱尔兰中东部的温泉, 温度分布复杂多变 (最高21.8 °C)。将三维 (3D) 音频-大地电磁 (AMT) 调查的地球物理数据与延时水文地质数据和先前发布的水化学分析信息相结合, 调查了有趣的热液系统运行情况。水化学分析和延时测量表明, 热水一直在石炭系都柏林盆地的裂缝型石灰岩内流动, 但流量和温度有变异性。地下的 3D 电阻率模型揭示了两个突出的结构:(1) 两个石灰岩岩性之间的 NW 向断层接触; 和 (2) 可能新生代的溶蚀增强的和N向的断层。两个结构的交叉点使石灰岩基岩发生岩溶作用, 形成了通道, 促进了都柏林盆地石灰岩序列内相对较深的热液循环 (估计深度可能在 240 至 1,000 m 之间)作用。通过这项研究发现每年冬天在圣戈尔曼井观察到的最高温度和同时增加的排泄量是结构控制的热液循环系统中大气水快速入渗、加热和再循环的结果。.Um modelo hidrogeológico conceitual da fonte, vias de circulação e variação temporal de uma fonte termal de baixa entalpia em um ambiente de calcário fraturado é derivado de uma abordagem multidisciplinar. O poço de St. Gorman é uma fonte termal no centro-leste da Irlanda com um perfil de temperatura complexo e variável (máximo de 21.8°C). Os dados geofísicos de uma pesquisa áudio-magnetotelúrica (AMT) tridimensional (3D) são combinados com dados hidrogeológicos em intervalos de tempo e informações de uma análise hidroquimica publicada anteriormente, para investigar a operação deste intrigante sistema hidrotérmico. A análise hidroquimica e as medições em intervalos de tempo sugerem que as águas termais fluem de dentro dos calcários fraturados da Bacia Carbonifera de Dublin o tempo todo, mas exibem variabilidade na descarga e na temperatura. O modelo de resistividade elétrica 3D da subsuperfície revelou duas estruturas proeminentes: (1) um contato defeituoso alinhado a NO entre duas litologias de calcário; e (2) uma falha dissolucionalmente aumentada, alinhada a N, de provável idade Cenozóica. A intersecção dessas duas estruturas, que permitiu a carstificação da rocha calcária, criou condutos que facilitam a operação de circulação hidrotérmica relativamente profunda (profundidade estimada entre 240 e 1,000 m) dentro da sucessão de calcário da Bacia Dublin. Os resultados desse estudo suportam a hipótese de que a temperatura máxima e o aumento simultâneo da descarga observada no poço de St. Gorman a cada inverno é o resultado da rápida infiltração, aquecimento e recirculação de águas meteóricas dentro de um sistema de circulação hidrotérmica estruturalmente controlado.

PurposeExposures early in life, beginning in utero, have long-term impacts on mental and physical health. The ECHO prenatal and early childhood pathways to health consortium (ECHO-PATHWAYS) was established to examine the independent and combined impact of pregnancy and childhood chemical exposures and psychosocial stressors on child neurodevelopment and airway health, as well as the placental mechanisms underlying these associations.ParticipantsThe ECHO-PATHWAYS consortium harmonises extant data from 2684 mother–child dyads in three pregnancy cohort studies (CANDLE [Conditions Affecting Neurocognitive Development and Learning in Early Childhood], TIDES [The Infant Development and Environment Study] and GAPPS [Global Alliance to Prevent Prematurity and Stillbirth]) and collects prospective data under a unified protocol. Study participants are socioeconomically diverse and include a large proportion of Black families (38% Black and 51% White), often under-represented in research. Children are currently 5–15 years old. New data collection includes multimodal assessments of primary outcomes (airway health and neurodevelopment) and exposures (air pollution, phthalates and psychosocial stress) as well as rich covariate characterisation. ECHO-PATHWAYS is compiling extant and new biospecimens in a central biorepository and generating the largest placental transcriptomics data set to date (N=1083).Findings to dateEarly analyses demonstrate adverse associations of prenatal exposure to air pollution, phthalates and maternal stress with early childhood airway outcomes and neurodevelopment. Placental transcriptomics work suggests that phthalate exposure alters placental gene expression, pointing to mechanistic pathways for the developmental toxicity of phthalates. We also observe associations between prenatal maternal stress and placental corticotropin releasing hormone, a marker of hormonal activation during pregnancy relevant for child health. Other publications describe novel methods for examining exposure mixtures and the development of a national spatiotemporal model of ambient outdoor air pollution.Future plansThe first wave of data from the unified protocol (child age 8–9) is nearly complete. Future work will leverage these data to examine the combined impact of early life social and chemical exposures on middle childhood health outcomes and underlying placental mechanisms.

Despite estimates of climate target’s remaining carbon budget (RCB) informing critical policy, most climate projections do not account for peatland ecosystem processes, including the contribution of current or future peatland wildfires to global carbon emissions. Here, we provide the first estimate of the carbon emissions associated with non-permafrost northern peatlands that specifically includes both wildfire combustion and post-fire carbon dynamics. The inclusion of wildfire reduced the carbon sink from -59.5 ± 32.2 to -17.9 ± 45.7 g C m-2 yr-1 in natural (pristine) peatlands while degraded peatlands represented a consistent source of carbon (218.6 ± 48.1 g C m-2 yr-1). We find that small increases in average burn severity or peatland area burned (from 0.5 to 0.8 % yr-1) could tip northern peatlands from a net carbon sink to a net source, illustrating the critical importance of peatland wildfire emissions and their accounting to the RCB for global climate targets.

OBJECTIVE: To evaluate episode‐based payments for upper respiratory tract infections (URI) and perinatal care in Arkansas's Medicaid population. STUDY SETTING: Upper respiratory infection and perinatal episodes among Medicaid‐covered individuals in Arkansas and comparison states from fiscal year (FY) 2011 to 2014. STUDY DESIGN: Cross‐sectional observational analysis using a difference‐in‐difference design to examine outcomes associated with URI and perinatal episodes of care (EOC) from 2011 to 2014. Key dependent variables include antibiotic use, emergency department visits, physician visits, hospitalizations, readmission, and preventive screenings. DATA COLLECTION: Claims data from the Medicaid Analytic Extract for Arkansas, Mississippi, and Missouri from 2010 to 2014 with supplemental county‐level data from the Area Health Resource File (AHRF). PRINCIPAL FINDINGS: The URI EOC reduced the probability of antibiotic use (marginal effect [ME] = −1.8, 90% CI: −2.2, −1.4), physician visits (ME = 0.6, 90% CI: −0.8, −0.4), improved the probability of strep tests for children diagnosed with pharyngitis (ME = 9.4, 90% CI: 8.5, 10.3), but also increased the probability of an emergency department (ED) visit (ME = 0.1, 90% CI: 0.1, 0.2), relative to the comparison group. For perinatal EOCs, we found a reduced probability of an ED visit during pregnancy (ME = 0.1, 90% CI: −0.2, −0.0), an increased probability of screening for HIV (ME = 6.2, 90% CI: 4.0, 8.5), chlamydia (ME = 9.5, 90% CI: 7.2, 11.8), and group B strep‐test (ME = 2.6, 90% CI: 0.5, 4.6), relative to the comparison group. Predelivery and postpartum hospitalizations also increased (ME = 1.2, 90% CI: 0.4, 2.0; ME = 0.4, 90% CI: 0.0, 0.8, respectively), relative to the comparison group. CONCLUSION: Upper respiratory infection and perinatal EOCs for Arkansas Medicaid beneficiaries produced mixed results. Aligning shared savings with quality metrics and cost‐thresholds may help achieve quality targets and disincentivize over utilization within the EOC, but may also have unintended consequences.

Comunicació presentada a 6th International Congress on Information and Communication Technology (ICICT 2021), celebrat del 25 al 26 de febrer de 2021 de manera virtual. Autism is a neurodevelopmental disorder characterized by deficits in social communication and repetitive patterns of behavior. Individuals affected by Autism Spectrum Disorder (ASD) may face overwhelming sensory hypersensitivities that hamper their everyday life. In order to promote awareness about neurodiversity among the neurotypical population, we have developed an interactive virtual reality simulation to experience the oversensory stimulation that an individual with autism spectrum disorder may experience in a natural environment. In this experience, we project the user in a first-person perspective in a classroom where a teacher is presenting a lecture. As the user explores the classroom and attends the lecture, he/she is confronted with sensory distortions which are commonly experienced by persons with ASD. We provide the users with a virtual reality headset with motion tracking, two wireless controllers for interaction, and a wristband for physiological data acquisition to create a closed feedback loop. This wearable device measures blood volume pulse (BVP) and electrodermal activity (EDA), which we use to perform online estimations of the arousal levels of users as they respond to the virtual stimuli. We use this information to modulate the intensity of auditory and visual stimuli simulating a vicious cycle in which increased arousal translates into increased oversensory stimulation. Here, we present the architecture and technical implementation of this system. This research received funding from H2020-EU, ID: 787061 and ERC (PoC) H2020-EU, ID: 840052.

We present the findings of "The Alzheimer's Disease Prediction Of Longitudinal Evolution" (TADPOLE) Challenge, which compared the performance of 92 algorithms from 33 international teams at predicting the future trajectory of 219 individuals at risk of Alzheimer's disease. Challenge participants were required to make a prediction, for each month of a 5-year future time period, of three key outcomes: clinical diagnosis, Alzheimer's Disease Assessment Scale Cognitive Subdomain (ADAS-Cog13), and total volume of the ventricles. The methods used by challenge participants included multivariate linear regression, machine learning methods such as support vector machines and deep neural networks, as well as disease progression models. No single submission was best at predicting all three outcomes. For clinical diagnosis and ventricle volume prediction, the best algorithms strongly outperform simple baselines in predictive ability. However, for ADAS-Cog13 no single submitted prediction method was significantly better than random guesswork. Two ensemble methods based on taking the mean and median over all predictions, obtained top scores on almost all tasks. Better than average performance at diagnosis prediction was generally associated with the additional inclusion of features from cerebrospinal fluid (CSF) samples and diffusion tensor imaging (DTI). On the other hand, better performance at ventricle volume prediction was associated with inclusion of summary statistics, such as the slope or maxima/minima of biomarkers. TADPOLE's unique results suggest that current prediction algorithms provide sufficient accuracy to exploit biomarkers related to clinical diagnosis and ventricle volume, for cohort refinement in clinical trials for Alzheimer's disease. However, results call into question the usage of cognitive test scores for patient selection and as a primary endpoint in clinical trials., Comment: Presents final results of the TADPOLE competition. 60 pages, 7 tables, 14 figures

Parasites can alter a wide range of host behaviors resulting in changes in organismal interactions and ecosystem processes. One of the most important behaviors that controls food web dynamics is herbivore grazing because an alteration in grazing behavior leads to changes in trophic dynamics and ecosystem processes by changing the abundance and diversity of primary producers. To test whether parasite load can alter host grazing levels and choices, feeding trials were conducted using the keystone species, the rusty crayfish Faxonius rusticus (Girard, 1852), grazing on a selection of macrophyte species. The rusty crayfish is a keystone species because its grazing significantly alters the abundance of macrophytes in freshwater ecosystems. We used a total of 165 wild-caught, naturally-infected crayfish individuals with a wide range of parasite loads by species of the digenetic trematode Microphallus Ward, 1901. Crayfish were presented with 1 g each of the macrophytes Elodea canadensis (Michaux), Ceratophyllum demersum (L), Chara sp., and Potamogeton richardsonii (A. Benn.) in a 23-hr foraging assay. Subsequently, crayfish were dissected, and parasite loads were calculated. Mixed models were then utilized to determine how parasite load affected consumption. As infection of Microphallus increased in the crayfish hepatopancreas, consumption of all four macrophytes significantly decreased. Melanization of Microphallus spp. within the hepatopancreas, the immune response to. infection, did not significantly reduce crayfish macrophyte consumption. These results indicate that macrophyte consumption in the crayfish was affected by Microphallus. This impact on crayfish grazing could alter macrophyte abundances in aquatic ecosystems. Because of the many ecosystem functions macrophytes play, an alteration in their abundances could lead to community-level ramifications by impacting nutrient flow and organismal abundances in aquatic ecosystems.

Background Tebotelimab, also known as MGD013, is an investigational, Fc bearing bispecific tetravalent DART molecule designed to bind PD-1 and LAG-3 and sustain/restore the function of exhausted T cells.1 Margetuximab, an investigational Fc-engineered anti-HER2 monoclonal antibody, has similar HER2 binding and antiproliferative properties to trastuzumab, but with enhanced Fc-mediated effector function. In vitro studies have demonstrated upregulation of LAG-3/PD-L1 expression on immune cells after margetuximab exposure, along with enhanced lytic activity of immune cells primed by margetuximab in the presence of tebotelimab. Methods This study characterizes safety, PK/PD, and preliminary antitumor activity of tebotelimab plus margetuximab in patients with advanced HER2+ malignancies. A one-step 3+3 dose escalation phase of tebotelimab (300 and 600 mg) combined with margetuximab 15 mg/kg, both every 3 weeks, was followed by cohort expansion of patients with breast, gastric or gastroesophageal, and other HER2+ tumors. Results At data-cutoff, 31 patients (2.0 median lines of prior therapy; 64.5% with prior HER2-directed therapy) were treated. Median duration of treatment is 10.3 weeks with 17 patients remaining on treatment. No maximum tolerated dose was defined. Treatment-related adverse events (TRAEs) occurred in 23/31 (74.2%) patients, most commonly diarrhea (n=6), nausea, ALT increased (n=5, each), AST increased, and myalgia (n=4, each). The rate of Grade 3 TRAEs was 19.4%, with no Grade 4–5 TRAEs observed. Immune-related AEs were consistent with events observed with anti-PD-1 antibodies and were manageable with supportive treatment. Among 20 response-evaluable patients (i.e., received on-treatment scan), 8 objective responses (6 confirmed) per RECIST v1.1 have been observed, including a confirmed complete response (cholangiocarcinoma) and 7 partial responses (breast [2], microsatellite stable colorectal cancer [2], esophageal adenocarcinoma [1], ovarian cancer [1], and microsatellite stable gastroesophageal junction carcinoma).1 Immunohistochemistry (IHC) of available baseline tumor specimens (n=17) demonstrated low PD-L1 expression with combined positive scores of either 0 (n=16) or 1 (n=1, colorectal cancer). Investigations into other potential correlative biomarkers, including LAG-3 and PD-1 by IHC and gene expression profiling by NanoString, remain ongoing. Conclusions Tebotelimab in combination with margetuximab has demonstrated an acceptable safety profile and encouraging early evidence of anti-tumor activity, with a preliminary overall response rate (ORR) of 40% (8/20) [including unconfirmed responses] among late-line patients with various advanced HER2+ malignancies. Trial Registration NCT03219268 Ethics Approval This study was approved by each Institution’s Ethics Board prior to enrollment of subjects. Reference Luke J, et al. A phase I, first-in-human, open-label, dose-escalation study of MGD013, a bispecific DART molecule binding PD–1 and LAG–3, in patients with unresectable or metastatic neoplasms. J Clin Oncol38: 2020 (suppl; abstr 3004).

Background Interleukin-10 (IL-10) is a multifunctional cytokine that can mediate immune suppression or activation depending on the immunological context. Mouse studies have demonstrated that blockade of IL-10 enhances immune response against tumors and chronic viral infections;1 2 intriguingly, high concentrations of long-acting, pegylated IL-10 have also shown anti-tumor activity.3 Here we investigated IL-10 and IL-10 receptor-alpha (IL-10RA) expression profiles in normal and tumor tissues as well as the immunological effects of modulating the IL-10 pathway via antibody-mediated blockade of IL-10RA. Methods IL-10 and IL-10RA mRNA are expressed by several tumors, including renal, lung, breast, and colon cancers. Fluorescent in-situ hybridization revealed that the majority of IL-10RA was expressed by CD3-negative tumor-infiltrating cells, localized in close proximity to T cells in the tumor microenvironment (TME). Immunohistochemistry studies confirmed expression of IL-10RA in the TME, while no expression was detected in healthy tissues. Furthermore, dissociated tumor cells produced biologically active levels of IL-10 in culture. Results Monoclonal antibodies (mAbs) against IL-10RA prevented IL-10 signaling and enhanced release of IL-12 by monocyte-derived dendritic cells activated with suboptimal LPS concentrations. The effect of IL-10RA blockade was greater than that observed with IL-10 neutralizing mAbs. In mixed lymphocyte reactions and superantigen-driven T-cell activation, IL-10RA blockade enhanced IL-2 secretion by T lymphocytes. Consistent with earlier observations in mouse models,4 the effect of IL-10RA blockade was nonredundant with blockade of the PD-1/PD-L1 axis, resulting in enhanced IL-2 and interferon-gamma secretion by T cells when both pathways were inhibited. Blockade of IL-10RA during CD3-redirected in vitro killing of tumor cells by PBMC induced IL-12 release as well as upregulation of CD86 and HLA-DR by CD3-negative cells. In in vitro dissociated tumor cells, IL-10RA blockade induced release of IL-2, interferon-gamma and other proinflammatory cytokines; additional PD-1/PD-L1 axis blockade further enhanced cytokine release. Conclusions In summary, antibody-mediated IL-10RA blockade can potentiate immune activation in the dissociated tumor cells and may be a valuable addition to cancer immunotherapies, including redirected T-cell killing and checkpoint blockade. References Vicari A, et al. J. Exp. Med 2002;196:541–549. Ejrnaes M, et al. J Exp Med 2006;203:2461–72. Emmerich J, et al. Cancer Res 2012. 72:3570–3581. Brooks D, et al. Proc Natl Acad Sci U S A 2008;105:20428–33.

Carboniferous (Mississippian) limestones of the Burren are cross-cut by sub-vertical veins, from 1µm up to 50cm thick, defining a strongly clustered and scale-indepen-dent system in which predominantly N-S veins are transected by longer NNE-trending veins. Vein infills mainly comprise of calcite, but with subordinate amounts of quartz, sulphide (mainly galena and sphalerite) and fluorite also occurring, particularly in the south-central part of the area. Thinner and shorter veins are planar and discontinuous in plan view, sometimes forming en-echelon arrays, with thicker veins forming better connected and more complex structures which extend for several kilometres across the Burren region. Veins with ‘exotic’ infills are generally both longer and thicker, and they appear to be spatially associated with, or up to 5km to the north of, a 5km wide zone of ENE-trending Variscan monoclinal folding. Individual veins are vertically persistent, and the same structures are seen throughout the exposed ca 1200m thick Carboniferous sequence, from Tournaisian limestones through to Serpukhovian-Bashkirian siliciclastics. The veins are mainly extensional, sometimes with a component of sinistral displacement particularly on NNE-trending veins, displaying fibrous growth through to hydraulic fracturing and brecciation. Their formation is attributed to the valving of overpressured fluids within Mississippian basins during N-S Variscan compression. Pb isotope analysis supports a model in which sulphide infills are scavenged from underlying basement rocks or hydrothermal Zn-Pb mineralisation during the tectonic inversion of post-rift sequences overlying Lower Carboniferous normal faults. Environmental Protection Agency European Commission - European Regional Development Fund Science Foundation Ireland Geological Survey Ireland/DCCAE Postdoctoral Fellowship Programme

Introduction: Notwithstanding recent progress, acute myeloid leukemia (AML) remains an incurable disease, particularly in patients (pts) with relapsing/refractory disorder or ineligible for intensive induction therapy (unfit pts). Redirected T-cell mediated killing via CD3-engaging bispecific molecules may offer an alternate therapeutic opportunity for aggressive disease or unfit pts. Flotetuzumab, a continuous infusion CD123 x CD3 DART molecule, has shown preliminary single-agent activity in pts refractory to induction therapy (Uy et al., Blood. 2021, 137:751). CD123, the IL-3 receptor alpha chain, is expressed by both leukemic blasts and leukemic stem cells and is a suitable therapeutic target in AML (Testa et al., Cancers (Basel). 2019, 11:1358). MGD024 is an Fc-bearing CD123 x CD3 DART molecule designed for prolonged circulating half-life and intermittent delivery. MGD024 was also designed with a CD3-binding arm with reduced affinity to diminish the propensity for cytokine release compared to flotetuzumab. The potentially improved tolerability and dosing convenience of MGD024 may provide a framework for introducing T-cell immunotherapy in early-stage AML or unfit pts. To explore whether MGD024 could complement AML standard of care (SOC), we investigated combination therapy in mouse models. Materials and Methods: The DART molecules, flotetuzumab and MGD024, shared identical CD123 (humanized 7G3) and CD3 (humanized XR32) Fv arms, save for a mutation in the anti-CD3 arm of MGD024 that decreases its affinity for the CD3-epsilon chain. While flotetuzumab has no Fc domain, MGD024 includes an ala-ala-mutated human IgG1 Fc that extends its circulating half-life via the neonatal Fc receptor-mediated salvage pathway together with impairing binding to Fc-gamma receptors and complement. An IgG1-ala-ala Fc-bearing version of flotetuzumab (RES234M1.1) was also engineered to allow delivery at identical time intervals as MGD024 and avoid continuous infusion in experimental animals. MHC class I-null, NOD/SCID/IL2R-gamma-null mice were reconstituted with human PBMC (8x10 6 cells/mouse, retro orbital). Two human AML cell lines expressing low or high levels of CD123 (KG1a < MOLM-13) were implanted SC at 2.5 x 10 6 (KG1a) or 5 x 10 6 (MOLM-13) cells/mouse. Treatments (IV, IP or PO by gavage, as indicated) were initiated when tumor volumes reached ~150 mm 3, with volumes recorded weekly or twice weekly thereafter. Results and Conclusions: Consistent with its decreased affinity for CD3, MGD024 demonstrated reduced in vitro potency in killing CD123-positive target cells compared to flotetuzumab or RES234M1.1, but proportionally greater reduction in cytokine release. MGD024, however, achieved maximal cytolytic activity as flotetuzumab or RES234M1.1, albeit at increased concentrations. Similarly, MGD024 showed reduced potency in vivo against CD123-positive tumors compared to RES234M1.1; nevertheless, tumor growth reduction of the same magnitude as that observed with RES234M1.1 was attained at higher doses of MGD024 (0.5-1 mg/kg IV 2QW MGD024 vs. 0.05-0.1 mg/kg IV 2QW RES234M1.1, depending on the model). Reduced cytokine release was also observed with MGD024 compared to RES234M1.1 in vivo. To explore MGD024 suitability for combination therapy, sub-active doses of cytarabine (CYT, 10 mg/kg IV 2QW or 7.5-10 mg/kg IP QD), venetoclax (VEN, range 10-80 mg/kg PO QD), or azacitidine (AZA, 2 mg/kg PO QD) were co-administered with suboptimal regimens of MGD024 (range 0.005-0.1 mg/kg IV 2QW, depending on the model). Complete or near complete tumor elimination was observed with the combination of suboptimal MGD024 and CYT or VEN. In contrast, AZA, at the dose tested, did not contribute to the antitumor effect of MGD024. CYT, VEN or AZA did not inhibit a fully active dose of MGD024, confirming no detrimental impact of the SOC agents at the doses employed on the effector cell population engaged by the DART molecule. All treatments were well tolerated, as indicated by body weigh profiles across treatment groups. These data support clinical exploration of the combination of MGD024 with SOC in patients with AML. An investigational new drug (IND) application of MGD024 in pts with selected relapsed or refractory hematologic malignancies is planned. Disclosures Alderson: MacroGenics: Current Employment, Current equity holder in publicly-traded company, Current holder of individual stocks in a privately-held company, Current holder of stock options in a privately-held company. Huang: MacroGenics: Current equity holder in publicly-traded company, Current holder of individual stocks in a privately-held company, Current holder of stock options in a privately-held company, Ended employment in the past 24 months. Zhang: MacroGenics: Current Employment, Current equity holder in publicly-traded company, Current holder of individual stocks in a privately-held company, Current holder of stock options in a privately-held company. Li: MacroGenics: Current Employment, Current equity holder in publicly-traded company, Current holder of individual stocks in a privately-held company, Current holder of stock options in a privately-held company. Kaufman: MacroGenics: Current Employment, Current equity holder in publicly-traded company, Current holder of individual stocks in a privately-held company, Current holder of stock options in a privately-held company. Diedrich: MacroGenics: Current Employment, Current equity holder in publicly-traded company, Current holder of individual stocks in a privately-held company, Current holder of stock options in a privately-held company. Moore: MacroGenics: Current Employment, Current equity holder in publicly-traded company, Current holder of individual stocks in a privately-held company, Current holder of stock options in a privately-held company. Bonvini: MacroGenics: Current Employment, Current equity holder in publicly-traded company, Current holder of individual stocks in a privately-held company, Current holder of stock options in a privately-held company.

Background: MGD013 is an investigational, first-in-class, Fc-bearing bispecific tetravalent DART molecule designed to bind PD-1 and LAG-3 and sustain/restore the function of exhausted T cells (1). MGD013 demonstrates in vitro ligand blocking properties and improved T-cell responses beyond that observed with anti-PD-1 and anti-LAG-3 benchmark antibodies alone or in combination. PD-1 targeted therapy with nivolumab in patients (pts) with relapsed or refractory (R/R) diffuse large B cell lymphoma (DLBCL) has yielded modest efficacy (2). LAG-3, highly expressed in DLBCL (3), has emerged as another therapeutic target of interest in this population with continued unmet need. Methods: This study characterizes the safety, tolerability, dose-limiting toxicities, maximum tolerated dose, PK/PD, and antitumor activity of MGD013 in pts with advanced solid and hematologic malignancies. R/R DLBCL pts are being treated at the recommended Phase 2 dose of 600 mg every two weeks in Cohort Expansion. Results: At data-cutoff, 17 DLBCL pts received MGD013 (2.5 median prior lines of therapy, 41.2% with prior CAR-T therapy). Treatment-related adverse events (TRAEs) occurred in 11/17 (64.7%) pts, with no same TRAE occurring in > 1 patient except pyrexia (n=3). One grade ≥ 3 TRAE occurred (pneumonia), and no events of tumor lysis syndrome were observed. Among 11 response-evaluable pts (i.e. received at least one on-treatment scan), 1 complete response (CR) and 3 partial responses (PRs) per the Lugano Classification were observed. Analyses of available pre-treatment tumor biopsy samples corresponding to responding pts demonstrated relatively high levels of LAG-3, PD-1, and PD-L1 by IHC. More comprehensive translational analyses were undertaken for the pt with CR, observed after a single MGD013 infusion in a 28-year-old male in relapse 6 months after CD19-directed CAR T-cell therapy. In contrast to a pre-CAR T biopsy specimen demonstrating no LAG-3 or PD-1 expression, IHC analysis of a lymph node specimen biopsied post-CAR T and prior to MGD013 treatment revealed high levels of both LAG-3 and PD-1 on both infiltrating T-cells and malignant B-cells. Consistent with CD19 CAR T resistance, no CD19 expression was evident. MHC class II and PD-L1 expression were observed, which when bound to LAG-3 and PD-1, respectively, form immune checkpoints that can decrease T cell function. Following MGD013 treatment, serum IFN-γ markedly increased to >140-fold above baseline. No significant changes were observed for IL-6 or IL-2. Expansion of circulating CD3+CD8+ and CD3+CD4-CD8- T-cell subsets and associated cytolytic markers (i.e., perforin, granzyme B) were observed following MGD013 treatment. The patient underwent allogeneic stem cell transplant (allo-SCT) and remains in remission 14 months post-MGD013 and 12 months post-allo-SCT. Further correlative biomarker analyses are underway in the ongoing clinical trial. Conclusion: MGD013, a novel molecule designed to coordinately block PD-1 and LAG-3, has preliminarily demonstrated an acceptable safety profile and encouraging early evidence of anti-tumor activity in R/R DLBCL pts with and without prior treatment with CAR T. Biomarker analyses confirmed expression of both PD-1 and LAG-3 axes in responding pts with evidence of pharmacodynamic responses consistent with the ability of MGD013 to enhance T-cell function. References: 1. Luke JJ, Patel MR, Hamilton E, et al. A phase I, first-in-human, open-label, dose-escalation study of MGD013, a bispecific DART molecule binding PD-1 and LAG-3, in patients with unresectable or metastatic neoplasms. J Clin Oncol 38: 2020 (suppl; abstr 3004). 2. Keane C, Law SC, Gould C, et al. LAG3: a novel immune checkpoint expressed by multiple lymphocyte subsets in diffuse large B-cell lymphoma. Blood Adv. 2020;4(7):1367-1377. doi:10.1182/bloodadvances.2019001390 3. Ansell SM, Minnema MC, Johnson P, et al., Nivolumab for Relapsed/Refractory Diffuse Large B-Cell Lymphoma in Patients Ineligible for or Having Failed Autologous Transplantation: A Single-Arm, Phase II Study. J Clin Oncol, 2019. 37(6): p. 481-489. Disclosures Wang: Verastem Oncology: Membership on an entity's Board of Directors or advisory committees; Curio Science: Honoraria; Putnam LLC: Honoraria. Asch:Astellas Pharma: Research Funding; Cartesian: Research Funding; Forty Seven: Research Funding; Juno: Research Funding; MacroGenics: Research Funding; MEI Pharma: Patents & Royalties: Provisional patent submitted (I), Research Funding. Hamad:Novartis: Honoraria; Abbvie: Honoraria. Weickhardt:Merck: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; BMS: Membership on an entity's Board of Directors or advisory committees, Research Funding; Ipsen: Membership on an entity's Board of Directors or advisory committees; Pfizer: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding. Ulahannan:Merck Co. Inc: Research Funding; Bayer: Membership on an entity's Board of Directors or advisory committees; G1 Therapeutics, Inc.: Research Funding; ArQule, Inc.: Research Funding; Evelo Biosciences, Inc.: Research Funding; Incyte: Membership on an entity's Board of Directors or advisory committees, Research Funding; Klus Pharma, Inc.: Research Funding; Isofol: Research Funding; GlaxoSmithKline GSK: Research Funding; Mersana Therapeutics: Research Funding; Macrogenics: Research Funding; Celgene Corporation: Research Funding; Boehringer Ingelheim: Research Funding; Array: Membership on an entity's Board of Directors or advisory committees; Ciclomed LLC: Research Funding; AbbVie, Inc.: Research Funding; AstraZeneca: Research Funding; Bristol-Myers Squibb: Research Funding; Syros: Membership on an entity's Board of Directors or advisory committees; Exelxis: Membership on an entity's Board of Directors or advisory committees. Koucheki:MacroGenics, Inc.: Current Employment, Current equity holder in publicly-traded company. Sun:MacroGenics, Inc.: Current Employment, Current equity holder in publicly-traded company. Li:MacroGenics, Inc.: Current Employment, Current equity holder in publicly-traded company. Chen:MacroGenics, Inc.: Current Employment, Current equity holder in publicly-traded company. Zhang:MacroGenics, Inc.: Current Employment, Current equity holder in publicly-traded company. Muth:MacroGenics, Inc.: Current Employment, Current equity holder in publicly-traded company. Kaminker:MacroGenics, Inc.: Current Employment, Current equity holder in publicly-traded company. Moore:MacroGenics, Inc.: Current Employment, Current equity holder in publicly-traded company. Sumrow:MacroGenics, Inc.: Current Employment, Current equity holder in publicly-traded company.

Although improvements in genetic analysis have greatly enhanced our understanding of the mechanisms behind pancreatitis, it continues to afflict many families for whom the hereditary factors remain unknown. Recent evaluation of a patient with a strong family history of pancreatitis prompted us to reexamine a large kindred originally reported over 50 years ago with an autosomal-dominant inheritance pattern of chronic pancreatitis, diabetes, and pancreatic adenocarcinoma. Whole-exome sequencing analysis identified a rare missense mutation in the gene encoding pancreas-specific protease elastase 3B (CELA3B) that cosegregates with disease. Studies of the mutant protein in vitro, in cell lines, and in CRISPR-Cas9–engineered mice indicate that this mutation causes translational upregulation of CELA3B, which, upon secretion and activation by trypsin, leads to uncontrolled proteolysis and recurrent pancreatitis. Although lesions in several other pancreatic proteases have been previously linked to hereditary pancreatitis, to our knowledge, this is the first known instance of a mutation in CELA3B and a defect in translational control contributing to this disease.

The overuse and misuse of antibiotics affect patients in many ways, including by driving antibiotic resistance, a serious public health threat in the United States and around the world. To improve patient safety and address rising rates of resistance, an increasing number of health care facilities have created antibiotic stewardship programs (ASPs). ASPs have been successful in slowing the emergence of resistance and improving patient outcomes. However, there are serious geographic and resource barriers to ASP adoption in small community hospitals and critical access hospitals. Fortunately, many barriers can be overcome by using collaborative models to bring together key stakeholders, including large hospitals and health systems and academic medical centers; hospital associations; federal, state, and local public health organizations; and federal and state offices of rural health. These stakeholders are ideally positioned to assist with stewardship efforts in small community and critical access hospitals and, in doing so, can improve patient safety while stemming the spread of resistant bacteria.

Background: Margetuximab (M) has demonstrated anti-tumor activity in patients with advanced HER2+ gastric cancer and PFS superiority to trastuzumab (T) in pre-treated metastatic HER2+ breast cancer patients (1). Similar to T, M inhibits HER2 signaling; additionally, M enhances engagement with the activating Fcγ receptor (FcR)IIIA, while diminishing interaction with the inhibitory FcRIIB. Given the role of FcRs in coupling innate and adaptive immune responses, clinical studies of M in combination with PD-1 or PD-1 plus LAG-3 blockade have been initiated (2,3). The combination of M with tebotelimab, an investigational bispecific DART® molecule blocking PD-1 and LAG-3, has demonstrated encouraging early evidence of anti-tumor activity in patients with advanced HER2+ malignancies, including those with PD-L1-negative tumors (3). Here we present data supporting the potential mechanisms underlying the anti-tumor activity elicited by M and tebotelimab combination therapy. Methods: PBMCs were challenged with HER2-expressing tumor cell lines in the presence of M or T. Immune modulation was assessed by analysis of immune cell phenotype, gene expression profiles, and cytokine secretome. Functional activity of immune cells following exposure to the combination of M with tebotelimab was evaluated. Results: M induced greater upregulation relative to T of both co-stimulatory (eg 4-1BB) and/or checkpoint molecules (eg PD-L1 and LAG-3) on NK cells and/or monocytes in PBMCs cocultured with HER2-expressing tumor cells. Compared with T, M also induced greater proinflammatory cytokine release, including IFNγ that in turn upregulates PD-L1 and MHC class I expression on co-cultured tumor cells. These effects were not observed with an Fc-null version of M or were inhibited by the addition of FcR-blocking Abs, demonstrating Fc:FcR dependency of M-induced immune activation. Interestingly, CD8 T cells, which lack FcR, also exhibited LAG-3 upregulation in the above cultures, suggesting the effects of M can also be mediated via a cascade mechanism. An IFNγ neutralizing mAb reduced tumor cell-associated PD-L1 and MHC class I upregulation, but not LAG-3 induction on T-cells. PBMCs, pre-treated with a combination of M and tebotelimab in the presence of HER2-expressing tumor cells, demonstrated enhanced cytolytic activity against both the canonical NK cell target, K562, or M-opsonized HER2-positive tumor cells. Conclusions: M, through an Fc-dependent mechanism, can upregulate checkpoint molecules on NK-cells, CD8 T cells and tumor cells, thereby sensitizing them to immune checkpoint blockade. In turn, PD-1 and LAG-3 blockade by tebotelimab enhances M-mediated NK cell cytolytic activity in vitro. References: 1. Rugo et al., 2019 ASCO oral presentation 2. Catenacci et al., 2020 Lancet Oncology 21(8):1066-76 3. Luke et al., 2020 ASCO oral presentation Citation Format: Xiaoyu Zhang, Haiquan Li, Yinhua Yang, Qihong Xu, Tom Kaufman, Doug Smith, Jeffrey Nordstrom, Ezio Bonvini, Paul Moore. Enhanced HER2-dependent immune activation by margetuximab, an investigational Fc-engineered anti-HER2 mAb, supports combination with checkpoint blockade [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1555.

Master regulators of the unfolded protein response (UPR)—IRE1α and PERK— promote adaptation or apoptosis depending on levels of endoplasmic reticulum (ER) stress. While the UPR is activated in many cancers, its effects on tumor growth remain unclear. Derived from endocrine cells, pancreatic neuroendocrine tumors (PanNETs) universally hypersecrete one or more peptide hormones, likely sensitizing these cells to high ER protein-folding stress. For the nearly 1,500 Americans diagnosed with PanNETs annually, surgery is the only potentially curative treatment; however the five-year survival is extremely low for those who develop metastatic disease. To assess whether targeting the UPR is a viable therapeutic strategy, we analyzed human PanNET samples and found evidence of elevated ER stress and UPR activation. We then used genetic and pharmacologic approaches to modulate IRE1α and PERK in cultured cells and xenograft and spontaneous genetic (RIP-Tag2) mouse models of PanNETs. We found that UPR signaling is optimized for adaptation and that inhibiting either IRE1α or PERK leads to hyperactivation and apoptotic signaling through the reciprocal arm, thereby halting tumor growth and survival. Our results provide a strong rationale for therapeutically targeting the UPR in PanNETs and other cancers experiencing elevated ER stress.SignificanceThe unfolded protein response (UPR) is upregulated in human pancreatic neuroendocrine tumors and its genetic or pharmacological inhibition significantly reduces tumor growth in preclinical models, providing strong rationale for targeting the UPR in neoplasms with elevated ER stress.

Under misfolded protein stress, the endoplasmic reticulum (ER) activates the unfolded protein response (UPR) to restore homeostasis, or commits the cell to apoptosis. A study now uncovers how the UPR is governed by the circadian clock to adjust ER protein-folding capacity to metabolic demand and protect against liver damage.

The School of Creative Arts & Technologies at Ulster University (Magee) has brought together the subject of computing with creative technologies, cinematic arts (film), drama, dance, music and design in terms of research and education. We propose here the development of a flagship computer software platform, SceneMaker, acting as a digital laboratory workbench for integrating and experimenting with the computer processing of new theories and methods in these multidisciplinary fields. We discuss the architecture of SceneMaker and relevant technologies for processing within its component modules. SceneMaker will enable the automated production of multimodal animated scenes from film and drama scripts or screenplays. SceneMaker will highlight affective or emotional content in digital storytelling with particular focus on character body posture, facial expressions, speech, non-speech audio, scene composition, timing, lighting, music and cinematography. Applications of SceneMaker include automated simulation of productions and education and training of actors, screenwriters and directors. © ICST Institute for Computer Sciences, Social Informatics and Telecommunications Engineering 2017.

Background The purpose of this study is to determine if antioxidant supplementation influences the incidence of atrial arrhythmias in trauma intensive care unit (ICU) patients. Materials and methods In this retrospective pre-post study, critically ill injured patients aged ≥18 years, admitted to a single-center trauma ICU for ≥48 hours were eligible for inclusion. The control group consists of patients admitted from January 2000 to September 2005, before routine antioxidant supplementation in our ICU. The antioxidant group consists of patients admitted from October 2005 to June 2011 who received an antioxidant protocol for ≥48 hours. The primary outcome is the incidence of atrial arrhythmias in the first 2 weeks of hospitalization or before discharge. Results Of the 4699 patients, 1622 patients were in the antioxidant group and 2414 patients were in the control group. Adjusted for age, sex, year, injury severity, past medical history, and medication administration, the unadjusted incidence of atrial arrhythmias was 3.02% in the antioxidant group versus 3.31% in the control group, with no adjusted difference in atrial arrhythmias among those exposed to antioxidants (odds ratio: 1.31 [95% confidence interval: 0.46, 3.75], P = 0.62). Although there was no change in overall mortality, the expected adjusted survival of patients in those without antioxidant therapy was lower (odds ratio: 0.65 [95% confidence interval: 0.43, 0.97], P = 0.04). Conclusions ICU antioxidant supplementation did not decrease the incidence of atrial arrhythmias, nor alter the time from admission to development of arrhythmia. A longer expected survival time was observed in the antioxidant group compared with the control group but without a change in overall mortality between groups.

OBJECTIVE: To determine if daily respiratory status improved more in extremely low gestational age premature infants after diuretic exposure compared with those not exposed in modern neonatal intensive care units (NICU). STUDY DESIGN: The Prematurity and Respiratory Outcomes Program (PROP) was a multi-center observational cohort study of 835 extremely premature infants, gestational ages 23 0/7–28 6/7 weeks, enrolled in the first week of life from 13 U.S. tertiary NICUs. We analyzed the PROP study daily medication and respiratory support records of infants up to 34 weeks postmenstrual age. We determined whether there was a temporal association between administration of diuretics and an acute change in respiratory status in premature infants in the NICU, using an ordered categorical ranking of respiratory status. RESULTS: Infants in the diuretic exposed group of PROP were of lower mean gestational age and lower mean birth weight (p 1) for each day after the initial day of diuretic exposure. CONCLUSIONS: Our analysis did not support the ability of diuretics to substantially improve the extremely premature infant’s respiratory status. Further study of both safety and efficacy of diuretics in this setting are warranted. TRIAL REGISTRATION: Clinicaltrials.gov NCT01435187

Dysregulation of ADAM9, a member of the ADAM (a disintegrin and metalloproteinase) family of proteases, has been implicated in tumor progression and metastasis, as well as pathological neovascularization. ADAM9 overexpression correlates with poor prognosis in multiple cancers. We have shown that ADAM9 is overexpressed in multiple solid tumor indications and that anti-ADAM9 antibodies are efficiently internalized and degraded by tumor cell lines making ADAM9 an attractive target for antibody-drug conjugate (ADC) development. Here, we describe IMGC936, the first ADAM9-targeting ADC to enter preclinical development. IMGC936 is comprised of a high-affinity humanized antibody site-specifically conjugated to DM21, a next-generation linker-payload that combines a maytansinoid microtubule-disrupting payload with a stable peptide linker at a drug-antibody ratio of two. To maximize the potential for IMGC936 activity, the M252Y/S254T/T256E (YTE) mutation was introduced into the CH2 domain of the antibody to increase in vivo plasma half-life and exposure. In vitro studies demonstrated targeted cytotoxicity of IMGC936 across a panel of ADAM9-positve tumor cell lines with activity at least 2 logs greater than a non-targeting conjugate. Consistent with the in vitroactivity, an anti-ADAM9-DM21 conjugate displayed compelling anti-tumor activity in multiple xenograft models representing non-small cell lung, gastric and colorectal cancers. For example, in the EBC-1 non-small cell lung cancer subcutaneous xenograft model with only moderate ADAM9 expression (H-score of 130), anti-ADAM9-DM21 not only induced tumor growth delay but produced complete and durable remissions in 6/6 mice following a single intravenous dose of 8.6 mg Ab/kg (100 ug DM21/kg). IMGC936 demonstrated a favorable pharmacokinetic profile with good conjugate stability in non-human primates. Importantly, IMGC936 was well-tolerated following repeat dosing in cynomolgus monkeys with no ADAM9 target-related toxicities identified at doses exceeding the levels required for anti-tumor activity in murine xenograft models. Based on the totality of the preclinical data, IMGC936 represents a promising therapeutic candidate to target a wide range of ADAM9-expressing tumors. Citation Format: Stuart Hicks, Deryk Loo, Kerstin Sinkevicius, Juniper Scribner, Bhaswati Barat, Nicholas Yoder, Christopher Espelin, Marian Themeles, Francine Chen, Jacquelynn Lucas, Jennifer Brown, Bahar Matin, Megan Fuller, Jenny Lee, Paulin Salomon, Juliet Costoplus, Sadiqa Yancey, Gundo Diedrich, Sergey Gorlatov, Thomas Son, Michael Chiechi, Pam Li, Michael Spliedt, Valentina Ciccarone, Jeff Hooley, Nadia Gantt, James Tamura, Kerry Donahue, Paul Moore, Syd Johnson, Thomas Chittenden, Richard Gregory, Ezio Bonvini. IMGC936, a first-in-class ADAM9-targeting antibody-drug conjugate, demonstrates promising anti-tumor activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1533.

Margetuximab (M) is an investigational Fc-engineered anti-HER2 monoclonal antibody (mAb) with potential for greater immune-mediated anti-tumor activity than trastuzumab (T). M and T bind the same subdomain IV epitope, but M binds with higher affinity to activating Fc receptor, CD16A (FcγRIIIA), and lower affinity to inhibitory Fc receptor, CD32B (FcγRIIB). Pertuzumab (P) binds to a subdomain II epitope and has the same wild type Fc domain as T. We compared in vitro properties of M, T and P and combinations of M+P and T+P to evaluate Fc domain contributions to anti-HER2 mAb mediated activities. By surface plasmon resonance, M, T and P bound to the HER2 extracellular domain with similar high affinity. Binding affinities of M and T were unchanged if P was pre-bound, indicating lack of interaction. M, T and P exhibited comparable binding to HER2-expressing cell lines. Anti-proliferative activities of M and T toward HER23+ cells (N87, SKBR3 or BT474) in the absence or presence of ligands (EGF or HRG1β) were similar, whereas P exhibited weaker activity. In the absence of ligands, M+P or T+P were comparably as active as M or T alone. In presence of ligands, M+P and T+P were generally more active than M or T alone. Antibody-dependent cell-mediated cytotoxicity (ADCC) was evaluated with N87 and SKBR3 target cells (HER23+, expressing a reporter gene to measure viability) and NK effector cells from donors with differing CD16A158 genotypes. M was 7- to 84-fold more potent than T, as well as 69- to 744-fold more potent than P. Greater differences generally were seen with effector NK cells bearing the more common CD16A158 FF and VF genotypes than with the less common, high affinity CD16A158 VV genotype. While addition of P enhanced mean potency of T by 2- to 3-fold, no such effect was seen with M; however, M or M+P was 7- to 25-fold more potent than T+P. Results with JIMT-1 cells (HER22+) were similar, albeit with lower differential than with HER23+ target cells. NK cells were monitored by flow cytometry following incubation of peripheral blood mononuclear cells with HER2+ target cells and mAbs. M induced greater expression of markers of activation (CD137), cytolytic capability (granzyme B, perforin) and proliferation (Ki67) than T. An anti-HER2 mAb with an inactivated Fc domain was ineffective, indicating that induced changes in NK cells were Fc-dependent. In conclusion, M and T bind HER2 with similar affinity and exhibit comparable anti-proliferative activity that is enhanced by addition of P if ligands (EGF or HRG1β) are present. Relative to T, M mediates superior ADCC with effector cells of all CD16A genotypes and promotes greater NK cell activation and expansion. The M+P combination maintains superior ADCC compared to the T+P combination. Citation Format: Liqin Liu, Yinhua Yang, Robert Burns, Jonathan Li, Haiquan Li, Sergey Gorlatov, Paul Moore, Jeffrey Nordstrom. Margetuximab mediates greater Fc-dependent anti-tumor activities than trastuzumab or pertuzumab in vitro [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1538.

Introduction: Trophoblast glycoprotein 5T4 is expressed on the cell surface of multiple cancers yet sparingly on normal adult tissues. CD137 (4-1BB) is a co-stimulatory molecule expressed by activated T and NK cells that support cell activation, proliferation and survival. Previously we described bispecific tumor associated antigen x CD137 DART® molecules limiting CD137 mediated immunostimulation to the tumor microenvironment (AACR2017) as an approach to reduce the unwanted systemic CD137 effects associated with agonistic anti-CD137 mAbs. We subsequently identified a bispecific TRIDENT format bearing bivalent CD137 and monovalent tumor antigen engagement that provides maximal CD137 activation in a tumor antigen anchored manner. Here we apply that format to generate a 5T4 x CD137 x CD137 TRIDENT molecule that promotes CD137 activation in proximity to 5T4-expressing tumor cells leading to enhanced T-cell co-stimulation, proliferation and tumor cell killing activity when combined with CD3-engaging tumor-targeting bispecific DART molecules. Methods: TRIDENT molecules were constructed comprising bivalent CD137 binding and monovalent 5T4 binding. Binding properties were evaluated by surface plasmon resonance (SPR) and flow cytometry. Signaling was assessed using a NF-κB luciferase reporter cell line expressing CD137. Co-stimulatory activity was characterized with primary human or cynomolgus monkey T cells. T cells were incubated with or without antigen-expressing cells and submaximal concentrations of either anti-CD3 bead or CD3-engaging tumor-targeting bispecific DART molecules. Results: SPR and flow cytometry analyses demonstrate that 5T4 x CD137 x CD137 TRIDENT molecule binds human and monkey target antigens. The 5T4 x CD137 x CD137 TRIDENT molecule induces CD137 signaling in a reporter cell line and promotes cytokine release in primary human & cynomolgus monkey T-cell assays in the presence of 5T4+ tumor cells. In the absence of 5T4+ tumor cells, the TRIDENT molecule lacks agonistic activity. Furthermore, 5T4 x CD137 x CD137 TRIDENT molecule enhances T-cell proliferation and tumor cell cytolysis induced by CD3-targeted DART molecules. Consistent with preferential induction of CD137 by CD8 T-cells, 5T4 x CD137 x CD137 TRIDENT increases the fraction of CD8+ central memory and effector memory T cells in the presence 5T4+ tumor cells. Conclusions: 5T4 x CD137 x CD137 TRIDENT molecule promotes T-cell co-stimulation in a tumor antigen-dependent manner offering an opportunity to target CD137 immunostimulation, while potentially limiting non-specific systemic T-cell activation and related side effects. Citation Format: Liqin Liu, Ling Huang, Vatana Long, Yinhua Yang, Robert Burns, Jonathan Li, Jennifer DiChiara, Qihong Xu, Arin Whiddon, Chia-Ying K Lam, Jim Tamura, Valentina Ciccarone, Syd Johnson, Ezio Bonvini, Paul Moore. Tumor-antigen 5T4-dependent activation of the CD137 costimulatory pathway by bispecific 5T4 x CD137 x CD137 TRIDENT™ molecules [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 554.