KLRG1 Cell Depletion as a Novel Therapeutic Strategy in Patients with Mature T-Cell Lymphoma Subtypes.

PURPOSE: Develop a novel therapeutic strategy for patients with subtypes of mature T-cell and NK-cell neoplasms. EXPERIMENTAL DESIGN: Primary specimens, cell lines, patient-derived xenograft models, commercially available, and proprietary anti-KLRG1 antibodies were used for screening, target, and functional validation. RESULTS: Here we demonstrate that surface KLRG1 is highly expressed on tumor cells in subsets of patients with extranodal NK/T-cell lymphoma (ENKTCL), T-prolymphocytic leukemia (T-PLL), and gamma/delta T-cell lymphoma (G/D TCL). The majority of the CD8+/CD57+ or CD3-/CD56+ leukemic cells derived from patients with T- and NK-large granular lymphocytic leukemia (T-LGLL and NK-LGLL), respectively, expressed surface KLRG1. The humanized afucosylated anti-KLRG1 monoclonal antibody (mAb208) optimized for mouse in vivo use depleted KLRG1+ TCL cells by mechanisms of ADCC, ADCP, and CDC rather than apoptosis. mAb208 induced ADCC and ADCP of T-LGLL patient-derived CD8+/CD57+ cells ex vivo. mAb208 effected ADCC of subsets of healthy donor-derived KLRG1+ NK, CD4+, CD8+ Tem, and TemRA cells while sparing KLRG1- naïve and CD8+ Tcm cells. Treatment of cell line and TCL patient-derived xenografts with mAb208 or anti-CD47 mAb alone and in combination with the PI3K-δ/γ inhibitor duvelisib extended survival. The depletion of macrophages in vivo antagonized mAb208 efficacy. CONCLUSIONS: Our findings suggest the potential benefit of a broader treatment strategy combining therapeutic antibodies with PI3Ki for the treatment of patients with mature T-cell and NK-cell neoplasms. See related commentary by Varma and Diefenbach, p. 2300.

Opportunities to improve population health: possibilities for healthier food environments.

The recent Covid-19 pandemic highlighted stark social inequalities, notably around access to food, nutrition and to green or blue space (i.e. outdoor spaces with vegetation and water). Consequently, obesity is socio-economically patterned by this inequality; and while the environmental drivers of obesity are widely acknowledged, there is currently little upstream intervention. We know that living with obesity contributes to increasing health inequalities, and places healthcare systems under huge strain. Our environment could broadly be described obesogenic, in the sense of supporting unhealthful eating patterns and sedentary behaviour. Evidence points to the existence of nearly 700 UK obesity policies, all of which have had little success. Obesity prevention and treatment has focused on educational and behavioural interventions targeted at individual consumers. A more sustainable approach would be to try and change the environments that promote less healthy eating and high energy intake as well as sedentary behaviour. Approaches which modify the environment have the potential to assist in the prevention of this complex condition. This review paper focuses on the role of wider food environments or foodscapes. While there is an imperfect evidence base relating to the role of the foodscape in terms of the obesity crisis, policy, practice, civic society and industry must work together and take action now, in areas where current evidence suggests change is required. Despite the current cost-of-living crisis, shaping the foodscape to better support healthful eating decisions has the potential to be a key aspect of a successful obesity prevention intervention.

Integrating the norm activation model and theory of planned behaviour to investigate farmer pro-environmental behavioural intention.

Sustainable agriculture requires cooperative and coordinated action across multiple sectors and policy domains. However, farmer-stakeholder behaviors and action remain pivotal to sustainable food system management in many rural development contexts. We assess farmer pro-environmental behavioral intention through the development and application of a novel integrated approach combining two dominant psychological theories of behavior change: the Norm Activation Model (NAM) and Theory of Planned Behavior (TPB). We apply this framework to targeted research with potato growers of Kerman Province in southeastern Iran, using survey data (sample n = 381) analyzed through structural equation modeling (SEM). The integrated NAM-TPB model provides insight into both pro-social and self-interested motivations for farmer pro-environmental behavioral intention, with the model explaining 77% of total variance. We found that three variables, Awareness of Consequence (AC), Perceived Behavioral Control (PBC), and Subjective Norms (SN) were the strongest indicators of pro-environmental behavioral intention. We recommend therefore that agricultural extension and state-led farmer education should first emphasize awareness-raising of negative environmental impacts of current farming practices within training programs, and second, improve social learning amongst farmer communities through sustained farmer community engagement, thus "stabilizing" a social norm of environmental protection amongst peer networks of agricultural workers.

Catalysis-Induced Phase Separation and Autoregulation of Enzymatic Activity.

We present a thermodynamically consistent model describing the dynamics of a multicomponent mixture where one enzyme component catalyzes a reaction between other components. We find that the catalytic activity alone can induce phase separation for sufficiently active systems and large enzymes, without any equilibrium interactions between components. In the limit of fast reaction rates, binodal lines can be calculated using a mapping to an effective free energy. We also explain how this catalysis-induced phase separation can act to autoregulate the enzymatic activity, which points at the biological relevance of this phenomenon.

Climate change and non-migration - exploring the role of place relations in rural and coastal Bangladesh.

Of growing research and policy interest are the experiences of people living under conditions of climate change-induced environmental stress, which either are unable to migrate (sometimes described as a 'trapped population') or are seemingly unwilling to do so (sometimes described as the 'voluntarily immobile'). This paper problematises and expands upon these binary categories: examining the complex dimensionality of non-migration as a form of place relations, explored through qualitative study of rural and coastal Bangladeshi communities. Through 60 semi-structured interviews of individuals from four communities in the Kalapara region, the analysis proffers four qualitatively derived and inter-related dimensions of voluntary and involuntary non-migration framed as a form of place relations. These four dimensions concern the following: (1) livelihood opportunities, (2) place obduracy, (3) risk perceptions, and (4) social-structural constraints, with the interplay between these elements explaining diverse non-migratory experiences. In our analysis, 'place obduracy' is introduced as a concept to describe the differential speed of environmental change and socio-cultural adaptation responses to explain non-migratory experiences. Our discussion provides insight into how to best support non-migrant people's adaptive capacity in the face of growing climate emergency.

Evaluating rural participation in wetland management: A contingent valuation analysis of the set-aside policy in Iran.

Growing conditions of water scarcity and population growth necessitate measures for improved water availability to meet agricultural, industrial, and domestic and consumer water demands; generating new environmental pressures on wetlands and other aquatic ecosystems. In Iran, the "set-aside program" incentivizes farmer participation in wetland conservation through mandated land management practices, making them key stakeholders in environmental conservation action. This study explores attitudes to participation in the set-aside wetland conservation program to revive the Jazmurian wetland in Iran, using a random sample of 226 farmer-stakeholder respondents. Farmers were surveyed to investigate economic and social participation using a willingness to accept (WTA) and willingness to pay (in money per ha) (WTP) model. Results show strong (45%) respondent opposition to wetland conservation participation. On the basis of their WTA, the amount of compensation offered by the villagers was significantly affected by "the cultivated area", "gender", "education", "family size", "residency", "income", "moralism", and "Inverse Mills Ratio index" factors. Also, "the cultivated area", "age", "education", "marital status", "family size", and "income" were found to be significantly affecting their WTP. We argue firstly, that policies to improve wetland conservation must join together infrastructure and agricultural development planning - such that dam projects, agricultural and water conservation planning are better integrated across wetland catchments. Secondly, that wetland conservation participation will be improved through land consolidation agreements for small-holders, and through incomes stabilization, capacity building, social learning and awareness-raising initiatives for farmers towards sustainable agricultural practices.

Vancomycin-resistant Enterococcus faecium Empyema in an Asplenic Patient.

Enterococcal empyema is a rare complication of pneumonia. We report the case of a 67-year-old asplenic man with pneumonia complicated by respiratory failure and empyema requiring decortication and prolonged chest tube drainage. Cultures of the empyema were initially negative, but later grew vancomycin-resistant Enterococcus faecium (VRE), which was successfully treated with linezolid. To our knowledge, this is only the second reported case of an empyema caused by VRE that was not associated with an intra-abdominal infection. We suspect superinfection due to airway or chest tube contamination as the most likely mechanism of infection. Physicians should consider multi-drug resistant organisms such as VRE in patients with empyema that fail to resolve with chest tube drainage and broad-spectrum antibiotics.

Intraoperative monitoring of neuromuscular function with soft, skin-mounted wireless devices.

Peripheral nerves are often vulnerable to damage during surgeries, with risks of significant pain, loss of motor function, and reduced quality of life for the patient. Intraoperative methods for monitoring nerve activity are effective, but conventional systems rely on bench-top data acquisition tools with hard-wired connections to electrode leads that must be placed percutaneously inside target muscle tissue. These approaches are time and skill intensive and therefore costly to an extent that precludes their use in many important scenarios. Here we report a soft, skin-mounted monitoring system that measures, stores, and wirelessly transmits electrical signals and physical movement associated with muscle activity, continuously and in real-time during neurosurgical procedures on the peripheral, spinal, and cranial nerves. Surface electromyography and motion measurements can be performed non-invasively in this manner on nearly any muscle location, thereby offering many important advantages in usability and cost, with signal fidelity that matches that of the current clinical standard of care for decision making. These results could significantly improve accessibility of intraoperative monitoring across a broad range of neurosurgical procedures, with associated enhancements in patient outcomes.

Life-Cycle Assessment Harmonization and Soil Science Ranking Results on Food-Waste Management Methods.

This study reviewed 147 life cycle studies, with 28 found suitable for harmonizing food waste management methods' climate and energy impacts. A total of 80 scientific soil productivity studies were assessed to rank management method soil benefits. Harmonized climate impacts per kilogram of food waste range from -0.20 kg of carbon dioxide equivalents (CO2e) for anaerobic digestion (AD) to 0.38 kg of CO2e for landfill gas-to-energy (LFGTE). Aerobic composting (AC) emits -0.10 kg of CO2e. In-sink grinding (ISG) via a food-waste disposer and flushing for management with other sewage at a wastewater treatment plant emits 0.10 kg of CO2e. Harmonization reduced climate emissions versus nonharmonized averages. Harmonized energy impacts range from -0.32 MJ for ISG to 1.14 MJ for AC. AD at 0.27 MJ and LFGTE at 0.40 MJ fall in between. Rankings based on soil studies show AC first for carbon storage and water conservation, with AD second. AD first for fertilizer replacement, with AC second, and AC and AD tied for first for plant yield increase. ISG ranks third and LFGTE fourth on all four soil-quality and productivity indicators. Suggestions for further research include developing soil benefits measurement methods and resolving inconsistencies in the results between life-cycle assessments and soil science studies.

Genetic interrogation of circulating multiple myeloma cells at single-cell resolution.

Multiple myeloma (MM) remains an incurable disease, with a treatment-refractory state eventually developing in all patients. Constant clonal evolution and genetic heterogeneity of MM are a likely explanation for the emergence of drug-resistant disease. Monitoring of MM genomic evolution on therapy by serial bone marrow biopsy is unfortunately impractical because it involves an invasive and painful procedure. We describe how noninvasive and highly sensitive isolation and characterization of circulating tumor cells (CTCs) from peripheral blood at single-cell resolution recapitulate MM in the bone marrow. We demonstrate that CTCs provide the same genetic information as bone marrow MM cells and even reveal mutations with greater sensitivity than bone marrow biopsies in some cases. Single CTC RNA sequencing enables classification of MM and quantitative assessment of genes that are relevant for prognosis. We propose that the genomic characterization of CTCs should be included in clinical trials to follow the emergence of resistant subclones after MM therapy.

An oncogenic MYB feedback loop drives alternate cell fates in adenoid cystic carcinoma.

Translocation events are frequent in cancer and may create chimeric fusions or 'regulatory rearrangements' that drive oncogene overexpression. Here we identify super-enhancer translocations that drive overexpression of the oncogenic transcription factor MYB as a recurrent theme in adenoid cystic carcinoma (ACC). Whole-genome sequencing data and chromatin maps highlight distinct chromosomal rearrangements that juxtapose super-enhancers to the MYB locus. Chromosome conformation capture confirms that the translocated enhancers interact with the MYB promoter. Remarkably, MYB protein binds to the translocated enhancers, creating a positive feedback loop that sustains its expression. MYB also binds enhancers that drive different regulatory programs in alternate cell lineages in ACC, cooperating with TP63 in myoepithelial cells and a Notch program in luminal epithelial cells. Bromodomain inhibitors slow tumor growth in ACC primagraft models in vivo. Thus, our study identifies super-enhancer translocations that drive MYB expression and provides insight into downstream MYB functions in alternate ACC lineages.

A Multiplexed System for Quantitative Comparisons of Chromatin Landscapes.

Genome-wide profiling of histone modifications can provide systematic insight into the regulatory elements and programs engaged in a given cell type. However, conventional chromatin immunoprecipitation and sequencing (ChIP-seq) does not capture quantitative information on histone modification levels, requires large amounts of starting material, and involves tedious processing of each individual sample. Here, we address these limitations with a technology that leverages DNA barcoding to profile chromatin quantitatively and in multiplexed format. We concurrently map relative levels of multiple histone modifications across multiple samples, each comprising as few as a thousand cells. We demonstrate the technology by monitoring dynamic changes following inhibition of p300, EZH2, or KDM5, by linking altered epigenetic landscapes to chromatin regulator mutations, and by mapping active and repressive marks in purified human hematopoietic stem cells. Hence, this technology enables quantitative studies of chromatin state dynamics across rare cell types, genotypes, environmental conditions, and drug treatments.

Th17 cells transdifferentiate into regulatory T cells during resolution of inflammation.

Inflammation is a beneficial host response to infection but can contribute to inflammatory disease if unregulated. The Th17 lineage of T helper (Th) cells can cause severe human inflammatory diseases. These cells exhibit both instability (they can cease to express their signature cytokine, IL-17A) and plasticity (they can start expressing cytokines typical of other lineages) upon in vitro re-stimulation. However, technical limitations have prevented the transcriptional profiling of pre- and post-conversion Th17 cells ex vivo during immune responses. Thus, it is unknown whether Th17 cell plasticity merely reflects change in expression of a few cytokines, or if Th17 cells physiologically undergo global genetic reprogramming driving their conversion from one T helper cell type to another, a process known as transdifferentiation. Furthermore, although Th17 cell instability/plasticity has been associated with pathogenicity, it is unknown whether this could present a therapeutic opportunity, whereby formerly pathogenic Th17 cells could adopt an anti-inflammatory fate. Here we used two new fate-mapping mouse models to track Th17 cells during immune responses to show that CD4(+) T cells that formerly expressed IL-17A go on to acquire an anti-inflammatory phenotype. The transdifferentiation of Th17 into regulatory T cells was illustrated by a change in their signature transcriptional profile and the acquisition of potent regulatory capacity. Comparisons of the transcriptional profiles of pre- and post-conversion Th17 cells also revealed a role for canonical TGF-ß signalling and consequently for the aryl hydrocarbon receptor (AhR) in conversion. Thus, Th17 cells transdifferentiate into regulatory cells, and contribute to the resolution of inflammation. Our data suggest that Th17 cell instability and plasticity is a therapeutic opportunity for inflammatory diseases.

An epigenetic mechanism of resistance to targeted therapy in T cell acute lymphoblastic leukemia.

The identification of activating NOTCH1 mutations in T cell acute lymphoblastic leukemia (T-ALL) led to clinical testing of γ-secretase inhibitors (GSIs) that prevent NOTCH1 activation. However, responses to these inhibitors have been transient, suggesting that resistance limits their clinical efficacy. Here we modeled T-ALL resistance, identifying GSI-tolerant 'persister' cells that expand in the absence of NOTCH1 signaling. Rare persisters are already present in naive T-ALL populations, and the reversibility of their phenotype suggests an epigenetic mechanism. Relative to GSI-sensitive cells, persister cells activate distinct signaling and transcriptional programs and exhibit chromatin compaction. A knockdown screen identified chromatin regulators essential for persister viability, including BRD4. BRD4 binds enhancers near critical T-ALL genes, including MYC and BCL2. The BRD4 inhibitor JQ1 downregulates expression of these targets and induces growth arrest and apoptosis in persister cells, at doses well tolerated by GSI-sensitive cells. Consistently, the GSI-JQ1 combination was found to be effective against primary human leukemias in vivo. Our findings establish a role for epigenetic heterogeneity in leukemia resistance that may be addressed by incorporating epigenetic modulators in combination therapy.

Alternative anterior reference sites for measuring posterior tibial nerve somatosensory evoked potentials.

OBJECTIVE: The purpose of this study was to examine the utility and feasibility of using alternative anterior reference leads when measuring left posterior tibial nerve somatosensory evoked potentials (SEPs). METHODS: With IRB approval, 12 patients were monitored using both traditional (FPz and C4') and alternative anterior (F3 and F4) reference leads during routine spine surgery with SEP monitoring. Recordings from the routine and novel electrode pairs were collected and analyzed. RESULTS: All of the SEP amplitudes measured were of similar magnitude except for that of F3-F4, which was significantly lower (P < 0.001) than all of the other five lead combinations which were assessed (Cz'-FPz, C3'-C4', C3'-F4, Cz'-F3, and Cz'-F4). The latencies of the novel lead combinations (C3'-F4, Cz'-F3, Cz'-F4, and F3-F4) were similar to those of the "gold standards" (Cz'-FPz and C3'-C4') (pooled median, 45.6 ms with 25-75th percentiles, 44.0-47.8 ms, P = 0.308). The coefficients of variation (CV %) of the amplitudes were not statistically significantly different (P = 0.341). CONCLUSIONS: The use of alternative frontal reference leads (F3 and F4) for left posterior tibial nerve SEP monitoring yields signals of equal quality and reproducibility compared to signals with standard (FPz and C4') referencing. These alternative leads may substitute for traditional referencing when placement of FPz or C4' is precluded by the location of surgery.