BET inhibition reforms the immune microenvironment and alleviates T cell dysfunction in chronic lymphocytic leukemia.

Redundant tumor microenvironment (TME) immunosuppressive mechanisms and epigenetic maintenance of terminal T cell exhaustion greatly hinder functional antitumor immune responses in chronic lymphocytic leukemia (CLL). Bromodomain and extraterminal (BET) proteins regulate key pathways contributing to CLL pathogenesis and TME interactions, including T cell function and differentiation. Herein, we report that blocking BET protein function alleviates immunosuppressive networks in the CLL TME and repairs inherent CLL T cell defects. The pan-BET inhibitor OPN-51107 reduced exhaustion-associated cell signatures resulting in improved T cell proliferation and effector function in the Eµ-TCL1 splenic TME. Following BET inhibition (BET-i), TME T cells coexpressed significantly fewer inhibitory receptors (IRs) (e.g., PD-1, CD160, CD244, LAG3, VISTA). Complementary results were witnessed in primary CLL cultures, wherein OPN-51107 exerted proinflammatory effects on T cells, regardless of leukemic cell burden. BET-i additionally promotes a progenitor T cell phenotype through reduced expression of transcription factors that maintain terminal differentiation and increased expression of TCF-1, at least in part through altered chromatin accessibility. Moreover, direct T cell effects of BET-i were unmatched by common targeted therapies in CLL. This study demonstrates the immunomodulatory action of BET-i on CLL T cells and supports the inclusion of BET inhibitors in the management of CLL to alleviate terminal T cell dysfunction and potentially enhance tumoricidal T cell activity.

Local gate control of Mott metal-insulator transition in a 2D metal-organic framework.

Electron-electron interactions in materials lead to exotic many-body quantum phenomena, including Mott metal-insulator transitions (MITs), magnetism, quantum spin liquids, and superconductivity. These phases depend on electronic band occupation and can be controlled via the chemical potential. Flat bands in two-dimensional (2D) and layered materials with a kagome lattice enhance electronic correlations. Although theoretically predicted, correlated-electron Mott insulating phases in monolayer 2D metal-organic frameworks (MOFs) with a kagome structure have not yet been realised experimentally. Here, we synthesise a 2D kagome MOF on a 2D insulator. Scanning tunnelling microscopy (STM) and spectroscopy reveal a MOF electronic energy gap of ∼200 meV, consistent with dynamical mean-field theory predictions of a Mott insulator. Combining template-induced (via work function variations of the substrate) and STM probe-induced gating, we locally tune the electron population of the MOF kagome bands and induce Mott MITs. These findings enable technologies based on electrostatic control of many-body quantum phases in 2D MOFs.

Inhibition of Bromodomain and Extra Terminal (BET) Domain Activity Modulates the IL-23R/IL-17 Axis and Suppresses Acute Graft-Versus-Host Disease.

Acute graft-versus-host disease (GVHD) is the leading cause of non-relapse mortality following allogeneic hematopoietic cell transplantation. The majority of patients non-responsive to front line treatment with steroids have an estimated overall 2-year survival rate of only 10%. Bromodomain and extra-terminal domain (BET) proteins influence inflammatory gene transcription, and therefore represent a potential target to mitigate inflammation central to acute GVHD pathogenesis. Using potent and selective BET inhibitors Plexxikon-51107 and -2853 (PLX51107 and PLX2853), we show that BET inhibition significantly improves survival and reduces disease progression in murine models of acute GVHD without sacrificing the beneficial graft-versus-leukemia response. BET inhibition reduces T cell alloreactive proliferation, decreases inflammatory cytokine production, and impairs dendritic cell maturation both in vitro and in vivo. RNA sequencing studies in human T cells revealed that BET inhibition impacts inflammatory IL-17 and IL-12 gene expression signatures, and Chromatin Immunoprecipitation (ChIP)-sequencing revealed that BRD4 binds directly to the IL-23R gene locus. BET inhibition results in decreased IL-23R expression and function as demonstrated by decreased phosphorylation of STAT3 in response to IL-23 stimulation in human T cells in vitro as well as in mouse donor T cells in vivo. Furthermore, PLX2853 significantly reduced IL-23R+ and pathogenic CD4+ IFNγ+ IL-17+ double positive T cell infiltration in gastrointestinal tissues in an acute GVHD murine model. Our findings identify a role for BET proteins in regulating the IL-23R/STAT3/IL-17 pathway. Based on our preclinical data presented here, PLX51107 will enter clinical trial for refractory acute GVHD in a Phase 1 safety, biological efficacy trial.

Variable bites and dynamic populations; new insights in Leishmania transmission.

Leishmaniasis is a neglected tropical disease which kills an estimated 50,000 people each year, with its deadly impact confined mainly to lower to middle income countries. Leishmania parasites are transmitted to human hosts by sand fly vectors during blood feeding. Recent experimental work shows that transmission is modulated by the patchy landscape of infection in the host's skin, and the parasite population dynamics within the vector. Here we assimilate these new findings into a simple probabilistic model for disease transmission which replicates recent experimental results, and assesses their relative importance. The results of subsequent simulations, describing random parasite uptake and dynamics across multiple blood meals, show that skin heterogeneity is important for transmission by short-lived flies, but that for longer-lived flies with multiple bites the population dynamics within the vector dominate transmission probability. Our results indicate that efforts to reduce fly lifespan beneath a threshold of around two weeks may be especially helpful in reducing disease transmission.

Fast, accurate enthalpy differences in spin crossover crystals from DFT+U.

Spin crossover materials are bi-stable systems with potential applications as molecular scale electronic switches, actuators, thermometers, barometers, and displays. However, calculating the enthalpy difference, ΔH, between the high spin and low spin states has been plagued with difficulties. For example, many common density functional theory (DFT) methods fail to even predict the correct sign of ΔH, which determines the low temperature state. Here, we study a collection of Fe(II) and Fe(III) materials, where ΔH has been measured, which has previously been used to benchmark density functionals. The best performing hybrid functional, TPSSh, achieves a mean absolute error compared to experiment of 11 kJ mol-1 for this set of materials. However, hybrid functionals scale badly in the solid state; therefore, local functionals are preferable for studying crystalline materials, where the most interesting spin crossover phenomena occur. We show that both the Liechtenstein and Dudarev DFT+U methods are a little more accurate than TPSSh. The Dudarev method yields a mean absolute error of 8 kJ mol-1 for Ueff = 1.6 eV. However, the mean absolute error for both TPSSh and DFT+U is dominated by a single material, for which the two theoretical methods predict similar enthalpy differences-if this is excluded from the set, then DFT+U achieves chemical accuracy. Thus, DFT+U is an attractive option for calculating the properties of spin crossover crystals, as its accuracy is comparable to that of meta-hybrid functionals, but at a much lower computational cost.

BET inhibitors synergize with venetoclax to induce apoptosis in MYC-driven lymphomas with high BCL-2 expression.

Although the MYC oncogenic network represents an attractive therapeutic target for lymphoma, MYC inhibitors have been difficult to develop. Alternatively, inhibitors of epigenetic/ transcriptional regulators, particularly the bromodomain and extraterminal (BET) family, have been used to modulate MYC. However, current benzodiazepine-derivative BET inhibitors (BETi) elicit disappointing responses and dose-limiting toxicity in relapsed/refractory lymphoma, potentially because of enrichment of high-risk molecular features and chemical backbone-associated toxicities. Consequently, novel nonbenzodiazepine BETi and improved mechanistic understanding are required. Here we characterize the responses of aggressive MYC-driven lymphomas to 2 nonbenzodiazepine BETi: PLX51107 and PLX2853. Both invoked BIM-dependent apoptosis and in vivo therapy, associated with miR-17∼92 repression, in murine Eµ-myc lymphomas, with PLX2853 exhibiting enhanced potency. Accordingly, exogenous BCL-2 expression abrogated these effects. Because high BCL-2 expression is common in diffuse large B-cell lymphoma (DLBCL), BETi were ineffective in driving apoptosis and in vivo therapy of DLBCL cell lines, mirroring clinical results. However, BETi-mediated BIM upregulation and miR-17∼92 repression remained intact. Consequently, coadministration of BETi and ABT199/venetoclax restored cell death and in vivo therapy. Collectively, these data identify BIM-dependent apoptosis as a critical mechanism of action for this class of BETi that, via coadministration of BH3 mimetics, can deliver effective tumor control in DLBCL.

Mechanomagnetics in Elastic Crystals: Insights from [Cu(acac)2 ].

We predict that the magnetic properties of [Cu(acac)2 ], an elastically flexible crystal, change drastically when the crystal is bent. It is found that unbent [Cu(acac)2 ] is an almost perfect Tomonaga-Luttinger liquid. Broken-symmetry density-functional calculations reveal that the magnetic exchange interactions along the chains are an order of magnitude larger than the interchain exchange. The geometrically frustrated interchain interactions cannot magnetically order the material at any experimentally accessible temperature. The ordering temperature (TN ), calculated from the chain-random-phase approximation, increases by 24 orders of magnitude when the material is bent. We demonstrate that geometric frustration both suppresses TN and enhances the sensitivity of TN to bending. In [Cu(acac)2 ], TN is extremely sensitive to bending but remains too low for practical applications, even when bent. Partially frustrated materials could achieve the balance of high TN and good sensitivity to bending required for practical applications of mechanomagnetic elastic crystals.

BRD4 Profiling Identifies Critical Chronic Lymphocytic Leukemia Oncogenic Circuits and Reveals Sensitivity to PLX51107, a Novel Structurally Distinct BET Inhibitor.

Bromodomain and extra-terminal (BET) family proteins are key regulators of gene expression in cancer. Herein, we utilize BRD4 profiling to identify critical pathways involved in pathogenesis of chronic lymphocytic leukemia (CLL). BRD4 is overexpressed in CLL and is enriched proximal to genes upregulated or de novo expressed in CLL with known functions in disease pathogenesis and progression. These genes, including key members of the B-cell receptor (BCR) signaling pathway, provide a rationale for this therapeutic approach to identify new targets in alternative types of cancer. Additionally, we describe PLX51107, a structurally distinct BET inhibitor with novel in vitro and in vivo pharmacologic properties that emulates or exceeds the efficacy of BCR signaling agents in preclinical models of CLL. Herein, the discovery of the involvement of BRD4 in the core CLL transcriptional program provides a compelling rationale for clinical investigation of PLX51107 as epigenetic therapy in CLL and application of BRD4 profiling in other cancers.Significance: To date, functional studies of BRD4 in CLL are lacking. Through integrated genomic, functional, and pharmacologic analyses, we uncover the existence of BRD4-regulated core CLL transcriptional programs and present preclinical proof-of-concept studies validating BET inhibition as an epigenetic approach to target BCR signaling in CLL. Cancer Discov; 8(4); 458-77. ©2018 AACR.This article is highlighted in the In This Issue feature, p. 371.

Physical and chemical impacts of a major storm on a temperate lake: a taste of things to come?

Extreme weather can have a substantial influence on lakes and is expected to become more frequent with climate change. We explored the influence of one particular extreme event, Storm Ophelia, on the physical and chemical environment of England's largest lake, Windermere. We found that the substantial influence of Ophelia on meteorological conditions at Windermere, in particular wind speed, resulted in a 25-fold increase (relative to the study-period average) in the wind energy flux at the lake-air interface. Following Ophelia, there was a short-lived mixing event in which the Schmidt stability decreased by over 100 Jm-2 and the thermocline deepened by over 10 m during a 12-h period. As a result of changes to the strength of stratification, Ophelia also changed the internal seiche regime of Windermere with the dominant seiche period increasing from ~ 17 h pre-storm to ~ 21 h post-storm. Following Ophelia, there was an upwelling of cold and low-oxygenated waters at the southern-end of the lake. This had a substantial influence on the main outflow of Windermere, the River Leven, where dissolved oxygen concentrations decreased by ~ 48%, from 9.3 to 4.8 mg L-1, while at the mid-lake monitoring station in Windermere, it decreased by only ~ 3%. This study illustrates that the response of a lake to extreme weather can cause important effects downstream, the influence of which may not be evident at the lake surface. To understand the impact of future extreme events fully, the whole lake and downstream-river system need to be studied together.

Current Challenges in Glioblastoma: Intratumour Heterogeneity, Residual Disease, and Models to Predict Disease Recurrence.

Glioblastoma (GB) is the most common primary malignant brain tumor, and despite the availability of chemotherapy and radiotherapy to combat the disease, overall survival remains low with a high incidence of tumor recurrence. Technological advances are continually improving our understanding of the disease, and in particular, our knowledge of clonal evolution, intratumor heterogeneity, and possible reservoirs of residual disease. These may inform how we approach clinical treatment and recurrence in GB. Mathematical modeling (including neural networks) and strategies such as multiple sampling during tumor resection and genetic analysis of circulating cancer cells, may be of great future benefit to help predict the nature of residual disease and resistance to standard and molecular therapies in GB.

RAF inhibitors that evade paradoxical MAPK pathway activation.

Oncogenic activation of BRAF fuels cancer growth by constitutively promoting RAS-independent mitogen-activated protein kinase (MAPK) pathway signalling. Accordingly, RAF inhibitors have brought substantially improved personalized treatment of metastatic melanoma. However, these targeted agents have also revealed an unexpected consequence: stimulated growth of certain cancers. Structurally diverse ATP-competitive RAF inhibitors can either inhibit or paradoxically activate the MAPK pathway, depending whether activation is by BRAF mutation or by an upstream event, such as RAS mutation or receptor tyrosine kinase activation. Here we have identified next-generation RAF inhibitors (dubbed 'paradox breakers') that suppress mutant BRAF cells without activating the MAPK pathway in cells bearing upstream activation. In cells that express the same HRAS mutation prevalent in squamous tumours from patients treated with RAF inhibitors, the first-generation RAF inhibitor vemurafenib stimulated in vitro and in vivo growth and induced expression of MAPK pathway response genes; by contrast the paradox breakers PLX7904 and PLX8394 had no effect. Paradox breakers also overcame several known mechanisms of resistance to first-generation RAF inhibitors. Dissociating MAPK pathway inhibition from paradoxical activation might yield both improved safety and more durable efficacy than first-generation RAF inhibitors, a concept currently undergoing human clinical evaluation with PLX8394.

Characterizing and Overriding the Structural Mechanism of the Quizartinib-Resistant FLT3 "Gatekeeper" F691L Mutation with PLX3397.

UNLABELLED: Tyrosine kinase domain mutations are a common cause of acquired clinical resistance to tyrosine kinase inhibitors (TKI) used to treat cancer, including the FLT3 inhibitor quizartinib. Mutation of kinase "gatekeeper" residues, which control access to an allosteric pocket adjacent to the ATP-binding site, has been frequently implicated in TKI resistance. The molecular underpinnings of gatekeeper mutation-mediated resistance are incompletely understood. We report the first cocrystal structure of FLT3 with the TKI quizartinib, which demonstrates that quizartinib binding relies on essential edge-to-face aromatic interactions with the gatekeeper F691 residue, and F830 within the highly conserved Asp-Phe-Gly motif in the activation loop. This reliance makes quizartinib critically vulnerable to gatekeeper and activation loop substitutions while minimizing the impact of mutations elsewhere. Moreover, we identify PLX3397, a novel FLT3 inhibitor that retains activity against the F691L mutant due to a binding mode that depends less vitally on specific interactions with the gatekeeper position. SIGNIFICANCE: We report the first cocrystal structure of FLT3 with a kinase inhibitor, elucidating the structural mechanism of resistance due to the gatekeeper F691L mutation. PLX3397 is a novel FLT3 inhibitor with in vitro activity against this mutation but is vulnerable to kinase domain mutations in the FLT3 activation loop.

Preliminary report of a sutureless onlay technique for incisional hernia repair using fibrin glue alone for mesh fixation.

The Rives repair for ventral/incisional (V/I) hernias involves sublay mesh placement requiring retrorectus dissection and transfascial stitches. Chevrel described a repair by onlaying mesh after a unique primary fascial closure. Although Chevrel fixated mesh to the anterior fascia with sutures, he used fibrin glue for fascial closure reinforcement. We describe an onlay technique with mesh fixated to the anterior fascia solely with fibrin glue without suture fixation. From January 2010 to January 2012, 50 patients underwent a V/I hernia onlay technique with fibrin glue mesh fixation. Records were reviewed for technical details, demographics, mesh characteristics, and postoperative outcomes. Primary fascial closure with interrupted permanent suture was done with or without myofascial advancement flaps. Onlay polypropylene mesh was placed providing 8 cm of overlap. Fibrin glue was applied over the prosthesis and subcutaneous drains were placed. Mean age was 62.4 years. Mean body mass index was 30.1 kg/m(2). Average mesh size was 14.5 cm × 19.1 cm. Mean operative time was 144.4 minutes (range, 38 to 316 minutes). Mean discharge was postoperative Day 2.9 (range, 0 to 15 days). Morbidity included eight seromas, one hematoma, and three wound infections. Seventeen patients required components separation. Mean follow-up was 19.5 months with no recurrences. This is the first series describing fibrin glue alone for mesh fixation for V/I hernia repair. It allows for immediate prosthesis fixation to the anterior fascia. Early results are promising. Potential advantages include less operative time, less technical difficulty, and less long-term pain. A prospective trial is needed to evaluate this approach.

Design and pharmacology of a highly specific dual FMS and KIT kinase inhibitor.

Inflammation and cancer, two therapeutic areas historically addressed by separate drug discovery efforts, are now coupled in treatment approaches by a growing understanding of the dynamic molecular dialogues between immune and cancer cells. Agents that target specific compartments of the immune system, therefore, not only bring new disease modifying modalities to inflammatory diseases, but also offer a new avenue to cancer therapy by disrupting immune components of the microenvironment that foster tumor growth, progression, immune evasion, and treatment resistance. McDonough feline sarcoma viral (v-fms) oncogene homolog (FMS) and v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog (KIT) are two hematopoietic cell surface receptors that regulate the development and function of macrophages and mast cells, respectively. We disclose a highly specific dual FMS and KIT kinase inhibitor developed from a multifaceted chemical scaffold. As expected, this inhibitor blocks the activation of macrophages, osteoclasts, and mast cells controlled by these two receptors. More importantly, the dual FMS and KIT inhibition profile has translated into a combination of benefits in preclinical disease models of inflammation and cancer.

Teacher efficacy and pupil behaviour: the structure of teachers' individual and collective beliefs and their relationship with numbers of pupils excluded from school.

BACKGROUND: Previous work has yielded knowledge of teachers' attributions for children's behaviour. Other studies have helped to develop understanding of teachers' efficacy beliefs. Little work has been undertaken to examine teachers' efficacy beliefs with regard to classroom behaviour. AIMS: This study aimed to investigate the relationship between teachers' individual and collective beliefs about their efficacy with children's behaviour and whether these beliefs were associated with the use of exclusion as a sanction. SAMPLE: A total of 197 teachers from 31 primary and nursery schools in the North East of England participated. METHODS: Participants responded to questionnaires to assess their individual and collective efficacy beliefs. Demographic and school level data were also collected. RESULTS: Factor analysis indicated that teachers' individual efficacy beliefs were best represented by three factors: 'Classroom Management', 'Children's Engagement', 'Instructional Strategies' that corresponded well to previous findings. Analysis of collective efficacy beliefs showed a similar structure that differed from previous findings. Individual efficacy was not associated with numbers of children excluded. One factor 'Addressing External Influences' in the collective beliefs was negatively correlated with numbers of children excluded and appeared to mitigate the deleterious effects associated with socio-economic deprivation. CONCLUSIONS: This study adds weight to the importance of understanding and supporting teachers' beliefs in their collective efficacy. In particular, this study underlines the need for strategies that will endorse and develop teachers' beliefs in their ability to manage children's behaviour successfully.

ATPase cycle and DNA unwinding kinetics of RecG helicase.

The superfamily 2 bacterial helicase, RecG, is a monomeric enzyme with a role in DNA repair by reversing stalled replication forks. The helicase must act specifically and rapidly to prevent replication fork collapse. We have shown that RecG binds tightly and rapidly to four-strand oligonucleotide junctions, which mimic a stalled replication fork. The helicase unwinds such DNA junctions with a step-size of approximately four bases per ATP hydrolyzed. To gain an insight into this mechanism, we used fluorescent stopped-flow and quenched-flow to measure individual steps within the ATPase cycle of RecG, when bound to a DNA junction. The fluorescent ATP analogue, mantATP, was used throughout to determine the rate limiting steps, effects due to DNA and the main states in the cycle. Measurements, when possible, were also performed with unlabeled ATP to confirm the mechanism. The data show that the chemical step of hydrolysis is the rate limiting step in the cycle and that this step is greatly accelerated by bound DNA. The ADP release rate is similar to the cleavage rate, so that bound ATP and ADP would be the main states during the ATP cycle. Evidence is provided that the main structural rearrangements, which bring about DNA unwinding, are linked to these states.

Gastric diverticulum: "a wayside house of ill fame" with a laparoscopic solution.

BACKGROUND: Gastric diverticulum (GD) is an extremely rare disorder that can easily be overlooked when investigating the cause of abdominal pain. Its diagnosis is founded on a history of gastrointestinal symptoms and a typically unrevealing physical examination, and diagnosis requires confirmation from UGI contrast studies, EGD, and CT scan. Symptomatic GD should be kept in consideration as a cause of abdominal issues, because not only is it treatable, but also complications of GD can be life threatening. The surgical treatment of GDs has evolved from thoraco-abdominal incisions in the early twentieth century to the laparoscopic approach used today. CASE REPORT: The patient is a 45-y-old male presenting with a 4-mo case of dysphagia, small amounts of regurgitation, and abdominal pain but no other symptoms. RESULTS: The patient was diagnosed with a gastric diverticulum, which was subsequently successfully treated with a laparoscopic gastric diverticulectomy. CONCLUSION: Laparoscopic gastric diverticulectomy is a safe procedure and should be considered as an option to treat symptomatic GD.