Epigenome-wide analysis of T-cell large granular lymphocytic leukemia identifies BCL11B as a potential biomarker

Background: The molecular pathogenesis of T-cell large granular lymphocytic leukemia (T-LGLL), a mature T-cell leukemia arising commonly from T-cell receptor alpha beta-positive CD8(+) memory cytotoxic T cells, is only partly understood. The role of deregulated methylation in T-LGLL is not well known. We analyzed the epigenetic profile of T-LGLL cells of 11 patients compared to their normal counterparts by array-based DNA methylation profiling. For identification of molecular events driving the pathogenesis of T-LGLL, we compared the differentially methylated loci between the T-LGLL cases and normal T cells with chromatin segmentation data of benign T cells from the BLUEPRINT project. Moreover, we analyzed gene expression data of T-LGLL and benign T cells and validated the results by pyrosequencing in an extended cohort of 17 patients, including five patients with sequential samples. Results: We identified dysregulation of DNA methylation associated with altered gene expression in T-LGLL. Since T-LGLL is a rare disease, the samples size is low. But as confirmed for each sample, hypermethylation of T-LGLL cells at various CpG sites located at enhancer regions is a hallmark of this disease. The interaction of BLC11B and C14orf64 as suggested by in silico data analysis could provide a novel pathogenetic mechanism that needs further experimental investigation. Conclusions: DNA methylation is altered in T-LGLL cells compared to benign T cells. In particular, BCL11B is highly significant differentially methylated in T-LGLL cells. Although our results have to be validated in a larger patient cohort, BCL11B could be considered as a potential biomarker for this leukemia. In addition, altered gene expression and hypermethylation of enhancer regions could serve as potential mechanisms for treatment of this disease. Gene interactions of dysregulated genes, like BLC11B and C14orf64, may play an important role in pathogenic mechanisms and should be further analyzed

European consensus statement on essential colposcopy

Peer reviewedPostprin

Inflammation and tissue repair markers distinguish the nodular sclerosis and mixed cellularity subtypes of classical Hodgkin's lymphoma

Background: Classical Hodgkin's lymphoma (cHL), although a malignant disease, has many features in common with an inflammatory condition. The aim of this study was to establish the molecular characteristics of the two most common cHL subtypes, nodular sclerosis (NS) and mixed cellularity (MC), based on molecular profiling and immunohistochemistry, with special reference to the inflammatory microenvironment. Methods: We analysed 44 gene expression profiles of cHL whole tumour tissues, 25 cases of NS and 19 cases of MC, using Affymetrix chip technology and immunohistochemistry. Results: In the NS subtype, 152 genes showed a significantly higher expression, including genes involved in extracellular matrix (ECM) remodelling and ECM deposition similar to wound healing. Among these were SPARC, CTSK and COLI. Immunohistochemistry revealed that the NS-related genes were mainly expressed by macrophages and fibroblasts. Fifty-three genes had a higher expression in the MC subtype, including several inflammation-related genes, such as C1Qα, C1Qβ and CXCL9. In MC tissues, the C1Q subunits were mainly expressed by infiltrating macrophages. Conclusions and interpretations: We suggest that the identified subtype-specific genes could reflect different phases of wound healing. Our study underlines the potential function of infiltrating macrophages in shaping the cHL tumour microenvironment

The JAK inhibitor AZD1480 regulates proliferation and immunity in Hodgkin lymphoma

Aberrant activation of the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway has been reported to promote proliferation and survival of Hodgkin and Reed–Sternberg cells of Hodgkin lymphoma (HL). We investigated the activity of the JAK inhibitor AZD1480 in HL-derived cell lines and determined its mechanisms of action. AZD1480 at low doses (0.1–1 μ) potently inhibited STATs phosphorylation, but did not predictably result in antiproliferative effects, as it activated a negative-feedback loop causing phosphorylation of JAK2 and extracellular signal-regulated kinases 1 and 2 (ERK1/2), and increased IP-10, RANTES and interleukin (IL)-8 concentrations in the supernatants. Inhibition of the ERK activity by mitogen-activated extracellular signal regulated kinase (MEK) inhibitors (UO126 and PD98059) enhanced the cytotoxic activity of AZD1480. Interestingly, submicromolar concentrations of AZD1480 demonstrated significant immunoregulatory effects by downregulating T-helper 2 cytokines and chemokines, including IL-13 and thymus- and activation-regulated chemokine, and the surface expression of the immunosuppressive programmed death ligands 1 and 2. Higher concentrations of AZD1480 (5 μ) induced G2/M arrest and cell death by inhibiting Aurora kinases. Our study demonstrates that AZD1480 regulates proliferation and immunity in HL cell lines and provides mechanistic rationale for evaluating AZD1480 alone or in combination with MEK inhibitors in HL

VEGFR2 pY949 signalling regulates adherens junction integrity and metastatic spread

The specific role of VEGFA-induced permeability and vascular leakage in physiology and pathology has remained unclear. Here we show that VEGFA-induced vascular leakage depends on signalling initiated via the VEGFR2 phosphosite Y949, regulating dynamic c-Src and VE-cadherin phosphorylation. Abolished Y949 signalling in the mouse mutant Vegfr2Y949F/Y949F leads to VEGFA-resistant endothelial adherens junctions and a block in molecular extravasation. Vessels in Vegfr2Y949F/Y949F mice remain sensitive to inflammatory cytokines, and vascular morphology, blood pressure and flow parameters are normal. Tumour-bearing Vegfr2Y949F/Y949F mice display reduced vascular leakage and oedema, improved response to chemotherapy and, importantly, reduced metastatic spread. The inflammatory infiltration in the tumour micro-environment is unaffected. Blocking VEGFAinduced disassembly of endothelial junctions, thereby suppressing tumour oedema and metastatic spread, may be preferable to full vascular suppression in the treatment of certain cancer forms

Diverse Hematological Malignancies Including Hodgkin-Like Lymphomas Develop in Chimeric MHC Class II Transgenic Mice

A chimeric HLA-DR4-H2-E (DR4) homozygous transgenic mouse line spontaneously develops diverse hematological malignancies with high frequency (70%). The majority of malignancies were distributed equally between T and B cell neoplasms and included lymphoblastic T cell lymphoma (LTCL), lymphoblastic B cell lymphoma (LBCL), diffuse large B cell lymphoma (DLBCL), the histiocyte/T cell rich variant of DLBCL (DLBCL-HA/T cell rich DLBCL), splenic marginal zone lymphoma (SMZL), follicular B cell lymphoma (FBL) and plasmacytoma (PCT). Most of these neoplasms were highly similar to human diseases. Also, some non-lymphoid malignancies such as acute myeloid leukemia (AML) and histiocytic sarcoma were found. Interestingly, composite lymphomas, including Hodgkin-like lymphomas, were also detected that had CD30+ Hodgkin/Reed-Sternberg (H/RS)-like cells, representing a tumor type not previously described in mice. Analysis of microdissected H/RS-like cells revealed their origin as germinal center B cells bearing somatic hypermutations and, in some instances, crippled mutations, as described for human Hodgkin lymphoma (HL). Transgene integration in an oncogene was excluded as an exclusive driving force of tumorigenesis and age-related lymphoma development suggests a multi-step process. Thus, this DR4 line is a useful model to investigate common molecular mechanisms that may contribute to important neoplastic diseases in man

Comet 67P/Churyumov-Gerasimenko: Constraints on its origin from OSIRIS observations

Context. One of the main aims of the ESA Rosetta mission is to study the origin of the solar system by exploring comet 67P/Churyumov-Gerasimenko at close range. Aims. In this paper we discuss the origin and evolution of comet 67P/Churyumov-Gerasimenko in relation to that of comets in general and in the framework of current solar system formation models. Methods. We use data from the OSIRIS scientific cameras as basic constraints. In particular, we discuss the overall bi-lobate shape and the presence of key geological features, such as layers and fractures. We also treat the problem of collisional evolution of comet nuclei by a particle-in-a-box calculation for an estimate of the probability of survival for 67P/Churyumov-Gerasimenko during the early epochs of the solar system. Results. We argue that the two lobes of the 67P/Churyumov-Gerasimenko nucleus are derived from two distinct objects that have formed a contact binary via a gentle merger. The lobes are separate bodies, though sufficiently similar to have formed in the same environment. An estimate of the collisional rate in the primordial, trans-planetary disk shows that most comets of similar size to 67P/Churyumov-Gerasimenko are likely collisional fragments, although survival of primordial planetesimals cannot be excluded. Conclusions. A collisional origin of the contact binary is suggested, and the low bulk density of the aggregate and abundance of volatile species show that a very gentle merger must have occurred. We thus consider two main scenarios: the primordial accretion of planetesimals, and the re-accretion of fragments after an energetic impact onto a larger parent body. We point to the primordial signatures exhibited by 67P/Churyumov-Gerasimenko and other comet nuclei as critical tests of the collisional evolution

Characterization of OSIRIS NAC filters for the interpretation of multispectral data of comet 67P/Churyumov-Gerasimenko

We interpret multicolor data from OSIRISNAC for the remote-sensing exploration of comet 67P/Churyumov-Gerasimenko. We determine the most meaningful definition of color maps for the characterization of surface variegation with filters available on OSIRIS NAC. Methods. We analyzed laboratory spectra of selected minerals and olivine-pyroxene mixtures seen through OSIRIS NAC filters, with spectral methods existing in the literature: reflectance ratios, minimum band wavelength, spectral slopes, band tilt, band curvature, and visible tilt. Results. We emphasize the importance of reflectance ratios and particularly the relation of visible tilt vs. band tilt. This technique provides a reliable diagnostic of the presence of silicates. Color maps constructed by red-green-blue colors defined with the green, orange, red, IR, and Fe2O3 filters let us define regions that may significantly differ in composition

Acceleration of individual, decimetre-sized aggregates in the lower coma of comet 67P/Churyumov-Gerasimenko

We present observations of decimetre-sized, likely ice-containing aggregates ejected from a confined region on the surface of comet 67P/Churyumov-Gerasimenko. The images were obtained with the narrow angle camera of the Optical, Spectroscopic, and Infrared Remote Imaging System on board the Rosetta spacecraft in 2016 January when the comet was at 2 au from the Sun outbound from perihelion.We measure the acceleration of individual aggregates through a 2 h image series. Approximately 50 per cent of the aggregates are accelerated away from the nucleus, and 50 per cent towards it, and likewise towards either horizontal direction. The accelerations are up to one order of magnitude stronger than local gravity, and are most simply explained by the combined effect of gas drag accelerating all aggregates upwards, and the recoil force from asymmetric outgassing, either from rotating aggregates with randomly oriented spin axes and sufficient thermal inertia to shift the temperature maximum away from an aggregate's subsolar region, or from aggregates with variable ice content. At least 10 per cent of the aggregates will escape the gravity field of the nucleus and feed the comet's debris trail, while others may fall back to the surface and contribute to the deposits covering parts of the Northern hemisphere. The rocket force plays a crucial role in pushing these aggregates back towards the surface. Our observations show the future back fall material in the process of ejection, and provide the first direct measurement of the acceleration of aggregates in the innermost coma (<2 km) of a comet, where gas drag is still significant

A mini outburst from the nightside of comet 67P/Churyumov-Gerasimenko observed by the OSIRIS camera on Rosetta

Context. On 12 March 2015 the OSIRIS WAC camera onboard the ESA Rosetta spacecraft orbiting comet 67P/Churyumov-Gerasimenko observed a small outburst originating from the Imhotep region at the foot of the big lobe of the comet. These measurements are unique since it was the first time that the initial phase of a transient outburst event could be directly observed. Aims. We investigate the evolution of the dust jet in order to derive clues about the outburst source mechanism and the ejected dust particles, in particular the dust mass, dust-to-gas ratio and the particle size distribution. Methods. Analysis of the images and of the observation geometry using comet shape models in combination with gasdynamic modeling of the transient dust jet were the main tools used in this study. Synthetic images were computed for comparison with the observations. Results. Analysis of the geometry revealed that the source region was not illuminated until 1.5 h after the event implying true nightside activity was observed. The outburst lasted for less than one hour and the average dust production rate during the initial four minutes was of the order of 1 kg/s. During this time the outburst dust production rate was approximately constant, no sign for an initial explosion could be detected. For dust grains between 0.01-1 mm a power law size distribution characterized by an index of about 2.6 provides the best fit to the observed radiance profiles. The dust-to-gas ratio of the outburst jet is in the range 0.6-1.8