A genome-wide search for epigenetically regulated genes in zebra finch using MethylCap-seq and RNA-seq

Learning and memory formation are known to require dynamic CpG (de)methylation and gene expression changes. Here, we aimed at establishing a genome-wide DNA methylation map of the zebra finch genome, a model organism in neuroscience, as well as identifying putatively epigenetically regulated genes. RNA-and MethylCap-seq experiments were performed on two zebra finch cell lines in presence or absence of 5-aza-2'-deoxycytidine induced demethylation. First, the MethylCap-seq methodology was validated in zebra finch by comparison with RRBS-generated data. To assess the influence of (variable) methylation on gene expression, RNA-seq experiments were performed as well. Comparison of RNA-seq and MethylCap-seq results showed that at least 357 of the 3,457 AZA-upregulated genes are putatively regulated by methylation in the promoter region, for which a pathway analysis showed remarkable enrichment for neurological networks. A subset of genes was validated using Exon Arrays, quantitative RT-PCR and CpG pyrosequencing on bisulfite-treated samples. To our knowledge, this study provides the first genome-wide DNA methylation map of the zebra finch genome as well as a comprehensive set of genes of which transcription is under putative methylation control

High-fidelity CRISPR/Cas9-based gene-specific hydroxymethylation rescues gene expression and attenuates renal fibrosis

While suppression of specific genes through aberrant promoter methylation contributes to different diseases including organ fibrosis, gene-specific reactivation technology is not yet available for therapy. TET enzymes catalyze hydroxymethylation of methylated DNA, reactivating gene expression. We here report generation of a high-fidelity CRISPR/Cas9-based gene-specific dioxygenase by fusing an endonuclease deactivated high-fidelity Cas9 (dHFCas9) to TET3 catalytic domain (TET3CD), targeted to specific genes by guiding RNAs (sgRNA). We demonstrate use of this technology in four different anti-fibrotic genes in different cell types in vitro, among them RASAL1 and Klotho, both hypermethylated in kidney fibrosis. Furthermore, in vivo lentiviral delivery of the Rasal1-targeted fusion protein to interstitial cells and of the Klotho-targeted fusion protein to tubular epithelial cells each results in specific gene reactivation and attenuation of fibrosis, providing gene-specific demethylating technology in a disease model

Serelaxin alleviates cardiac fibrosis through inhibiting endothelial-to-mesenchymal transition via RXFP1

Rationale: Cardiac fibrosis is an integral constituent of every form of chronic heart disease, and persistence of fibrosis reduces tissue compliance and accelerates the progression to heart failure. Relaxin-2 is a human hormone, which has various physiological functions such as mediating renal vasodilation in pregnancy. Its recombinant form Serelaxin has recently been tested in clinical trials as a therapy for acute heart failure but did not meet its primary endpoints. The aim of this study is to examine whether Serelaxin has an anti-fibrotic effect in the heart and therefore could be beneficial in chronic heart failure. Methods: We utilized two different cardiac fibrosis mouse models (ascending aortic constriction (AAC) and Angiotensin II (ATII) administration via osmotic minipumps) to assess the anti-fibrotic potential of Serelaxin. Histological analysis, immunofluorescence staining and molecular analysis were performed to assess the fibrosis level and indicate endothelial cells which are undergoing EndMT. In vitro TGF beta 1-induced endothelial-to-mesenchymal transition (EndMT) assays were performed in human coronary artery endothelial cells and mouse cardiac endothelial cells (MCECs) and were examined using molecular methods. Chromatin immunoprecipitation-qPCR assay was utilized to identify the Serelaxin effect on chromatin remodeling in the Rxfp1 promoter region in MCECs. Results: Our results demonstrate a significant and dose-dependent anti-fibrotic effect of Serelaxin in the heart in both models. We further show that Serelaxin mediates this effect, at least in part, through inhibition of EndMT through the endothelial Relaxin family peptide receptor 1 (RXFP1). We further demonstrate that Serelaxin administration is able to increase its own receptor expression (RXFP1) through epigenetic regulation in form of histone modifications by attenuating TGF beta-pSMAD2/3 signaling in endothelial cells. Conclusions: This study is the first to identify that Serelaxin increases the expression of its own receptor RXFP1 and that this mediates the inhibition of EndMT and cardiac fibrosis, suggesting that Serelaxin may have a beneficial effect as anti-fibrotic therapy in chronic heart failure

Probing the hadronic nature of the gamma-ray emission associated with Westerlund 2

Star-forming regions have been proposed as potential Galactic cosmic ray accelerators for decades. Cosmic ray acceleration can be probed through observations of gamma-rays produced in inelastic proton–proton collisions at GeV and TeV energies. In this paper, we analyse more than 11 yr of Fermi–LAT data from the direction of Westerlund 2, one of the most massive and best-studied star-forming regions in our Galaxy. In particular, we investigate the characteristics of the bright pulsar PSR J1023–5746 that dominates the gamma-ray emission below a few GeV at the position of Westerlund 2 and the underlying extended source FGES J1023.3–5747. The analysis results in a clear identification of FGES J1023.3–5747 as the GeV counterpart of the TeV source HESS J1023-575, through its morphological and spectral properties. This identification provides new clues about the origin of the HESS J1023-575 gamma-ray emission, favouring a hadronic origin of the emission, powered by Westerlund 2, rather than a leptonic origin related to either the pulsar wind nebula associated with PSR J1023–5746 or the cluster itself. This result indirectly supports the hypothesis that star-forming regions can contribute to the cosmic ray sea observed in our Galaxy

Probing the hadronic nature of the gamma-ray emission associated with Westerlund 2

Star-forming regions have been proposed as potential Galactic cosmic-ray accelerators for decades. Cosmic ray acceleration can be probed through observations of gamma-rays produced in inelastic proton-proton collisions, at GeV and TeV energies. We analyze more than 11 years of Fermi-LAT data from the direction of Westerlund 2, one of the most massive and best-studied star-forming regions in our Galaxy. The spectral and morphological characteristics of the LAT source agree with the ones in the TeV regime (HESS J1023-575), allowing the description of the gamma-ray source from a few hundreds of MeV to a few tens of TeVs. We will present the results and discuss the implications of the identification with the stellar cluster and the radiation mechanism involved

Revisiting the PeVatron candidate MGRO J1908+06 with an updated H.E.S.S. analysis

Detecting and studying galactic gamma-ray sources emitting very-high energy photons sheds light on the acceleration and propagation of cosmic rays presumably created in these sources. Currently, there are few sources emitting photons with energies exceeding 100 TeV. In this work we revisit the unidentified source MGRO J1908+06, initially detected by Milagro, using an updated H.E.S.S. dataset and analysis pipeline. The vicinity of the source contains a supernova remnant and pulsars as well as molecular clouds. This makes the identification of the primary source(s) of galactic cosmic rays as well as the nature of the gamma-ray emission challenging, especially in light of the recent HAWC and LHAASO detection of the high energy tail of its spectrum. Exploiting the better angular resolution as compared to particle detectors, we investigate the morphology of the source as well as its spectral properties

Constraints on the Intergalactic Magnetic Field Using Fermi-LAT and H.E.S.S. Blazar Observations

Magnetic fields in galaxies and galaxy clusters are believed to be the result of the amplification of intergalactic seed fields during the formation of large-scale structures in the universe. However, the origin, strength, and morphology of this intergalactic magnetic field (IGMF) remain unknown. Lower limits on (or indirect detection of) the IGMF can be obtained from observations of high-energy gamma rays from distant blazars. Gamma rays interact with the extragalactic background light to produce electron−positron pairs, which can subsequently initiate electromagnetic cascades. The gamma-ray signature of the cascade depends on the IGMF since it deflects the pairs. Here we report on a new search for this cascade emission using a combined data set from the Fermi Large Area Telescope and the High Energy Stereoscopic System. Using state-of-the-art Monte Carlo predictions for the cascade signal, our results place a lower limit on the IGMF of B > 7.1 × 10$^{−16}$ G for a coherence length of 1 Mpc even when blazar duty cycles as short as 10 yr are assumed. This improves on previous lower limits by a factor of 2. For longer duty cycles of 10$^{4}$ (10$^{7}$) yr, IGMF strengths below 1.8 × 10$^{−14}$ G (3.9 × 10$^{−14}$ G) are excluded, which rules out specific models for IGMF generation in the early universe

Revisiting the PeVatron candidate MGRO J1908+06 with an updated H.E.S.S. analysis

Detecting and studying galactic gamma-ray sources emitting very-high energy photons sheds light on the acceleration and propagation of cosmic rays presumably created in these sources. Currently, there are few sources emitting photons with energies exceeding 100 TeV. In this work we revisit the unidentified source MGRO J1908+06, initially detected by Milagro, using an updated H.E.S.S. dataset and analysis pipeline. The vicinity of the source contains a supernova remnant and pulsars as well as molecular clouds. This makes the identification of the primary source(s) of galactic cosmic rays as well as the nature of the gamma-ray emission challenging, especially in light of the recent HAWC and LHAASO detection of the high energy tail of its spectrum. Exploiting the better angular resolution as compared to particle detectors, we investigate the morphology of the source as well as its spectral properties

H.E.S.S. Follow-up Observations of Binary Black Hole Coalescence Events during the Second and Third Gravitational-wave Observing Runs of Advanced LIGO and Advanced Virgo

We report on the observations of four well-localized binary black hole (BBH) mergers by the High Energy Stereoscopic System (H.E.S.S.) during the second and third observing runs of Advanced LIGO and Advanced Virgo, O2 and O3. H.E.S.S. can observe 20 deg$^{2}$ of the sky at a time and follows up gravitational-wave (GW) events by “tiling” localization regions to maximize the covered localization probability. During O2 and O3, H.E.S.S. observed large portions of the localization regions, between 35% and 75%, for four BBH mergers (GW170814, GW190512_180714, GW190728_064510, and S200224ca). For these four GW events, we find no significant signal from a pointlike source in any of the observations, and we set upper limits on the very high energy (>100 GeV) γ-ray emission. The 1–10 TeV isotropic luminosity of these GW events is below 10$^{45}$ erg s$^{−1}$ at the times of the H.E.S.S. observations, around the level of the low-luminosity GRB 190829A. Assuming no changes are made to how follow-up observations are conducted, H.E.S.S. can expect to observe over 60 GW events per year in the fourth GW observing run, O4, of which eight would be observable with minimal latency