Epigenetic regulation of nitric oxide synthase 2, inducible (Nos2) by NLRC4 inflammasomes involves PARP1 cleavage

Nitric oxide synthase 2, inducible (Nos2) expression is necessary for the microbicidal activity of macrophages. However, NOS2 over-activation causes multiple inflammatory disorders, suggesting a tight gene regulation is necessary. Using cytosolic flagellin as a model for inflammasome-dependent NOS2 activation, we discovered a surprising new role for NLRC4/caspase-1 axis in regulating chromatin accessibility of the Nos2 promoter. We found that activation of two independent mechanisms is necessary for NOS2 expression by cytosolic flagellin: caspase-1 and NF-kappa B activation. NF-kappa B activation was necessary, but not sufficient, for NOS2 expression. Conversely, caspase-1 was necessary for NOS2 expression, but dispensable for NF-kappa B activation, indicating that this protease acts downstream NF-kappa B activation. We demonstrated that epigenetic regulation of Nos2 by caspase-1 involves cleavage of the chromatin regulator PARP1 (also known as ARTD1) and chromatin accessibility of the NF-kappa B binding sites located at the Nos2 promoter. Remarkably, caspase-1-mediated Nos2 transcription and NO production contribute to the resistance of macrophages to Salmonella typhimurium infection. Our results uncover the molecular mechanism behind the constricted regulation of Nos2 expression and open new therapeutic opportunities based on epigenetic activities of caspase-1 against infectious and inflammatory diseases.Kanton of ZurichUniversity Research Priority Program (URPP) in Translational Cancer Biology at the University of ZurichSwiss National Science FoundationCancer Research SocietyCanadian Cancer SocietyNSERCOntario Institute for Cancer Research (OICR)province of OntarioPrincess Margaret Cancer FoundationUniversity of Toronto McLaughlin CentreFundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP - Brazil)Brazilian Research Council (CNPq-Brazil)CAPESINCTVUniv Fed Sao Paulo, Ctr Terapia Celular & Mol CTC Mol, Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Ciencias Biol, Sao Paulo, BrazilUniv Sao Paulo, Inst Ciencias Biomed, Dept Microbiol, Sao Paulo, BrazilUniv Hlth Network, Princess Margaret Canc Ctr, Toronto, ON M5G 2M9, CanadaUniv Sao Paulo, Inst Ciencias Biomed, Sao Paulo & Inst Invest Imunol, Inst Nacl Ciencia Tecnol INCT 3, Sao Paulo, BrazilInst Nacl Ciencia Tecnol INCT III, Inst Invest Imunol, Sao Paulo, BrazilUniv Zurich, Dept Mol Mech Dis, Zurich, SwitzerlandUniv Toronto, Dept Med Biophys, Toronto, ON M5G 2M9, CanadaUniv Fed Sao Paulo, Ctr Terapia Celular & Mol CTC Mol, Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Ciencias Biol, Sao Paulo, BrazilSwiss National Science Foundation: 310030B_138667Cancer Research Society: CRS19092Cancer Research Society: CRS19091Canadian Cancer Society: CCSRI 703279Canadian Cancer Society CCSRI 703716NSERC: 489073University of Toronto McLaughlin Centre: MC-2015-02FAPESP: 2013/16010-5FAPESP: 2015/18003-1Web of Scienc

Evaluation of the molecular mechanisms involved in the iNOS expression by NAIP5/NLRC4-Caspase-1 axis.

O reconhecimento da flagelina é compartilhado pelo receptor transmembrânico TLR5 e citosólico NAIP5/NLRC4. Entretanto, pouco se sabe sobre os mecanismos efetores individuais induzidos a partir do reconhecimento extra e intracelular da flagelina. Aqui, nós demonstramos que macrófagos estimulados com a flagelina citosólica (FLA-BSDot) induziu a expressão de iNOS, enzima responsável pela produção do óxido nítrico (NO). A expressão de iNOS foi dependente do eixo NAIP5/NLRC4/caspase-1 e independente de IL-1β, IL-18 e MyD88, descartando a via de ativação dos TLRs. Ainda, esta via não requer a ativação do fator de transcrição IRF-1, mas envolve a ativação do NF-kB, assim como a clivagem da enzima PARP-1 (poly(ADP-ribose)polymerase-1). Por fim, avaliamos a relevância biológica desta via no controle das infecções por L. pneumophila e S. Typhimurium, dados que definem um mecanismo efetor adicional no controle de patógenos.Recognition of flagellin is shared by transmembranic TLR5 and cytosolic NAIP5/NLRC4. However, little is known about the individual effector mechanisms induced by extra and intracellular flagellin. Here, we have demonstrated that cytosolic flagellin-stimulated macrophages (FLA-BSDot) induced iNOS expression, an enzyme responsible for the production of nitric oxide (NO). iNOS expression was dependent of the NAIP5/NLRC4/caspase-1 axis and independent of IL-1β, IL-18 and MyD88, discarding TLRs signaling pathway. Still, this pathway do not require the activation of IRF-1 transcriptional factor, but involves NF-kB activation as well as the cleavage of the enzyme, PARP-1 (poly(ADP-ribose)polymerase-1). Finally, we have evaluated the biological relevance of this pathway in the control of the infections by L. pneumophila e S. Typhimurium, which define an additional effector mechanism to the control of pathogens

Cytosolic flagellin-induced lysosomal pathway regulates inflammasome-dependent and -independent macrophage responses

NAIP5/NLRC4 (neuronal apoptosis inhibitory protein 5/nucleotide oligomerization domain-like receptor family, caspase activation recruitment domain domain-containing 4) inflammasome activation by cytosolic flagellin results in caspase-1-mediated processing and secretion of IL-1β/IL-18 and pyroptosis, an inflammatory cell death pathway. Here, we found that although NLRC4, ASC, and caspase-1 are required for IL-1β secretion in response to cytosolic flagellin, cell death, nevertheless, occurs in the absence of these molecules. Cytosolic flagellin-induced inflammasome-independent cell death is accompanied by IL-1α secretion and is temporally correlated with the restriction of Salmonella Typhimurium infection. Despite displaying some apoptotic features, this peculiar form of cell death do not require caspase activation but is regulated by a lysosomal pathway, in which cathepsin B and cathepsin D play redundant roles. Moreover, cathepsin B contributes to NAIP5/NLRC4 inflammasome-induced pyroptosis and IL-1α and IL-1β production in response to cytosolic flagellin. Together, our data describe a pathway induced by cytosolic flagellin that induces a peculiar form of cell death and regulates inflammasome-mediated effector mechanisms of macrophagesFundação de Amparo à Pesquisa do Estado de São PauloBrazilian Research Counci

Cellular Renewal and Improvement of Local Cell Effector Activity in Peritoneal Cavity in Response to Infectious Stimuli

The peritoneal cavity (PerC) is a singular compartment where many cell populations reside and interact. Despite the widely adopted experimental approach of intraperitoneal (i.p.) inoculation, little is known about the behavior of the different cell populations within the PerC. To evaluate the dynamics of peritoneal macrophage (Mempty set) subsets, namely small peritoneal Mempty set (SPM) and large peritoneal Mempty set (LPM), in response to infectious stimuli, C57BL/6 mice were injected i.p. with zymosan or Trypanosoma cruzi. These conditions resulted in the marked modification of the PerC myelo-monocytic compartment characterized by the disappearance of LPM and the accumulation of SPM and monocytes. In parallel, adherent cells isolated from stimulated PerC displayed reduced staining for beta-galactosidase, a biomarker for senescence. Further, the adherent cells showed increased nitric oxide (NO) and higher frequency of IL-12-producing cells in response to subsequent LPS and IFN-gamma stimulation. Among myelo-monocytic cells, SPM rather than LPM or monocytes, appear to be the central effectors of the activated PerC; they display higher phagocytic activity and are the main source of IL-12. Thus, our data provide a first demonstration of the consequences of the dynamics between peritoneal Mempty set subpopulations by showing that substitution of LPM by a robust SPM and monocytes in response to infectious stimuli greatly improves PerC effector activity.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP - Brazil)[08/50958-8]Brazilian Research Council (CNPq-Brazil)Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES

The Southern Photometric Local Universe Survey (S-PLUS): improved SEDs, morphologies, and redshifts with 12 optical filters

The Southern Photometric Local Universe Survey (S-PLUS) is imaging similar to 9300 deg(2) of the celestial sphere in 12 optical bands using a dedicated 0.8mrobotic telescope, the T80-South, at the Cerro Tololo Inter-american Observatory, Chile. The telescope is equipped with a 9.2k x 9.2k e2v detector with 10 mu m pixels, resulting in a field of view of 2 deg(2) with a plate scale of 0.55 arcsec pixel-1. The survey consists of four main subfields, which include two non-contiguous fields at high Galactic latitudes (vertical bar b vertical bar > 30 degrees, 8000 deg(2)) and two areas of the Galactic Disc and Bulge (for an additional 1300 deg(2)). S-PLUS uses the Javalambre 12-band magnitude system, which includes the 5 ugriz broad-band filters and 7 narrow-band filters centred on prominent stellar spectral features: the Balmer jump/[OII], Ca H + K, Hd, G band, Mg b triplet, H alpha, and the Ca triplet. S-PLUS delivers accurate photometric redshifts (dz /(1 + z) = 0.02 or better) for galaxies with r < 19.7 AB mag and z < 0.4, thus producing a 3D map of the local Universe over a volume of more than 1 (Gpc/h)(3). The final S-PLUS catalogue will also enable the study of star formation and stellar populations in and around the Milky Way and nearby galaxies, as well as searches for quasars, variable sources, and low-metallicity stars. In this paper we introduce the main characteristics of the survey, illustrated with science verification data highlighting the unique capabilities of S-PLUS. We also present the first public data release of similar to 336 deg(2) of the Stripe 82 area, in 12 bands, to a limiting magnitude of r = 21, available at datalab.noao.edu/splus.© 2019 The Author(s).Published by Oxford University Press on behalf of the Royal Astronomical SocietyThe S-PLUS project, including the T80S robotic telescope and the S-PLUS scientific survey, was founded as a partnership between the Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP), the Observatorio Nacional (ON), the Federal University of Sergipe (UFS), and the Federal University of Santa Catarina (UFSC), with important financial and practical contributions from other collaborating institutes in Brazil, Chile (Universidad de La Serena), and Spain (Centro de Estudios de Fisica del Cosmos de Aragon, CEFCA). The members of the collaboration are grateful for the support received from the Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq; grants 312333/2014-5, 306968/2014-2, 142436/2014-3, 459553/2014-3, 400738/2014-7, 302037/2015-2, 312307/2015-2, 300336/2016-0, 304184/2016-0, 304971/2016-2, 401669/2016-5, 308968/2016-6, 309456/2016-9, 421687/2016-9, 150237/2017-0, 311331/2017-3, 304819/2017-4, and 200289/2017-9), FAPESP (grants 2009/54202-8, 2011/51680-6, 2014/07684-5, 2014/11806-9, 2014/13723-3, 2014/18632-6, 2016/17119-9, 2016/12331-0, 2016/21532-9, 2016/21664-2, 2016/23567-4, 2017/01461-2, 2017/23766-0, 2018/02444-7, and 2018/21661-9), the Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES; grants 88881.030413/2013-01 and 88881.156185/2017-01), the Fundacao de Amparo a Pesquisa do Estado do Rio de Janeiro (FAPERJ; grants 202.876/2015, 202.835/2016, and 203.186/2016), the Financiadora de Estudos e Projetos (FINEP; grants 1217/13-01.13.0279.00 and 0859/10-01.10.0663.00), the Direccion de Investigacion y Desarrollo de la Universidad de La Serena (DIDULS/ULS; projects PR16143 and PTE16146 and the Programa de Investigadores Asociados), and the Direccion de Postgrado y Postitulo. TCB, VMP, and DDW acknowledge the support from the Physics Frontier Center for the Evolution of the Elements (JINA-CEE) through the US National Science Foundation (grant PHY 14-30152). JLNC is grateful for financial support received from the Southern Office of Aerospace Research and development (SOARD; grants FA9550-15-1-0167 and FA9550-18-1-0018) of the Air Force Office of the Scientific Research International Office of the United States (AFOSR/IO). YJT and RAD acknowledge support from the Spanish National Research Council (CSIC) I-COOP + 2016 program (grant COOPB20263), and the Spanish Ministry of Economy, Industry, and Competitiveness (MINECO; grants AYA2013-48623-C2-1-P and AYA2016-81065-C2-1-P). RAOM acknowledges support from the Direccion General de Asuntos del Personal Academico of the Universidad Nacional Autonoma de Mexico (DGAPA-UNAM) through a post-doctoral fellowship from the Programa de Becas Posdoctorales en la UNAM. This work has made use of data from the Sloan Digital Sky Survey. Funding for the SDSS and SDSS-II has been provided by the Alfred P. Sloan Foundation, the Participating Institutions, the National Science Foundation, the U.S. Department of Enenergy, the National Aeronautics and Space Administration, the Japanese Monbukagakusho, the Max Planck Society, and the Higher Education Funding Council for England. The SDSS Web Site is http://www.sdss.org/.The SDSS is managed by the Astrophysical Research Consortium for the Participating Institutions. The Participating Institutions are the American Museum of Natural History, Astrophysical Institute Potsdam, University of Basel, University of Cambridge, Case Western Reserve University, University of Chicago, Drexel University, Fermilab, the Institute for Advanced Study, the Japan Participation Group, Johns Hopkins University, the Joint Institute for Nuclear Astrophysics, the Kavli Institute for Particle Astrophysics and Cosmology, the Korean Scientist Group, the Chinese Academy of Sciences (LAMOST), Los Alamos National Laboratory, the Max-Planck-Institute for Astronomy (MPIA), the Max-Planck-Institute for Astrophysics (MPA), the New Mexico State University, Ohio State University, University of Pittsburgh, University of Portsmouth, Princeton University, the United States Naval Observatory, and the University of Washington. This publication makes use of data products from the Widefield Infrared Survey Explorer, which is a joint project of the University of California, Los Angeles, and the Jet Propulsion Laboratory/California Institute of Technology, funded by the National Aeronautics and Space Administration. We are grateful for the contributions of CTIO staff in helping in the construction, commissioning, and maintenance of the telescope and camera and we are particularly thankful to the CTIO director, Steve Heathcote, for his support at every phase, without which this project would not have been completed. We thank Cesar Iniguez for making the 2D measurements of the filter transmissions at CEFCA. We warmly thank David Cristobal-Hornillos and his group for helping us to install and run the reduction package JYPE version 0.9.9 in the S-PLUS computer system in Chile. We warmly thank Mariano Moles, Javier Cenarro, Tamara Civera, Sergio Chueca, Javier Hernandez Fuertes, Antonio Marin Franch, Jesus Varella, and Hector Vazquez Ramio -the success of the S-PLUS project relies on the dedication of these and other CEFCA staff members in building OAJ and running J-PLUS and J-PAS. We deeply thank Rene Laporte and INPE, as well as Keith Taylor, for their contributions to the T80S camera