Zika Virus Disrupts Molecular Fingerprinting Of Human Neurospheres

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Zika virus (ZIKV) has been associated with microcephaly and other brain abnormalities; however, the molecular consequences of ZIKV to human brain development are still not fully understood. Here we describe alterations in human neurospheres derived from induced pluripotent stem (iPS) cells infected with the strain of Zika virus that is circulating in Brazil. Combining proteomics and mRNA transcriptional profiling, over 500 proteins and genes associated with the Brazilian ZIKV infection were found to be differentially expressed. These genes and proteins provide an interactome map, which indicates that ZIKV controls the expression of RNA processing bodies, miRNA biogenesis and splicing factors required for self-replication. It also suggests that impairments in the molecular pathways underpinning cell cycle and neuronal differentiation are caused by ZIKV. These results point to biological mechanisms implicated in brain malformations, which are important to further the understanding of ZIKV infection and can be exploited as therapeutic potential targets to mitigate it.7Brazilian Development Bank (BNDES)Funding Authority for Studies and Projects (FINEP)National Council of Scientific and Technological Development (CNPq)Foundation for Research Support in the State of Rio de Janeiro (FAPERJ)Sao Paulo Research Foundation (FAPESP) [14/21035-0, 14/14881-1, 13/08711-3, 14/10068-4]Coordination for the Improvement of Higher Education Personnel (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

The cyanobacterial saxitoxin exacerbates neural cell death and brain malformations induced by zika virus

The northeast (NE) region of Brazil commonly goes through drought periods, which favor cyanobacterial blooms, capable of producing neurotoxins with implications for human and animal health. The most severe dry spell in the history of Brazil occurred between 2012 and 2016. Coincidently, the highest incidence of microcephaly associated with the Zika virus (ZIKV) outbreak took place in the NE region of Brazil during the same years. In this work, we tested the hypothesis that saxitoxin (STX), a neurotoxin produced in South America by the freshwater cyanobacteria Raphidiopsis raciborskii, could have contributed to the most severe Congenital Zika Syndrome (CZS) profile described worldwide. Quality surveillance showed higher cyanobacteria amounts and STX occurrence in human drinking water sup-plies of NE compared to other regions of Brazil. Experimentally, we described that STX dou-bled the quantity of ZIKV-induced neural cell death in progenitor areas of human brain organoids, while the chronic ingestion of water contaminated with STX before and during gestation caused brain abnormalities in offspring of ZIKV-infected immunocompetent C57BL/6J mice. Our data indicate that saxitoxin-producing cyanobacteria is overspread in water reservoirs of the NE and might have acted as a co-insult to ZIKV infection in Brazil. These results raise a public health concern regarding the consequences of arbovirus outbreaks happening in areas with droughts and/or frequent freshwater cyanobacterial blooms.Fil: Pedrosa, Carolina da S. G.. D’Or Institute for Research and Education; BrasilFil: Souza, Leticia R. Q.. D’Or Institute for Research and Education; BrasilFil: Gomes, Tiago A.. Universidade Federal do Rio de Janeiro; Brasil. Instituto Oswaldo Cruz; BrasilFil: de Lima, Caroline V. F.. D’Or Institute for Research and Education; BrasilFil: Ledur, Pitia F.. D’Or Institute for Research and Education; BrasilFil: Karmirian, Karina. D’Or Institute for Research and Education; Brasil. Universidade Federal do Rio de Janeiro; BrasilFil: Barbeito Andrés, Jimena. Universidade Federal do Rio de Janeiro; Brasil. Universidad Nacional Arturo Jauretche. Unidad Ejecutora de Estudios en Neurociencias y Sistemas Complejos. Provincia de Buenos Aires. Ministerio de Salud. Hospital Alta Complejidad en Red El Cruce Dr. Néstor Carlos Kirchner Samic. Unidad Ejecutora de Estudios en Neurociencias y Sistemas Complejos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Unidad Ejecutora de Estudios en Neurociencias y Sistemas Complejos; ArgentinaFil: Costa, Marcelo do N.. Universidade Federal do Rio de Janeiro; BrasilFil: Higa, Luiza M.. Universidade Federal do Rio de Janeiro; BrasilFil: Rossi, Átila D.. Universidade Federal do Rio de Janeiro; BrasilFil: Bellio, Maria. Universidade Federal do Rio de Janeiro; BrasilFil: Tanuri, Amilcar. Universidade Federal do Rio de Janeiro; BrasilFil: Prata Barbosa, Arnaldo. D’Or Institute for Research and Education; BrasilFil: Tovar Moll, Fernanda. D’Or Institute for Research and Education; Brasil. Universidade Federal do Rio de Janeiro; BrasilFil: Garcez, Patricia P.. Universidade Federal do Rio de Janeiro; BrasilFil: Lara, Flavio A.. Instituto Oswaldo Cruz; BrasilFil: Molica, Renato J. R.. Universidad Federal Rural Pernambuco; BrasilFil: Rehen, Stevens K.. D’Or Institute for Research and Education; Brasil. Universidade Federal do Rio de Janeiro; Brasi

Rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART): Study protocol for a randomized controlled trial

Background: Acute respiratory distress syndrome (ARDS) is associated with high in-hospital mortality. Alveolar recruitment followed by ventilation at optimal titrated PEEP may reduce ventilator-induced lung injury and improve oxygenation in patients with ARDS, but the effects on mortality and other clinical outcomes remain unknown. This article reports the rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART). Methods/Design: ART is a pragmatic, multicenter, randomized (concealed), controlled trial, which aims to determine if maximum stepwise alveolar recruitment associated with PEEP titration is able to increase 28-day survival in patients with ARDS compared to conventional treatment (ARDSNet strategy). We will enroll adult patients with ARDS of less than 72 h duration. The intervention group will receive an alveolar recruitment maneuver, with stepwise increases of PEEP achieving 45 cmH(2)O and peak pressure of 60 cmH2O, followed by ventilation with optimal PEEP titrated according to the static compliance of the respiratory system. In the control group, mechanical ventilation will follow a conventional protocol (ARDSNet). In both groups, we will use controlled volume mode with low tidal volumes (4 to 6 mL/kg of predicted body weight) and targeting plateau pressure <= 30 cmH2O. The primary outcome is 28-day survival, and the secondary outcomes are: length of ICU stay; length of hospital stay; pneumothorax requiring chest tube during first 7 days; barotrauma during first 7 days; mechanical ventilation-free days from days 1 to 28; ICU, in-hospital, and 6-month survival. ART is an event-guided trial planned to last until 520 events (deaths within 28 days) are observed. These events allow detection of a hazard ratio of 0.75, with 90% power and two-tailed type I error of 5%. All analysis will follow the intention-to-treat principle. Discussion: If the ART strategy with maximum recruitment and PEEP titration improves 28-day survival, this will represent a notable advance to the care of ARDS patients. Conversely, if the ART strategy is similar or inferior to the current evidence-based strategy (ARDSNet), this should also change current practice as many institutions routinely employ recruitment maneuvers and set PEEP levels according to some titration method.Hospital do Coracao (HCor) as part of the Program 'Hospitais de Excelencia a Servico do SUS (PROADI-SUS)'Brazilian Ministry of Healt

Cenpj/CPAP regulates progenitor divisions and neuronal migration in the cerebral cortex downstream of Ascl1

The proneural factor Ascl1 controls multiple steps of neurogenesis in the embryonic brain, including progenitor division and neuronal migration. Here we show that Cenpj, also known as CPAP, a microcephaly gene, is a transcriptional target of Ascl1 in the embryonic cerebral cortex. We have characterized the role of Cenpj during cortical development by in utero electroporation knockdown and found that silencing Cenpj in the ventricular zone disrupts centrosome biogenesis and randomizes the cleavage plane orientation of radial glia progenitors. Moreover, we show that downregulation of Cenpj in post-mitotic neurons increases stable microtubules and leads to slower neuronal migration, abnormal centrosome position and aberrant neuronal morphology. Moreover, rescue experiments shows that Cenpj mediates the role of Ascl1 in centrosome biogenesis in progenitor cells and in microtubule dynamics in migrating neurons. These data provide insights into genetic pathways controlling cortical development and primary microcephaly observed in humans with mutations in Cenpj

Loss of cannabinoid CB1 receptors induces cortical migration malformations and increases seizure susceptibility

Neuronal migration is a fundamental process of brain development, and its disruption underlies devastating neurodevelopmental disorders. The transcriptional programs governing this process are relatively well characterized. However, how environmental cues instruct neuronal migration remains poorly understood. Here, we demonstrate that the cannabinoid CB1 receptor is strictly required for appropriate pyramidal neuron migration in the developing cortex. Acute silencing of the CB1 receptor alters neuronal morphology and impairs radial migration. Consequently, CB1 siRNAelectroporated mice display cortical malformations mimicking subcortical band heterotopias and increased seizure susceptibility in adulthood. Importantly, rescuing the CB1 deficiency-induced radial migration arrest by knockdown of the GTPase protein RhoA restored the hyperexcitable neuronal network and seizure susceptibility. Our findings show that CB1 receptor/RhoA signaling regulates pyramidal neuron migration, and that deficient CB1 receptor signaling may contribute to cortical development malformations leading to refractory epilepsy independently of its canonical neuromodulatory role in the adult brain

TITLE: Zika virus impairs growth in human neurospheres and brain organoids

SUMMARY: We provide evidence that Zika virus infects human iPS-derived neural stem cells, causing cell death and reduced growth in neurospheres and cerebral organoids

Zika virus disrupts molecular fingerprinting of human neurospheres

Zika virus (ZIKV) has been associated with microcephaly and other brain abnormalities; however, the molecular consequences of ZIKV to human brain development are still not fully understood. Here we describe alterations in human neurospheres derived from induced pluripotent stem (iPS) cells infected with the strain of Zika virus that is circulating in Brazil. Combining proteomics and mRNA transcriptional profiling, over 500 proteins and genes associated with the Brazilian ZIKV infection were found to be differentially expressed. These genes and proteins provide an interactome map, which indicates that ZIKV controls the expression of RNA processing bodies, miRNA biogenesis and splicing factors required for self-replication. It also suggests that impairments in the molecular pathways underpinning cell cycle and neuronal differentiation are caused by ZIKV. These results point to biological mechanisms implicated in brain malformations, which are important to further the understanding of ZIKV infection and can be exploited as therapeutic potential targets to mitigate it7CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFINANCIADORA DE ESTUDOS E PROJETOS - FINEPFUNDAÇÃO CARLOS CHAGAS FILHO DE AMPARO À PESQUISA DO ESTADO DO RIO DE JANEIRO - FAPERJFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informaçãosem informaçãosem informaçãosem informação14/21035-0; 14/14881-1; 13/08711-3; 14/10068-

Zika virus disrupts molecular fingerprinting of human neurospheres

Submitted by Sandra Infurna ([email protected]) on 2018-04-03T16:26:16Z No. of bitstreams: 1 anamariab_filippis_etal_IOC_2017.pdf: 1347352 bytes, checksum: b3a68b2d8ea28413682153a717faa4c4 (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2018-04-03T16:44:04Z (GMT) No. of bitstreams: 1 anamariab_filippis_etal_IOC_2017.pdf: 1347352 bytes, checksum: b3a68b2d8ea28413682153a717faa4c4 (MD5)Made available in DSpace on 2018-04-03T16:44:04Z (GMT). No. of bitstreams: 1 anamariab_filippis_etal_IOC_2017.pdf: 1347352 bytes, checksum: b3a68b2d8ea28413682153a717faa4c4 (MD5) Previous issue date: 2017Instituto D´Or de Pesquisa e Educação. Rio de Janeiro, Brasil / Universidade Federal do Rio de Janeiro. Instituto de Ciências Biológicas. Rio de Janeiro, RJ, Brasil.Instituto D´Or de Pesquisa e Educação. Rio de Janeiro, Brasil / Universidade de Campinas. Departamento de Bioquímica e Biologia Tecidual. Campinas, SP, Brasil.Instituto Evandro Chagas. Centro de Inovação Tecnológica. Belém, PA, BrasilInstituto D´Or de Pesquisa e Educação. Rio de Janeiro, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Biologia. Rio de Janeiro, RJ, BrasilInstituto D´Or de Pesquisa e Educação. Rio de Janeiro, Brasil.Instituto D´Or de Pesquisa e Educação. Rio de Janeiro, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Biologia. Rio de Janeiro, RJ, BrasilUniversidade de Campinas. Departamento de Bioquímica e Biologia Tecidual. Campinas, SP, Brasil.Instituto D´Or de Pesquisa e Educação. Rio de Janeiro, Brasil.Fundação Oswaldo Cruz Fiocruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Instituto D´Or de Pesquisa e Educação. Rio de Janeiro, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Biologia. Rio de Janeiro, RJ, BrasilUniversidade Federal do Pará. Belém, PA, Brasil.Fundação Oswaldo Cruz Fiocruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Instituto Evandro Chagas. Centro de Inovação Tecnológica. Belém, PA, BrasilUniversidade Federal do Rio de Janeiro. Instituto de Biologia. Rio de Janeiro, RJ, BrasilUniversidade de Campinas. Departamento de Bioquímica e Biologia Tecidual. Campinas, SP, Brasil.Instituto D´Or de Pesquisa e Educação. Rio de Janeiro, Brasil / Universidade Federal do Rio de Janeiro. Instituto de Ciências Biológicas. Rio de Janeiro, RJ, Brasil.Zika virus (ZIKV) has been associated with microcephaly and other brain abnormalities; however, the molecular consequences of ZIKV to human brain development are still not fully understood. Here we describe alterations in human neurospheres derived from induced pluripotent stem (iPS) cells infected with the strain of Zika virus that is circulating in Brazil. Combining proteomics and mRNA transcriptional profiling, over 500 proteins and genes associated with the Brazilian ZIKV infection were found to be differentially expressed. These genes and proteins provide an interactome map, which indicates that ZIKV controls the expression of RNA processing bodies, miRNA biogenesis and splicing factors required for self-replication. It also suggests that impairments in the molecular pathways underpinning cell cycle and neuronal differentiation are caused by ZIKV. These results point to biological mechanisms implicated in brain malformations, which are important to further the understanding of ZIKV infection and can be exploited as therapeutic potential targets to mitigate it

Congenital Zika syndrome is associated with maternal protein malnutrition

MRC Zika Rapid Response Grant (MC_PC_15102), FAPERJ (E_03/2017E_03/2017), the Brazilian Ministry of Health (DSAST/SVS 25380.001612/2017-70), and the AAPA Professional Development GrantFederal University of Rio de Janeiro. Institute of Biomedical Sciences. Rio de Janeiro, RJ, Brazil / Institute for Studies in Neuroscience and Complex Systems. Buenos Aires, Argentina.Federal University of Rio de Janeiro. Institute of Biology. Department of Genetics. Rio de Janeiro, RJ, Brazil.Federal University of Rio de Janeiro. Institute of Biology. Department of Genetics. Rio de Janeiro, RJ, Brazil.Federal University of Rio de Janeiro. Microbiology Institute Paulo de Góes. Rio de Janeiro, RJ, Brazil.Federal University of Pará. Biological Science Institute. Belém, PA, Brazil.Federal University of Rio de Janeiro. Nuclear Instrumentation Laboratory. Rio de Janeiro RJ, Brazil.Federal University of Rio de Janeiro. Institute of Biomedical Sciences. Rio de Janeiro, RJ, Brazil.Federal University of Rio de Janeiro. Institute of Biomedical Sciences. Rio de Janeiro, RJ, Brazil.Federal University of Rio de Janeiro. Microbiology Institute Paulo de Góes. Rio de Janeiro, RJ, Brazil.Federal University of Rio de Janeiro. Institute of Biology. Department of Genetics. Rio de Janeiro, RJ, Brazil.National Center of Structural Biology and Bioimaging. Rio de Janeiro, RJ, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações tecnológicas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações tecnológicas. Ananindeua, PA, Brasil.State Institute of Brain Paulo Niemeyer. Rio de Janeiro, RJ, Brazil.Federal University of Rio de Janeiro. Institute of Biology. Department of Genetics. Rio de Janeiro, RJ, Brazil.Institute for Studies in Neuroscience and Complex Systems. Buenos Aires, Argentina.Fundação Oswaldo Cruz. Oswaldo Cruz Institute. Rio de Janeiro, RJ, Brazil.Harvard T. H. Chan School of Public Health. Department of Global Health and Population. Boston, MA, USA.Zika virus (ZIKV) infection during pregnancy is associated with a spectrum of developmental impairments known as congenital Zika syndrome (CZS). The prevalence of this syndrome varies across ZIKV endemic regions, suggesting that its occurrence could depend on cofactors. Here, we evaluate the relevance of protein malnutrition for the emergence of CZS. Epidemiological data from the ZIKV outbreak in the Americas suggest a relationship between undernutrition and cases of microcephaly. To experimentally examine this relationship, we use immunocompetent pregnant mice, which were subjected to protein malnutrition and infected with a Brazilian ZIKV strain. We found that the combination of protein restriction and ZIKV infection leads to severe alterations of placental structure and embryonic body growth, with offspring displaying a reduction in neurogenesis and postnatal brain size. RNA-seq analysis reveals gene expression deregulation required for brain development in infected low-protein progeny. These results suggest that maternal protein malnutrition increases susceptibility to CZS