TY - JOUR
AB - The complexity of the microenvironment effects on cell response, show accumulating evidence that glioblastoma (GBM) migration and invasiveness are influenced by the mechanical rigidity of their surroundings. The epithelial–mesenchymal transition (EMT) is a well-recognized driving force of the invasive behavior of cancer. However, the primary mechanisms of EMT initiation and progression remain unclear. We have previously showed that certain substrate stiffness can selectively stimulate human GBM U251-MG and GL15 glioblastoma cell lines motility. The present study unifies several known EMT mediators to uncover the reason of the regulation and response to these stiffnesses. Our results revealed that changing the rigidity of the mechanical environment tuned the response of both cell lines through change in morphological features, epithelial-mesenchymal markers (E-, N-Cadherin), EGFR and ROS expressions in an interrelated manner. Specifically, a stiffer microenvironment induced a mesenchymal cell shape, a more fragmented morphology, higher intracellular cytosolic ROS expression and lower mitochondrial ROS. Finally, we observed that cells more motile showed a more depolarized mitochondrial membrane potential. Unravelling the process that regulates GBM cells’ infiltrative behavior could provide new opportunities for identification of new targets and less invasive approaches for treatment.
AU - Basilico, Bernadette
AU - Palamà, Ilaria Elena
AU - D’Amone, Stefania
AU - Lauro, Clotilde
AU - Rosito, Maria
AU - Grieco, Maddalena
AU - Ratano, Patrizia
AU - Cordella, Federica
AU - Sanchini, Caterina
AU - Di Angelantonio, Silvia
AU - Ragozzino, Davide
AU - Cascione, Mariafrancesca
AU - Gigli, Giuseppe
AU - Cortese, Barbara
ID - 12268
JF - Frontiers in Oncology
KW - Cancer Research
KW - Oncology
SN - 2234-943X
TI - Substrate stiffness effect on molecular crosstalk of epithelial-mesenchymal transition mediators of human glioblastoma cells
VL - 12
ER -
TY - JOUR
AB - Microglia cells are active players in regulating synaptic development and plasticity in the brain. However, how they influence the normal functioning of synapses is largely unknown. In this study, we characterized the effects of pharmacological microglia depletion, achieved by administration of PLX5622, on hippocampal CA3-CA1 synapses of adult wild type mice. Following microglial depletion, we observed a reduction of spontaneous and evoked glutamatergic activity associated with a decrease of dendritic spine density. We also observed the appearance of immature synaptic features and higher levels of plasticity. Microglia depleted mice showed a deficit in the acquisition of the Novel Object Recognition task. These events were accompanied by hippocampal astrogliosis, although in the absence ofneuroinflammatory condition. PLX-induced synaptic changes were absent in Cx3cr1−/− mice, highlighting the role of CX3CL1/CX3CR1 axis in microglia control of synaptic functioning. Remarkably, microglia repopulation after PLX5622 withdrawal was associated with the recovery of hippocampal synapses and learning functions. Altogether, these data demonstrate that microglia contribute to normal synaptic functioning in the adult brain and that their removal induces reversible changes in organization and activity of glutamatergic synapses.
AU - Basilico, Bernadette
AU - Ferrucci, Laura
AU - Ratano, Patrizia
AU - Golia, Maria T.
AU - Grimaldi, Alfonso
AU - Rosito, Maria
AU - Ferretti, Valentina
AU - Reverte, Ingrid
AU - Sanchini, Caterina
AU - Marrone, Maria C.
AU - Giubettini, Maria
AU - De Turris, Valeria
AU - Salerno, Debora
AU - Garofalo, Stefano
AU - St‐Pierre, Marie‐Kim
AU - Carrier, Micael
AU - Renzi, Massimiliano
AU - Pagani, Francesca
AU - Modi, Brijesh
AU - Raspa, Marcello
AU - Scavizzi, Ferdinando
AU - Gross, Cornelius T.
AU - Marinelli, Silvia
AU - Tremblay, Marie‐Ève
AU - Caprioli, Daniele
AU - Maggi, Laura
AU - Limatola, Cristina
AU - Di Angelantonio, Silvia
AU - Ragozzino, Davide
ID - 10818
IS - 1
JF - Glia
KW - Cellular and Molecular Neuroscience
KW - Neurology
SN - 0894-1491
TI - Microglia control glutamatergic synapses in the adult mouse hippocampus
VL - 70
ER -
TY - GEN
AB - Complex wiring between neurons underlies the information-processing network enabling all brain functions, including cognition and memory. For understanding how the network is structured, processes information, and changes over time, comprehensive visualization of the architecture of living brain tissue with its cellular and molecular components would open up major opportunities. However, electron microscopy (EM) provides nanometre-scale resolution required for full in-silico reconstruction1–5, yet is limited to fixed specimens and static representations. Light microscopy allows live observation, with super-resolution approaches6–12 facilitating nanoscale visualization, but comprehensive 3D-reconstruction of living brain tissue has been hindered by tissue photo-burden, photobleaching, insufficient 3D-resolution, and inadequate signal-to-noise ratio (SNR). Here we demonstrate saturated reconstruction of living brain tissue. We developed an integrated imaging and analysis technology, adapting stimulated emission depletion (STED) microscopy6,13 in extracellularly labelled tissue14 for high SNR and near-isotropic resolution. Centrally, a two-stage deep-learning approach leveraged previously obtained information on sample structure to drastically reduce photo-burden and enable automated volumetric reconstruction down to single synapse level. Live reconstruction provides unbiased analysis of tissue architecture across time in relation to functional activity and targeted activation, and contextual understanding of molecular labelling. This adoptable technology will facilitate novel insights into the dynamic functional architecture of living brain tissue.
AU - Velicky, Philipp
AU - Miguel Villalba, Eder
AU - Michalska, Julia M
AU - Wei, Donglai
AU - Lin, Zudi
AU - Watson, Jake
AU - Troidl, Jakob
AU - Beyer, Johanna
AU - Ben Simon, Yoav
AU - Sommer, Christoph M
AU - Jahr, Wiebke
AU - Cenameri, Alban
AU - Broichhagen, Johannes
AU - Grant, Seth G. N.
AU - Jonas, Peter M
AU - Novarino, Gaia
AU - Pfister, Hanspeter
AU - Bickel, Bernd
AU - Danzl, Johann G
ID - 11943
T2 - bioRxiv
TI - Saturated reconstruction of living brain tissue
ER -
TY - GEN
AB - Mapping the complex and dense arrangement of cells and their connectivity in brain tissue demands nanoscale spatial resolution imaging. Super-resolution optical microscopy excels at visualizing specific molecules and individual cells but fails to provide tissue context. Here we developed Comprehensive Analysis of Tissues across Scales (CATS), a technology to densely map brain tissue architecture from millimeter regional to nanoscopic synaptic scales in diverse chemically fixed brain preparations, including rodent and human. CATS leverages fixation-compatible extracellular labeling and advanced optical readout, in particular stimulated-emission depletion and expansion microscopy, to comprehensively delineate cellular structures. It enables 3D-reconstructing single synapses and mapping synaptic connectivity by identification and tailored analysis of putative synaptic cleft regions. Applying CATS to the hippocampal mossy fiber circuitry, we demonstrate its power to reveal the system’s molecularly informed ultrastructure across spatial scales and assess local connectivity by reconstructing and quantifying the synaptic input and output structure of identified neurons.
AU - Michalska, Julia M
AU - Lyudchik, Julia
AU - Velicky, Philipp
AU - Korinkova, Hana
AU - Watson, Jake
AU - Cenameri, Alban
AU - Sommer, Christoph M
AU - Venturino, Alessandro
AU - Roessler, Karl
AU - Czech, Thomas
AU - Siegert, Sandra
AU - Novarino, Gaia
AU - Jonas, Peter M
AU - Danzl, Johann G
ID - 11950
T2 - bioRxiv
TI - Uncovering brain tissue architecture across scales with super-resolution light microscopy
ER -
TY - JOUR
AB - Mutations in the chromodomain helicase DNA-binding 8 (CHD8) gene are a frequent cause of autism spectrum disorder (ASD). While its phenotypic spectrum often encompasses macrocephaly, implicating cortical abnormalities, how CHD8 haploinsufficiency affects neurodevelopmental is unclear. Here, employing human cerebral organoids, we find that CHD8 haploinsufficiency disrupted neurodevelopmental trajectories with an accelerated and delayed generation of, respectively, inhibitory and excitatory neurons that yields, at days 60 and 120, symmetrically opposite expansions in their proportions. This imbalance is consistent with an enlargement of cerebral organoids as an in vitro correlate of patients’ macrocephaly. Through an isogenic design of patient-specific mutations and mosaic organoids, we define genotype-phenotype relationships and uncover their cell-autonomous nature. Our results define cell-type-specific CHD8-dependent molecular defects related to an abnormal program of proliferation and alternative splicing. By identifying cell-type-specific effects of CHD8 mutations, our study uncovers reproducible developmental alterations that may be employed for neurodevelopmental disease modeling.
AU - Villa, Carlo Emanuele
AU - Cheroni, Cristina
AU - Dotter, Christoph
AU - López-Tóbon, Alejandro
AU - Oliveira, Bárbara
AU - Sacco, Roberto
AU - Yahya, Aysan Çerağ
AU - Morandell, Jasmin
AU - Gabriele, Michele
AU - Tavakoli, Mojtaba
AU - Lyudchik, Julia
AU - Sommer, Christoph M
AU - Gabitto, Mariano
AU - Danzl, Johann G
AU - Testa, Giuseppe
AU - Novarino, Gaia
ID - 11160
IS - 1
JF - Cell Reports
KW - General Biochemistry
KW - Genetics and Molecular Biology
SN - 2211-1247
TI - CHD8 haploinsufficiency links autism to transient alterations in excitatory and inhibitory trajectories
VL - 39
ER -
TY - THES
AB - Autism spectrum disorders (ASDs) are a group of neurodevelopmental disorders characterized by behavioral symptoms such as problems in social communication and interaction, as
well as repetitive, restricted behaviors and interests. These disorders show a high degree
of heritability and hundreds of risk genes have been identifed using high throughput
sequencing technologies. This genetic heterogeneity has hampered eforts in understanding
the pathogenesis of ASD but at the same time given rise to the concept of convergent
mechanisms. Previous studies have identifed that risk genes for ASD broadly converge
onto specifc functional categories with transcriptional regulation being one of the biggest
groups. In this thesis, I focus on this subgroup of genes and investigate the gene regulatory
consequences of some of them in the context of neurodevelopment.
First, we showed that mutations in the ASD and intellectual disability risk gene Setd5 lead
to perturbations of gene regulatory programs in early cell fate specifcation. In addition,
adult animals display abnormal learning behavior which is mirrored at the transcriptional
level by altered activity dependent regulation of postsynaptic gene expression. Lastly,
we link the regulatory function of Setd5 to its interaction with the Paf1 and the NCoR
complex.
Second, by modeling the heterozygous loss of the top ASD gene CHD8 in human cerebral
organoids we demonstrate profound changes in the developmental trajectories of both
inhibitory and excitatory neurons using single cell RNA-sequencing. While the former
were generated earlier in CHD8+/- organoids, the generation of the latter was shifted to
later times in favor of a prolonged progenitor expansion phase and ultimately increased
organoid size.
Finally, by modeling heterozygous mutations for four ASD associated chromatin modifers,
ASH1L, KDM6B, KMT5B, and SETD5 in human cortical spheroids we show evidence of
regulatory convergence across three of those genes. We observe a shift from dorsal cortical
excitatory neuron fates towards partially ventralized cell types resembling cells from the
lateral ganglionic eminence. As this project is still ongoing at the time of writing, future
experiments will aim at elucidating the regulatory mechanisms underlying this shift with
the aim of linking these three ASD risk genes through biological convergence.
AU - Dotter, Christoph
ID - 12364
SN - 2663-337X
TI - Transcriptional consequences of mutations in genes associated with Autism Spectrum Disorder
ER -
TY - JOUR
AB - Mutations affecting mTOR or RAS signaling underlie defined syndromes (the so-called mTORopathies and RASopathies) with high risk for Autism Spectrum Disorder (ASD). These syndromes show a broad variety of somatic phenotypes including cancers, skin abnormalities, heart disease and facial dysmorphisms. Less well studied are the neuropsychiatric symptoms such as ASD. Here, we assess the relevance of these signalopathies in ASD reviewing genetic, human cell model, rodent studies and clinical trials. We conclude that signalopathies have an increased liability for ASD and that, in particular, ASD individuals with dysmorphic features and intellectual disability (ID) have a higher chance for disruptive mutations in RAS- and mTOR-related genes. Studies on rodent and human cell models confirm aberrant neuronal development as the underlying pathology. Human studies further suggest that multiple hits are necessary to induce the respective phenotypes. Recent clinical trials do only report improvements for comorbid conditions such as epilepsy or cancer but not for behavioral aspects. Animal models show that treatment during early development can rescue behavioral phenotypes. Taken together, we suggest investigating the differential roles of mTOR and RAS signaling in both human and rodent models, and to test drug treatment both during and after neuronal development in the available model systems
AU - Vasic, Verica
AU - Jones, Mattson S.O.
AU - Haslinger, Denise
AU - Knaus, Lisa
AU - Schmeisser, Michael J.
AU - Novarino, Gaia
AU - Chiocchetti, Andreas G.
ID - 10281
IS - 11
JF - Genes
TI - Translating the role of mtor-and ras-associated signalopathies in autism spectrum disorder: Models, mechanisms and treatment
VL - 12
ER -
TY - JOUR
AB - De novo protein synthesis is required for synapse modifications underlying stable memory encoding. Yet neurons are highly compartmentalized cells and how protein synthesis can be regulated at the synapse level is unknown. Here, we characterize neuronal signaling complexes formed by the postsynaptic scaffold GIT1, the mechanistic target of rapamycin (mTOR) kinase, and Raptor that couple synaptic stimuli to mTOR-dependent protein synthesis; and identify NMDA receptors containing GluN3A subunits as key negative regulators of GIT1 binding to mTOR. Disruption of GIT1/mTOR complexes by enhancing GluN3A expression or silencing GIT1 inhibits synaptic mTOR activation and restricts the mTOR-dependent translation of specific activity-regulated mRNAs. Conversely, GluN3A removal enables complex formation, potentiates mTOR-dependent protein synthesis, and facilitates the consolidation of associative and spatial memories in mice. The memory enhancement becomes evident with light or spaced training, can be achieved by selectively deleting GluN3A from excitatory neurons during adulthood, and does not compromise other aspects of cognition such as memory flexibility or extinction. Our findings provide mechanistic insight into synaptic translational control and reveal a potentially selective target for cognitive enhancement.
AU - Conde-Dusman, María J
AU - Dey, Partha N
AU - Elía-Zudaire, Óscar
AU - Garcia Rabaneda, Luis E
AU - García-Lira, Carmen
AU - Grand, Teddy
AU - Briz, Victor
AU - Velasco, Eric R
AU - Andero Galí, Raül
AU - Niñerola, Sergio
AU - Barco, Angel
AU - Paoletti, Pierre
AU - Wesseling, John F
AU - Gardoni, Fabrizio
AU - Tavalin, Steven J
AU - Perez-Otaño, Isabel
ID - 10301
JF - eLife
KW - general immunology and microbiology
KW - general biochemistry
KW - genetics and molecular biology
KW - general medicine
KW - general neuroscience
SN - 2050-084X
TI - Control of protein synthesis and memory by GluN3A-NMDA receptors through inhibition of GIT1/mTORC1 assembly
VL - 10
ER -
TY - JOUR
AB - Chronic psychological stress is one of the most important triggers and environmental risk factors for neuropsychiatric disorders. Chronic stress can influence all organs via the secretion of stress hormones, including glucocorticoids by the adrenal glands, which coordinate the stress response across the body. In the brain, glucocorticoid receptors (GR) are expressed by various cell types including microglia, which are its resident immune cells regulating stress-induced inflammatory processes. To study the roles of microglial GR under normal homeostatic conditions and following chronic stress, we generated a mouse model in which the GR gene is depleted in microglia specifically at adulthood to prevent developmental confounds. We first confirmed that microglia were depleted in GR in our model in males and females among the cingulate cortex and the hippocampus, both stress-sensitive brain regions. Then, cohorts of microglial-GR depleted and wild-type (WT) adult female mice were housed for 3 weeks in a standard or stressful condition, using a chronic unpredictable mild stress (CUMS) paradigm. CUMS induced stress-related behavior in both microglial-GR depleted and WT animals as demonstrated by a decrease of both saccharine preference and progressive ratio breakpoint. Nevertheless, the hippocampal microglial and neural mechanisms underlying the adaptation to stress occurred differently between the two genotypes. Upon CUMS exposure, microglial morphology was altered in the WT controls, without any apparent effect in microglial-GR depleted mice. Furthermore, in the standard environment condition, GR depleted-microglia showed increased expression of pro-inflammatory genes, and genes involved in microglial homeostatic functions (such as Trem2, Cx3cr1 and Mertk). On the contrary, in CUMS condition, GR depleted-microglia showed reduced expression levels of pro-inflammatory genes and increased neuroprotective as well as anti-inflammatory genes compared to WT-microglia. Moreover, in microglial-GR depleted mice, but not in WT mice, CUMS led to a significant reduction of CA1 long-term potentiation and paired-pulse ratio. Lastly, differences in adult hippocampal neurogenesis were observed between the genotypes during normal homeostatic conditions, with microglial-GR deficiency increasing the formation of newborn neurons in the dentate gyrus subgranular zone independently from stress exposure. Together, these findings indicate that, although the deletion of microglial GR did not prevent the animal’s ability to respond to stress, it contributed to modulating hippocampal functions in both standard and stressful conditions, notably by shaping the microglial response to chronic stress.
AU - Picard, Katherine
AU - Bisht, Kanchan
AU - Poggini, Silvia
AU - Garofalo, Stefano
AU - Golia, Maria Teresa
AU - Basilico, Bernadette
AU - Abdallah, Fatima
AU - Ciano Albanese, Naomi
AU - Amrein, Irmgard
AU - Vernoux, Nathalie
AU - Sharma, Kaushik
AU - Hui, Chin Wai
AU - C. Savage, Julie
AU - Limatola, Cristina
AU - Ragozzino, Davide
AU - Maggi, Laura
AU - Branchi, Igor
AU - Tremblay, Marie Ève
ID - 9953
JF - Brain, Behavior, and Immunity
SN - 0889-1591
TI - Microglial-glucocorticoid receptor depletion alters the response of hippocampal microglia and neurons in a chronic unpredictable mild stress paradigm in female mice
VL - 97
ER -
TY - JOUR
AB - P-glycoprotein (ABCB1) and breast cancer resistance protein (ABCG2) restrict at the blood–brain barrier (BBB) the brain distribution of the majority of currently known molecularly targeted anticancer drugs. To improve brain delivery of dual ABCB1/ABCG2 substrates, both ABCB1 and ABCG2 need to be inhibited simultaneously at the BBB. We examined the feasibility of simultaneous ABCB1/ABCG2 inhibition with i.v. co-infusion of erlotinib and tariquidar by studying brain distribution of the model ABCB1/ABCG2 substrate [11C]erlotinib in mice and rhesus macaques with PET. Tolerability of the erlotinib/tariquidar combination was assessed in human embryonic stem cell-derived cerebral organoids. In mice and macaques, baseline brain distribution of [11C]erlotinib was low (brain distribution volume, VT,brain < 0.3 mL/cm3). Co-infusion of erlotinib and tariquidar increased VT,brain in mice by 3.0-fold and in macaques by 3.4- to 5.0-fold, while infusion of erlotinib alone or tariquidar alone led to less pronounced VT,brain increases in both species. Treatment of cerebral organoids with erlotinib/tariquidar led to an induction of Caspase-3-dependent apoptosis. Co-infusion of erlotinib/tariquidar may potentially allow for complete ABCB1/ABCG2 inhibition at the BBB, while simultaneously achieving brain-targeted EGFR inhibition. Our protocol may be applicable to enhance brain delivery of molecularly targeted anticancer drugs for a more effective treatment of brain tumors.
AU - Tournier, N
AU - Goutal, S
AU - Mairinger, S
AU - Lozano, IH
AU - Filip, T
AU - Sauberer, M
AU - Caillé, F
AU - Breuil, L
AU - Stanek, J
AU - Freeman, AF
AU - Novarino, Gaia
AU - Truillet, C
AU - Wanek, T
AU - Langer, O
ID - 8730
IS - 7
JF - Journal of Cerebral Blood Flow and Metabolism
SN - 0271-678x
TI - Complete inhibition of ABCB1 and ABCG2 at the blood-brain barrier by co-infusion of erlotinib and tariquidar to improve brain delivery of the model ABCB1/ABCG2 substrate [11C]erlotinib
VL - 41
ER -
TY - JOUR
AB - De novo loss of function mutations in the ubiquitin ligase-encoding gene Cullin3 lead to autism spectrum disorder (ASD). In mouse, constitutive haploinsufficiency leads to motor coordination deficits as well as ASD-relevant social and cognitive impairments. However, induction of Cul3 haploinsufficiency later in life does not lead to ASD-relevant behaviors, pointing to an important role of Cul3 during a critical developmental window. Here we show that Cul3 is essential to regulate neuronal migration and, therefore, constitutive Cul3 heterozygous mutant mice display cortical lamination abnormalities. At the molecular level, we found that Cul3 controls neuronal migration by tightly regulating the amount of Plastin3 (Pls3), a previously unrecognized player of neural migration. Furthermore, we found that Pls3 cell-autonomously regulates cell migration by regulating actin cytoskeleton organization, and its levels are inversely proportional to neural migration speed. Finally, we provide evidence that cellular phenotypes associated with autism-linked gene haploinsufficiency can be rescued by transcriptional activation of the intact allele in vitro, offering a proof of concept for a potential therapeutic approach for ASDs.
AU - Morandell, Jasmin
AU - Schwarz, Lena A
AU - Basilico, Bernadette
AU - Tasciyan, Saren
AU - Dimchev, Georgi A
AU - Nicolas, Armel
AU - Sommer, Christoph M
AU - Kreuzinger, Caroline
AU - Dotter, Christoph
AU - Knaus, Lisa
AU - Dobler, Zoe
AU - Cacci, Emanuele
AU - Schur, Florian KM
AU - Danzl, Johann G
AU - Novarino, Gaia
ID - 9429
IS - 1
JF - Nature Communications
KW - General Biochemistry
KW - Genetics and Molecular Biology
TI - Cul3 regulates cytoskeleton protein homeostasis and cell migration during a critical window of brain development
VL - 12
ER -
TY - JOUR
AB - In recent years, many genes have been associated with chromatinopathies classified as “Cornelia de Lange Syndrome‐like.” It is known that the phenotype of these patients becomes less recognizable, overlapping to features characteristic of other syndromes caused by genetic variants affecting different regulators of chromatin structure and function. Therefore, Cornelia de Lange syndrome diagnosis might be arduous due to the seldom discordance between unexpected molecular diagnosis and clinical evaluation. Here, we review the molecular features of Cornelia de Lange syndrome, supporting the hypothesis that “CdLS‐like syndromes” are part of a larger “rare disease family” sharing multiple clinical features and common disrupted molecular pathways.
AU - Avagliano, Laura
AU - Parenti, Ilaria
AU - Grazioli, Paolo
AU - Di Fede, Elisabetta
AU - Parodi, Chiara
AU - Mariani, Milena
AU - Kaiser, Frank J.
AU - Selicorni, Angelo
AU - Gervasini, Cristina
AU - Massa, Valentina
ID - 7149
IS - 1
JF - Clinical Genetics
SN - 0009-9163
TI - Chromatinopathies: A focus on Cornelia de Lange syndrome
VL - 97
ER -
TY - JOUR
AB - Characteristic or classic phenotype of Cornelia de Lange syndrome (CdLS) is associated with a recognisable facial pattern. However, the heterogeneity in causal genes and the presence of overlapping syndromes have made it increasingly difficult to diagnose only by clinical features. DeepGestalt technology, and its app Face2Gene, is having a growing impact on the diagnosis and management of genetic diseases by analysing the features of affected individuals. Here, we performed a phenotypic study on a cohort of 49 individuals harbouring causative variants in known CdLS genes in order to evaluate Face2Gene utility and sensitivity in the clinical diagnosis of CdLS. Based on the profile images of patients, a diagnosis of CdLS was within the top five predicted syndromes for 97.9% of our cases and even listed as first prediction for 83.7%. The age of patients did not seem to affect the prediction accuracy, whereas our results indicate a correlation between the clinical score and affected genes. Furthermore, each gene presents a different pattern recognition that may be used to develop new neural networks with the goal of separating different genetic subtypes in CdLS. Overall, we conclude that computer-assisted image analysis based on deep learning could support the clinical diagnosis of CdLS.
AU - Latorre-Pellicer, Ana
AU - Ascaso, Ángela
AU - Trujillano, Laura
AU - Gil-Salvador, Marta
AU - Arnedo, Maria
AU - Lucia-Campos, Cristina
AU - Antoñanzas-Pérez, Rebeca
AU - Marcos-Alcalde, Iñigo
AU - Parenti, Ilaria
AU - Bueno-Lozano, Gloria
AU - Musio, Antonio
AU - Puisac, Beatriz
AU - Kaiser, Frank J.
AU - Ramos, Feliciano J.
AU - Gómez-Puertas, Paulino
AU - Pié, Juan
ID - 7488
IS - 3
JF - International Journal of Molecular Sciences
SN - 16616596
TI - Evaluating Face2Gene as a tool to identify Cornelia de Lange syndrome by facial phenotypes
VL - 21
ER -
TY - JOUR
AB - CLC chloride/proton exchangers may support acidification of endolysosomes and raise their luminal Cl− concentration. Disruption of endosomal ClC‐3 causes severe neurodegeneration. To assess the importance of ClC‐3 Cl−/H+ exchange, we now generate Clcn3unc/unc mice in which ClC‐3 is converted into a Cl− channel. Unlike Clcn3−/− mice, Clcn3unc/unc mice appear normal owing to compensation by ClC‐4 with which ClC‐3 forms heteromers. ClC‐4 protein levels are strongly reduced in Clcn3−/−, but not in Clcn3unc/unc mice because ClC‐3unc binds and stabilizes ClC‐4 like wild‐type ClC‐3. Although mice lacking ClC‐4 appear healthy, its absence in Clcn3unc/unc/Clcn4−/− mice entails even stronger neurodegeneration than observed in Clcn3−/− mice. A fraction of ClC‐3 is found on synaptic vesicles, but miniature postsynaptic currents and synaptic vesicle acidification are not affected in Clcn3unc/unc or Clcn3−/− mice before neurodegeneration sets in. Both, Cl−/H+‐exchange activity and the stabilizing effect on ClC‐4, are central to the biological function of ClC‐3.
AU - Weinert, Stefanie
AU - Gimber, Niclas
AU - Deuschel, Dorothea
AU - Stuhlmann, Till
AU - Puchkov, Dmytro
AU - Farsi, Zohreh
AU - Ludwig, Carmen F.
AU - Novarino, Gaia
AU - López-Cayuqueo, Karen I.
AU - Planells-Cases, Rosa
AU - Jentsch, Thomas J.
ID - 7586
JF - EMBO Journal
SN - 02614189
TI - Uncoupling endosomal CLC chloride/proton exchange causes severe neurodegeneration
VL - 39
ER -
TY - JOUR
AB - The NIPBL/MAU2 heterodimer loads cohesin onto chromatin. Mutations inNIPBLaccount for most cases ofthe rare developmental disorder Cornelia de Lange syndrome (CdLS). Here we report aMAU2 variant causing CdLS, a deletion of seven amino acids that impairs the interaction between MAU2 and the NIPBL N terminus.Investigating this interaction, we discovered that MAU2 and the NIPBL N terminus are largely dispensable fornormal cohesin and NIPBL function in cells with a NIPBL early truncating mutation. Despite a predicted fataloutcome of an out-of-frame single nucleotide duplication inNIPBL, engineered in two different cell lines,alternative translation initiation yields a form of NIPBL missing N-terminal residues. This form cannot interactwith MAU2, but binds DNA and mediates cohesin loading. Altogether, our work reveals that cohesin loading can occur independently of functional NIPBL/MAU2 complexes and highlights a novel mechanism protectiveagainst out-of-frame mutations that is potentially relevant for other genetic conditions.
AU - Parenti, Ilaria
AU - Diab, Farah
AU - Gil, Sara Ruiz
AU - Mulugeta, Eskeatnaf
AU - Casa, Valentina
AU - Berutti, Riccardo
AU - Brouwer, Rutger W.W.
AU - Dupé, Valerie
AU - Eckhold, Juliane
AU - Graf, Elisabeth
AU - Puisac, Beatriz
AU - Ramos, Feliciano
AU - Schwarzmayr, Thomas
AU - Gines, Macarena Moronta
AU - Van Staveren, Thomas
AU - Van Ijcken, Wilfred F.J.
AU - Strom, Tim M.
AU - Pié, Juan
AU - Watrin, Erwan
AU - Kaiser, Frank J.
AU - Wendt, Kerstin S.
ID - 7877
IS - 7
JF - Cell Reports
TI - MAU2 and NIPBL variants impair the heterodimerization of the cohesin loader subunits and cause Cornelia de Lange syndrome
VL - 31
ER -
TY - JOUR
AB - Neurodevelopmental disorders (NDDs) are a class of disorders affecting brain development and function and are characterized by wide genetic and clinical variability. In this review, we discuss the multiple factors that influence the clinical presentation of NDDs, with particular attention to gene vulnerability, mutational load, and the two-hit model. Despite the complex architecture of
mutational events associated with NDDs, the various proteins involved appear to converge on common pathways, such as synaptic plasticity/function, chromatin remodelers and the mammalian target of rapamycin (mTOR) pathway. A thorough understanding of the mechanisms behind these pathways will hopefully lead to the identification of candidates that could be targeted for treatment approaches.
AU - Parenti, Ilaria
AU - Garcia Rabaneda, Luis E
AU - Schön, Hanna
AU - Novarino, Gaia
ID - 7957
IS - 8
JF - Trends in Neurosciences
SN - 01662236
TI - Neurodevelopmental disorders: From genetics to functional pathways
VL - 43
ER -
TY - THES
AB - The development of the human brain occurs through a tightly regulated series of dynamic and adaptive processes during prenatal and postnatal life. A disruption of this strictly orchestrated series of events can lead to a number of neurodevelopmental conditions, including Autism Spectrum Disorders (ASDs). ASDs are a very common, etiologically and phenotypically heterogeneous group of disorders sharing the core symptoms of social interaction and communication deficits and restrictive and repetitive interests and behaviors. They are estimated to affect one in 59 individuals in the U.S. and, over the last three decades, mutations in more than a hundred genetic loci have been convincingly linked to ASD pathogenesis. Yet, for the vast majority of these ASD-risk genes their role during brain development and precise molecular function still remain elusive.
De novo loss of function mutations in the ubiquitin ligase-encoding gene Cullin 3 (CUL3) lead to ASD. In the study described here, we used Cul3 mouse models to evaluate the consequences of Cul3 mutations in vivo. Our results show that Cul3 heterozygous knockout mice exhibit deficits in motor coordination as well as ASD-relevant social and cognitive impairments. Cul3+/-, Cul3+/fl Emx1-Cre and Cul3fl/fl Emx1-Cre mutant brains display cortical lamination abnormalities due to defective migration of post-mitotic excitatory neurons, as well as reduced numbers of excitatory and inhibitory neurons. In line with the observed abnormal cortical organization, Cul3 heterozygous deletion is associated with decreased spontaneous excitatory and inhibitory activity in the cortex. At the molecular level we show that Cul3 regulates cytoskeletal and adhesion protein abundance in the mouse embryonic cortex. Abnormal regulation of cytoskeletal proteins in Cul3 mutant neural cells results in atypical organization of the actin mesh at the cell leading edge. Of note, heterozygous deletion of Cul3 in adult mice does not induce the majority of the behavioral defects observed in constitutive Cul3 haploinsufficient animals, pointing to a critical time-window for Cul3 deficiency.
In conclusion, our data indicate that Cul3 plays a critical role in the regulation of cytoskeletal proteins and neuronal migration. ASD-associated defects and behavioral abnormalities are primarily due to dosage sensitive Cul3 functions at early brain developmental stages.
AU - Morandell, Jasmin
ID - 8620
SN - 2663-337X
TI - Illuminating the role of Cul3 in autism spectrum disorder pathogenesis
ER -
TY - GEN
AB - De novo loss of function mutations in the ubiquitin ligase-encoding gene Cullin3 (CUL3) lead to autism spectrum disorder (ASD). Here, we used Cul3 mouse models to evaluate the consequences of Cul3 mutations in vivo. Our results show that Cul3 haploinsufficient mice exhibit deficits in motor coordination as well as ASD-relevant social and cognitive impairments. Cul3 mutant brain displays cortical lamination abnormalities due to defective neuronal migration and reduced numbers of excitatory and inhibitory neurons. In line with the observed abnormal columnar organization, Cul3 haploinsufficiency is associated with decreased spontaneous excitatory and inhibitory activity in the cortex. At the molecular level, employing a quantitative proteomic approach, we show that Cul3 regulates cytoskeletal and adhesion protein abundance in mouse embryos. Abnormal regulation of cytoskeletal proteins in Cul3 mutant neuronal cells results in atypical organization of the actin mesh at the cell leading edge, likely causing the observed migration deficits. In contrast to these important functions early in development, Cul3 deficiency appears less relevant at adult stages. In fact, induction of Cul3 haploinsufficiency in adult mice does not result in the behavioral defects observed in constitutive Cul3 haploinsufficient animals. Taken together, our data indicate that Cul3 has a critical role in the regulation of cytoskeletal proteins and neuronal migration and that ASD-associated defects and behavioral abnormalities are primarily due to Cul3 functions at early developmental stages.
AU - Morandell, Jasmin
AU - Schwarz, Lena A
AU - Basilico, Bernadette
AU - Tasciyan, Saren
AU - Nicolas, Armel
AU - Sommer, Christoph M
AU - Kreuzinger, Caroline
AU - Knaus, Lisa
AU - Dobler, Zoe
AU - Cacci, Emanuele
AU - Danzl, Johann G
AU - Novarino, Gaia
ID - 7800
T2 - bioRxiv
TI - Cul3 regulates cytoskeleton protein homeostasis and cell migration during a critical window of brain development
ER -
TY - JOUR
AB - The possibility to generate construct valid animal models enabled the development and testing of therapeutic strategies targeting the core features of autism spectrum disorders (ASDs). At the same time, these studies highlighted the necessity of identifying sensitive developmental time windows for successful therapeutic interventions. Animal and human studies also uncovered the possibility to stratify the variety of ASDs in molecularly distinct subgroups, potentially facilitating effective treatment design. Here, we focus on the molecular pathways emerging as commonly affected by mutations in diverse ASD-risk genes, on their role during critical windows of brain development and the potential treatments targeting these biological processes.
AU - Basilico, Bernadette
AU - Morandell, Jasmin
AU - Novarino, Gaia
ID - 8131
IS - 12
JF - Current Opinion in Genetics and Development
SN - 0959437X
TI - Molecular mechanisms for targeted ASD treatments
VL - 65
ER -
TY - JOUR
AB - Clinical Utility Gene Card. 1. Name of Disease (Synonyms): Pontocerebellar hypoplasia type 9 (PCH9) and spastic paraplegia-63 (SPG63). 2. OMIM# of the Disease: 615809 and 615686. 3. Name of the Analysed Genes or DNA/Chromosome Segments: AMPD2 at 1p13.3. 4. OMIM# of the Gene(s): 102771.
AU - Marsh, Ashley
AU - Novarino, Gaia
AU - Lockhart, Paul
AU - Leventer, Richard
ID - 105
JF - European Journal of Human Genetics
TI - CUGC for pontocerebellar hypoplasia type 9 and spastic paraplegia-63
VL - 27
ER -
TY - JOUR
AB - P-Glycoprotein (ABCB1) and breast cancer resistance protein (ABCG2) are two efflux transporters at the blood–brain barrier (BBB), which effectively restrict brain distribution of diverse drugs, such as tyrosine kinase inhibitors. There is a crucial need for pharmacological ABCB1 and ABCG2 inhibition protocols for a more effective treatment of brain diseases. In the present study, seven marketed drugs (osimertinib, erlotinib, nilotinib, imatinib, lapatinib, pazopanib, and cyclosporine A) and one nonmarketed drug (tariquidar), with known in vitro ABCB1/ABCG2 inhibitory properties, were screened for their inhibitory potency at the BBB in vivo. Positron emission tomography (PET) using the model ABCB1/ABCG2 substrate [11C]erlotinib was performed in mice. Tested inhibitors were administered as i.v. bolus injections at 30 min before the start of the PET scan, followed by a continuous i.v. infusion for the duration of the PET scan. Five of the tested drugs increased total distribution volume of [11C]erlotinib in the brain (VT,brain) compared to vehicle-treated animals (tariquidar, + 69%; erlotinib, + 19% and +23% for the 21.5 mg/kg and the 43 mg/kg dose, respectively; imatinib, + 22%; lapatinib, + 25%; and cyclosporine A, + 49%). For all drugs, increases in [11C]erlotinib brain distribution were lower than in Abcb1a/b(−/−)Abcg2(−/−) mice (+149%), which suggested that only partial ABCB1/ABCG2 inhibition was reached at the mouse BBB. The plasma concentrations of the tested drugs at the time of the PET scan were higher than clinically achievable plasma concentrations. Some of the tested drugs led to significant increases in blood radioactivity concentrations measured at the end of the PET scan (erlotinib, + 103% and +113% for the 21.5 mg/kg and the 43 mg/kg dose, respectively; imatinib, + 125%; and cyclosporine A, + 101%), which was most likely caused by decreased hepatobiliary excretion of radioactivity. Taken together, our data suggest that some marketed tyrosine kinase inhibitors may be repurposed to inhibit ABCB1 and ABCG2 at the BBB. From a clinical perspective, moderate increases in brain delivery despite the administration of high i.v. doses as well as peripheral drug–drug interactions due to transporter inhibition in clearance organs question the translatability of this concept.
AU - Traxl, Alexander
AU - Mairinger, Severin
AU - Filip, Thomas
AU - Sauberer, Michael
AU - Stanek, Johann
AU - Poschner, Stefan
AU - Jäger, Walter
AU - Zoufal, Viktoria
AU - Novarino, Gaia
AU - Tournier, Nicolas
AU - Bauer, Martin
AU - Wanek, Thomas
AU - Langer, Oliver
ID - 6088
IS - 3
JF - Molecular Pharmaceutics
TI - Inhibition of ABCB1 and ABCG2 at the mouse blood-brain barrier with marketed drugs to improve brain delivery of the model ABCB1/ABCG2 substrate [11C]erlotinib
VL - 16
ER -
TY - JOUR
AB - Investigating neuronal activity using genetically encoded Ca2+ indicators in behaving animals is hampered by inaccuracies in spike inference from fluorescent tracers. Here we combine two‐photon [Ca2+] imaging with cell‐attached recordings, followed by post hoc determination of the expression level of GCaMP6f, to explore how it affects the amplitude, kinetics and temporal summation of somatic [Ca2+] transients in mouse hippocampal pyramidal cells (PCs). The amplitude of unitary [Ca2+] transients (evoked by a single action potential) negatively correlates with GCaMP6f expression, but displays large variability even among PCs with similarly low expression levels. The summation of fluorescence signals is frequency‐dependent, supralinear and also shows remarkable cell‐to‐cell variability. We performed experimental data‐based simulations and found that spike inference error rates using MLspike depend strongly on unitary peak amplitudes and GCaMP6f expression levels. We provide simple methods for estimating the unitary [Ca2+] transients in individual weakly GCaMP6f‐expressing PCs, with which we achieve spike inference error rates of ∼5%.
AU - Éltes, Tímea
AU - Szoboszlay, Miklos
AU - Szigeti, Margit Katalin
AU - Nusser, Zoltan
ID - 6470
IS - 11
JF - Journal of Physiology
SN - 00223751
TI - Improved spike inference accuracy by estimating the peak amplitude of unitary [Ca2+] transients in weakly GCaMP6f-expressing hippocampal pyramidal cells
VL - 597
ER -
TY - JOUR
AB - Until recently, a great amount of brain studies have been conducted in human post mortem tissues, cell lines and model organisms. These researches provided useful insights regarding cell-cell interactions occurring in the brain. However, such approaches suffer from technical limitations and inaccurate modeling of the tissue 3D cytoarchitecture. Importantly, they might lack a human genetic background essential for disease modeling. With the development of protocols to generate human cerebral organoids, we are now closer to reproducing the early stages of human brain development in vitro. As a result, more relevant cell-cell interaction studies can be conducted.
In this review, we discuss the advantages of 3D cultures over 2D in modulating brain cell-cell interactions during physiological and pathological development, as well as the progress made in developing organoids in which neurons, macroglia, microglia and vascularization are present. Finally, we debate the limitations of those models and possible future directions.
AU - Oliveira, Bárbara
AU - Yahya, Aysan Çerağ
AU - Novarino, Gaia
ID - 6896
JF - Brain Research
SN - 00068993
TI - Modeling cell-cell interactions in the brain using cerebral organoids
VL - 1724
ER -
TY - JOUR
AU - Morandell, Jasmin
AU - Nicolas, Armel
AU - Schwarz, Lena A
AU - Novarino, Gaia
ID - 7415
IS - Supplement 6
JF - European Neuropsychopharmacology
SN - 0924-977X
TI - S.16.05 Illuminating the role of the e3 ubiquitin ligase cullin3 in brain development and autism
VL - 29
ER -
TY - JOUR
AU - Knaus, Lisa
AU - Tarlungeanu, Dora-Clara
AU - Novarino, Gaia
ID - 7414
IS - Supplement 6
JF - European Neuropsychopharmacology
SN - 0924-977X
TI - S.16.03 A homozygous missense mutation in SLC7A5 leads to autism spectrum disorder and microcephaly
VL - 29
ER -
TY - JOUR
AB - Inhibition of the endoplasmic reticulum stress pathway may hold the key to Zika virus-associated microcephaly treatment.
AU - Novarino, Gaia
ID - 456
IS - 423
JF - Science Translational Medicine
TI - Zika-associated microcephaly: Reduce the stress and race for the treatment
VL - 10
ER -
TY - JOUR
AB - Despite the remarkable number of scientific breakthroughs of the last 100 years, the treatment of neurodevelopmental
disorders (e.g., autism spectrum disorder, intellectual disability) remains a great challenge. Recent advancements in
genomics, such as whole-exome or whole-genome sequencing, have enabled scientists to identify numerous
mutations underlying neurodevelopmental disorders. Given the few hundred risk genes that have been discovered,
the etiological variability and the heterogeneous clinical presentation, the need for genotype — along with phenotype-
based diagnosis of individual patients has become a requisite. In this review we look at recent advancements in
genomic analysis and their translation into clinical practice.
AU - Tarlungeanu, Dora-Clara
AU - Novarino, Gaia
ID - 5888
IS - 8
JF - Experimental & Molecular Medicine
SN - 2092-6413
TI - Genomics in neurodevelopmental disorders: an avenue to personalized medicine
VL - 50
ER -
TY - JOUR
AB - The precise control of neural stem cell (NSC) proliferation and differentiation is crucial for the development and function of the human brain. Here, we review the emerging links between the alteration of embryonic and adult neurogenesis and the etiology of neuropsychiatric disorders (NPDs) such as autism spectrum disorders (ASDs) and schizophrenia (SCZ), as well as the advances in stem cell-based modeling and the novel therapeutic targets derived from these studies.
AU - Sacco, Roberto
AU - Cacci, Emanuele
AU - Novarino, Gaia
ID - 546
IS - 2
JF - Current Opinion in Neurobiology
TI - Neural stem cells in neuropsychiatric disorders
VL - 48
ER -
TY - JOUR
AB - Background: Transport protein particle (TRAPP) is a multisubunit complex that regulates membrane trafficking through the Golgi apparatus. The clinical phenotype associated with mutations in various TRAPP subunits has allowed elucidation of their functions in specific tissues. The role of some subunits in human disease, however, has not been fully established, and their functions remain uncertain.
Objective: We aimed to expand the range of neurodevelopmental disorders associated with mutations in TRAPP subunits by exome sequencing of consanguineous families.
Methods: Linkage and homozygosity mapping and candidate gene analysis were used to identify homozygous mutations in families. Patient fibroblasts were used to study splicing defect and zebrafish to model the disease.
Results: We identified six individuals from three unrelated families with a founder homozygous splice mutation in TRAPPC6B, encoding a core subunit of the complex TRAPP I. Patients manifested a neurodevelopmental disorder characterised by microcephaly, epilepsy and autistic features, and showed splicing defect. Zebrafish trappc6b morphants replicated the human phenotype, displaying decreased head size and neuronal hyperexcitability, leading to a lower seizure threshold.
Conclusion: This study provides clinical and functional evidence of the role of TRAPPC6B in brain development and function.
AU - Marin Valencia, Isaac
AU - Novarino, Gaia
AU - Johansen, Anide
AU - Rosti, Başak
AU - Issa, Mahmoud
AU - Musaev, Damir
AU - Bhat, Gifty
AU - Scott, Eric
AU - Silhavy, Jennifer
AU - Stanley, Valentina
AU - Rosti, Rasim
AU - Gleeson, Jeremy
AU - Imam, Farhad
AU - Zaki, Maha
AU - Gleeson, Joseph
ID - 691
IS - 1
JF - Journal of Medical Genetics
SN - 0022-2593
TI - A homozygous founder mutation in TRAPPC6B associates with a neurodevelopmental disorder characterised by microcephaly epilepsy and autistic features
VL - 55
ER -
TY - THES
AB - Autism spectrum disorders (ASD) are a group of genetic disorders often overlapping with other neurological conditions. Despite the remarkable number of scientific breakthroughs of the last 100 years, the treatment of neurodevelopmental disorders (e.g. autism spectrum disorder, intellectual disability, epilepsy) remains a great challenge. Recent advancements in geno mics, like whole-exome or whole-genome sequencing, have enabled scientists to identify numerous mutations underlying neurodevelopmental disorders. Given the few hundred risk genes that were discovered, the etiological variability and the heterogeneous phenotypic outcomes, the need for genotype -along with phenotype- based diagnosis of individual patients becomes a requisite. Driven by this rationale, in a previous study our group described mutations, identified via whole - exome sequencing, in the gene BCKDK – encoding for a key regulator of branched chain amin o acid (BCAA) catabolism - as a cause of ASD. Following up on the role of BCAAs, in the study described here we show that the solute carrier transporter 7a5 (SLC7A5), a large neutral amino acid transporter localized mainly at the blood brain barrier (BBB), has an essential role in maintaining normal levels of brain BCAAs. In mice, deletion of Slc7a5 from the endothelial cells of the BBB leads to atypical brain amino acid profile, abnormal mRNA translation and severe neurolo gical abnormalities. Additionally, deletion of Slc7a5 from the neural progenitor cell population leads to microcephaly. Interestingly, we demonstrate that BCAA intracerebroventricular administration ameliorates abnormal behaviors in adult mutant mice. Furthermore, whole - exome sequencing of patients diagnosed with neurological dis o r ders helped us identify several patients with autistic traits, microcephaly and motor delay carrying deleterious homozygous mutations in the SLC7A5 gene. In conclusion, our data elucidate a neurological syndrome defined by SLC7A5 mutations and support an essential role for t he BCAA s in human bra in function. Together with r ecent studies (described in chapter two) that have successfully made the transition into clinical practice, our findings on the role of B CAAs might have a crucial impact on the development of novel individualized therapeutic strategies for ASD.
AU - Tarlungeanu, Dora-Clara
ID - 395
SN - 2663-337X
TI - The branched chain amino acids in autism spectrum disorders
ER -
TY - JOUR
AB - SETD5 gene mutations have been identified as a frequent cause of idiopathic intellectual disability. Here we show that Setd5-haploinsufficient mice present developmental defects such as abnormal brain-to-body weight ratios and neural crest defect-associated phenotypes. Furthermore, Setd5-mutant mice show impairments in cognitive tasks, enhanced long-term potentiation, delayed ontogenetic profile of ultrasonic vocalization, and behavioral inflexibility. Behavioral issues are accompanied by abnormal expression of postsynaptic density proteins previously associated with cognition. Our data additionally indicate that Setd5 regulates RNA polymerase II dynamics and gene transcription via its interaction with the Hdac3 and Paf1 complexes, findings potentially explaining the gene expression defects observed in Setd5-haploinsufficient mice. Our results emphasize the decisive role of Setd5 in a biological pathway found to be disrupted in humans with intellectual disability and autism spectrum disorder.
AU - Deliu, Elena
AU - Arecco, Niccoló
AU - Morandell, Jasmin
AU - Dotter, Christoph
AU - Contreras, Ximena
AU - Girardot, Charles
AU - Käsper, Eva
AU - Kozlova, Alena
AU - Kishi, Kasumi
AU - Chiaradia, Ilaria
AU - Noh, Kyung
AU - Novarino, Gaia
ID - 3
IS - 12
JF - Nature Neuroscience
TI - Haploinsufficiency of the intellectual disability gene SETD5 disturbs developmental gene expression and cognition
VL - 21
ER -
TY - JOUR
AB - RNA-dependent RNA polymerases (RdRps) play a key role in the life cycle of RNA viruses and impact their immunobiology. The arenavirus lymphocytic choriomeningitis virus (LCMV) strain Clone 13 provides a benchmark model for studying chronic infection. A major genetic determinant for its ability to persist maps to a single amino acid exchange in the viral L protein, which exhibits RdRp activity, yet its functional consequences remain elusive. To unravel the L protein interactions with the host proteome, we engineered infectious L protein-tagged LCMV virions by reverse genetics. A subsequent mass-spectrometric analysis of L protein pulldowns from infected human cells revealed a comprehensive network of interacting host proteins. The obtained LCMV L protein interactome was bioinformatically integrated with known host protein interactors of RdRps from other RNA viruses, emphasizing interconnected modules of human proteins. Functional characterization of selected interactors highlighted proviral (DDX3X) as well as antiviral (NKRF, TRIM21) host factors. To corroborate these findings, we infected Trim21-/-mice with LCMV and found impaired virus control in chronic infection. These results provide insights into the complex interactions of the arenavirus LCMV and other viral RdRps with the host proteome and contribute to a better molecular understanding of how chronic viruses interact with their host.
AU - Khamina, Kseniya
AU - Lercher, Alexander
AU - Caldera, Michael
AU - Schliehe, Christopher
AU - Vilagos, Bojan
AU - Sahin, Mehmet
AU - Kosack, Lindsay
AU - Bhattacharya, Anannya
AU - Májek, Peter
AU - Stukalov, Alexey
AU - Sacco, Roberto
AU - James, Leo
AU - Pinschewer, Daniel
AU - Bennett, Keiryn
AU - Menche, Jörg
AU - Bergthaler, Andreas
ID - 540
IS - 12
JF - PLoS Pathogens
SN - 15537366
TI - Characterization of host proteins interacting with the lymphocytic choriomeningitis virus L protein
VL - 13
ER -
TY - CHAP
AB - Genetic factors might be largely responsible for the development of autism spectrum disorder (ASD) that alone or in combination with specific environmental risk factors trigger the pathology. Multiple mutations identified in ASD patients that impair synaptic function in the central nervous system are well studied in animal models. How these mutations might interact with other risk factors is not fully understood though. Additionally, how systems outside of the brain are altered in the context of ASD is an emerging area of research. Extracerebral influences on the physiology could begin in utero and contribute to changes in the brain and in the development of other body systems and further lead to epigenetic changes. Therefore, multiple recent studies have aimed at elucidating the role of gene-environment interactions in ASD. Here we provide an overview on the extracerebral systems that might play an important associative role in ASD and review evidence regarding the potential roles of inflammation, trace metals, metabolism, genetic susceptibility, enteric nervous system function and the microbiota of the gastrointestinal (GI) tract on the development of endophenotypes in animal models of ASD. By influencing environmental conditions, it might be possible to reduce or limit the severity of ASD pathology.
AU - Hill Yardin, Elisa
AU - Mckeown, Sonja
AU - Novarino, Gaia
AU - Grabrucker, Andreas
ED - Schmeisser, Michael
ED - Boekers, Tobias
ID - 623
SN - 03015556
T2 - Translational Anatomy and Cell Biology of Autism Spectrum Disorder
TI - Extracerebral dysfunction in animal models of autism spectrum disorder
VL - 224
ER -
TY - CHAP
AB - As autism spectrum disorder (ASD) is largely regarded as a neurodevelopmental condition, long-time consensus was that its hallmark features are irreversible. However, several studies from recent years using defined mouse models of ASD have provided clear evidence that in mice neurobiological and behavioural alterations can be ameliorated or even reversed by genetic restoration or pharmacological treatment either before or after symptom onset. Here, we review findings on genetic and pharmacological reversibility of phenotypes in mouse models of ASD. Our review should give a comprehensive overview on both aspects and encourage future studies to better understand the underlying molecular mechanisms that might be translatable from animals to humans.
AU - Schroeder, Jan
AU - Deliu, Elena
AU - Novarino, Gaia
AU - Schmeisser, Michael
ED - Schmeisser, Michael
ED - Boekers, Tobias
ID - 634
T2 - Translational Anatomy and Cell Biology of Autism Spectrum Disorder
TI - Genetic and pharmacological reversibility of phenotypes in mouse models of autism spectrum disorder
VL - 224
ER -
TY - JOUR
AB - Human neurons transplanted into a mouse model for Alzheimer’s disease show human-specific vulnerability to β-amyloid plaques and may help to identify new therapeutic targets.
AU - Novarino, Gaia
ID - 656
IS - 381
JF - Science Translational Medicine
SN - 19466234
TI - Modeling Alzheimer's disease in mice with human neurons
VL - 9
ER -
TY - JOUR
AB - Perinatal exposure to penicillin may result in longlasting gut and behavioral changes.
AU - Novarino, Gaia
ID - 667
IS - 387
JF - Science Translational Medicine
SN - 19466234
TI - The antisocial side of antibiotics
VL - 9
ER -
TY - JOUR
AB - Rett syndrome modeling in monkey mirrors the human disorder.
AU - Novarino, Gaia
ID - 689
IS - 393
JF - Science Translational Medicine
SN - 19466234
TI - Rett syndrome modeling goes simian
VL - 9
ER -
TY - JOUR
AB - Leading autism-associated mutation in mouse partially mimics human disorder.
AU - Novarino, Gaia
ID - 702
IS - 399
JF - Science Translational Medicine
SN - 19466234
TI - The riddle of CHD8 haploinsufficiency in autism spectrum disorder
VL - 9
ER -
TY - JOUR
AB - To determine the dynamics of allelic-specific expression during mouse development, we analyzed RNA-seq data from 23 F1 tissues from different developmental stages, including 19 female tissues allowing X chromosome inactivation (XCI) escapers to also be detected. We demonstrate that allelic expression arising from genetic or epigenetic differences is highly tissue-specific. We find that tissue-specific strain-biased gene expression may be regulated by tissue-specific enhancers or by post-transcriptional differences in stability between the alleles. We also find that escape from X-inactivation is tissue-specific, with leg muscle showing an unexpectedly high rate of XCI escapers. By surveying a range of tissues during development, and performing extensive validation, we are able to provide a high confidence list of mouse imprinted genes including 18 novel genes. This shows that cluster size varies dynamically during development and can be substantially larger than previously thought, with the Igf2r cluster extending over 10 Mb in placenta.
AU - Andergassen, Daniel
AU - Dotter, Christoph
AU - Wenzel, Dyniel
AU - Sigl, Verena
AU - Bammer, Philipp
AU - Muckenhuber, Markus
AU - Mayer, Daniela
AU - Kulinski, Tomasz
AU - Theussl, Hans
AU - Penninger, Josef
AU - Bock, Christoph
AU - Barlow, Denise
AU - Pauler, Florian
AU - Hudson, Quanah
ID - 713
JF - eLife
SN - 2050084X
TI - Mapping the mouse Allelome reveals tissue specific regulation of allelic expression
VL - 6
ER -
TY - JOUR
AB - Background HIV-1 infection and drug abuse are frequently co-morbid and their association greatly increases the severity of HIV-1-induced neuropathology. While nucleus accumbens (NAcc) function is severely perturbed by drugs of abuse, little is known about how HIV-1 infection affects NAcc. Methods We used calcium and voltage imaging to investigate the effect of HIV-1 trans-activator of transcription (Tat) on rat NAcc. Based on previous neuronal studies, we hypothesized that Tat modulates intracellular Ca2+ homeostasis of NAcc neurons. Results We provide evidence that Tat triggers a Ca2+ signaling cascade in NAcc medium spiny neurons (MSN) expressing D1-like dopamine receptors leading to neuronal depolarization. Firstly, Tat induced inositol 1,4,5-trisphsophate (IP3) receptor-mediated Ca2+ release from endoplasmic reticulum, followed by Ca2+ and Na+ influx via transient receptor potential canonical channels. The influx of cations depolarizes the membrane promoting additional Ca2+ entry through voltage-gated P/Q-type Ca2+ channels and opening of tetrodotoxin-sensitive Na+ channels. By activating this mechanism, Tat elicits a feed-forward depolarization increasing the excitability of D1-phosphatidylinositol-linked NAcc MSN. We previously found that cocaine targets NAcc neurons directly (independent of the inhibition of dopamine transporter) only when IP3-generating mechanisms are concomitantly initiated. When tested here, cocaine produced a dose-dependent potentiation of the effect of Tat on cytosolic Ca2+. Conclusion We describe for the first time a HIV-1 Tat-triggered Ca2+ signaling in MSN of NAcc involving TRPC and depolarization and a potentiation of the effect of Tat by cocaine, which may be relevant for the reward axis in cocaine-abusing HIV-1-positive patients.
AU - Brailoiu, Gabriela
AU - Deliu, Elena
AU - Barr, Jeffrey
AU - Console Bram, Linda
AU - Ciuciu, Alexandra
AU - Abood, Mary
AU - Unterwald, Ellen
AU - Brǎiloiu, Eugen
ID - 714
JF - Drug and Alcohol Dependence
SN - 03768716
TI - HIV Tat excites D1 receptor-like expressing neurons from rat nucleus accumbens
VL - 178
ER -
TY - JOUR
AB - D-cycloserine ameliorates breathing abnormalities and survival rate in a mouse model of Rett syndrome.
AU - Novarino, Gaia
ID - 715
IS - 405
JF - Science Translational Medicine
SN - 19466234
TI - More excitation for Rett syndrome
VL - 9
ER -
TY - JOUR
AB - Genetic variations in the oxytocin receptor gene affect patients with ASD and ADHD differently.
AU - Novarino, Gaia
ID - 731
IS - 411
JF - Science Translational Medicine
SN - 19466234
TI - The science of love in ASD and ADHD
VL - 9
ER -
TY - JOUR
AB - Since 2006, reprogrammed cells have increasingly been used as a biomedical research technique in addition to neuro-psychiatric methods. These rapidly evolving techniques allow for the generation of neuronal sub-populations, and have sparked interest not only in monogenetic neuro-psychiatric diseases, but also in poly-genetic and poly-aetiological disorders such as schizophrenia (SCZ) and bipolar disorder (BPD). This review provides a summary of 19 publications on reprogrammed adult somatic cells derived from patients with SCZ, and five publications using this technique in patients with BPD. As both disorders are complex and heterogeneous, there is a plurality of hypotheses to be tested in vitro. In SCZ, data on alterations of dopaminergic transmission in vitro are sparse, despite the great explanatory power of the so-called DA hypothesis of SCZ. Some findings correspond to perturbations of cell energy metabolism, and observations in reprogrammed cells suggest neuro-developmental alterations. Some studies also report on the efficacy of medicinal compounds to revert alterations observed in cellular models. However, due to the paucity of replication studies, no comprehensive conclusions can be drawn from studies using reprogrammed cells at the present time. In the future, findings from cell culture methods need to be integrated with clinical, epidemiological, pharmacological and imaging data in order to generate a more comprehensive picture of SCZ and BPD.
AU - Sauerzopf, Ulrich
AU - Sacco, Roberto
AU - Novarino, Gaia
AU - Niello, Marco
AU - Weidenauer, Ana
AU - Praschak Rieder, Nicole
AU - Sitte, Harald
AU - Willeit, Matthaeus
ID - 1228
IS - 1
JF - European Journal of Neuroscience
TI - Are reprogrammed cells a useful tool for studying dopamine dysfunction in psychotic disorders? A review of the current evidence
VL - 45
ER -
TY - JOUR
AB - Bradykinin (BK), a component of the kallikrein-kininogen-kinin system exerts multiple effects via B1 and B2 receptor activation. In the cardiovascular system, bradykinin has cardioprotective and vasodilator properties. We investigated the effect of BK on cardiac-projecting neurons of nucleus ambiguus, a key site for the parasympathetic cardiac regulation. BK produced a dose-dependent increase in cytosolic Ca2+ concentration. Pretreatment with HOE140, a B2 receptor antagonist, but not with R715, a B1 receptor antagonist, abolished the response to BK. A selective B2 receptor agonist, but not a B1 receptor agonist, elicited an increase in cytosolic Ca2+ similarly to BK. Inhibition of N-type voltage-gated Ca2+ channels with ω-conotoxin GVIA had no effect on the Ca2+ signal produced by BK, while pretreatment with ω-conotoxin MVIIC, a blocker of P/Q-type of Ca2+ channels, significantly diminished the effect of BK. Pretreatment with xestospongin C and 2-aminoethoxydiphenyl borate, antagonists of inositol 1,4,5-trisphosphate receptors, abolished the response to BK. Inhibition of ryanodine receptors reduced the BK-induced Ca2+ increase, while disruption of lysosomal Ca2+ stores with bafilomycin A1 did not affect the response. BK produced a dose-dependent depolarization of nucleus ambiguus neurons, which was prevented by the B2 receptor antagonist. In vivo studies indicate that microinjection of BK into nucleus ambiguus elicited bradycardia in conscious rats via B2 receptors. In summary, in cardiac vagal neurons of nucleus ambiguus, BK activates B2 receptors promoting Ca2+ influx and Ca2+ release from endoplasmic reticulum, and membrane depolarization; these effects are translated in vivo by bradycardia.
AU - Brǎiloiu, Eugen
AU - Mcguire, Matthew
AU - Shuler, Shadaria
AU - Deliu, Elena
AU - Barr, Jeffrey
AU - Abood, Mary
AU - Brailoiu, Gabriela
ID - 747
JF - Neuroscience
SN - 03064522
TI - Modulation of cardiac vagal tone by bradykinin acting on nucleus ambiguus
VL - 365
ER -
TY - JOUR
AB - Background: Long non-coding RNAs (lncRNAs) are increasingly implicated as gene regulators and may ultimately be more numerous than protein-coding genes in the human genome. Despite large numbers of reported lncRNAs, reference annotations are likely incomplete due to their lower and tighter tissue-specific expression compared to mRNAs. An unexplored factor potentially confounding lncRNA identification is inter-individual expression variability. Here, we characterize lncRNA natural expression variability in human primary granulocytes. Results: We annotate granulocyte lncRNAs and mRNAs in RNA-seq data from 10 healthy individuals, identifying multiple lncRNAs absent from reference annotations, and use this to investigate three known features (higher tissue-specificity, lower expression, and reduced splicing efficiency) of lncRNAs relative to mRNAs. Expression variability was examined in seven individuals sampled three times at 1- or more than 1-month intervals. We show that lncRNAs display significantly more inter-individual expression variability compared to mRNAs. We confirm this finding in two independent human datasets by analyzing multiple tissues from the GTEx project and lymphoblastoid cell lines from the GEUVADIS project. Using the latter dataset we also show that including more human donors into the transcriptome annotation pipeline allows identification of an increasing number of lncRNAs, but minimally affects mRNA gene number. Conclusions: A comprehensive annotation of lncRNAs is known to require an approach that is sensitive to low and tight tissue-specific expression. Here we show that increased inter-individual expression variability is an additional general lncRNA feature to consider when creating a comprehensive annotation of human lncRNAs or proposing their use as prognostic or disease markers.
AU - Kornienko, Aleksandra
AU - Dotter, Christoph
AU - Guenzl, Philipp
AU - Gisslinger, Heinz
AU - Gisslinger, Bettina
AU - Cleary, Ciara
AU - Kralovics, Robert
AU - Pauler, Florian
AU - Barlow, Denise
ID - 1240
IS - 1
JF - Genome Biology
TI - Long non-coding RNAs display higher natural expression variation than protein-coding genes in healthy humans
VL - 17
ER -
TY - JOUR
AB - Autism spectrum disorders (ASD) are a group of genetic disorders often overlapping with other neurological conditions. We previously described abnormalities in the branched-chain amino acid (BCAA) catabolic pathway as a cause of ASD. Here, we show that the solute carrier transporter 7a5 (SLC7A5), a large neutral amino acid transporter localized at the blood brain barrier (BBB), has an essential role in maintaining normal levels of brain BCAAs. In mice, deletion of Slc7a5 from the endothelial cells of the BBB leads to atypical brain amino acid profile, abnormal mRNA translation, and severe neurological abnormalities. Furthermore, we identified several patients with autistic traits and motor delay carrying deleterious homozygous mutations in the SLC7A5 gene. Finally, we demonstrate that BCAA intracerebroventricular administration ameliorates abnormal behaviors in adult mutant mice. Our data elucidate a neurological syndrome defined by SLC7A5 mutations and support an essential role for the BCAA in human brain function.
AU - Tarlungeanu, Dora-Clara
AU - Deliu, Elena
AU - Dotter, Christoph
AU - Kara, Majdi
AU - Janiesch, Philipp
AU - Scalise, Mariafrancesca
AU - Galluccio, Michele
AU - Tesulov, Mateja
AU - Morelli, Emanuela
AU - Sönmez, Fatma
AU - Bilgüvar, Kaya
AU - Ohgaki, Ryuichi
AU - Kanai, Yoshikatsu
AU - Johansen, Anide
AU - Esharif, Seham
AU - Ben Omran, Tawfeg
AU - Topcu, Meral
AU - Schlessinger, Avner
AU - Indiveri, Cesare
AU - Duncan, Kent
AU - Caglayan, Ahmet
AU - Günel, Murat
AU - Gleeson, Joseph
AU - Novarino, Gaia
ID - 1183
IS - 6
JF - Cell
TI - Impaired amino acid transport at the blood brain barrier is a cause of autism spectrum disorder
VL - 167
ER -
TY - JOUR
AB - Detecting allelic biases from high-throughput sequencing data requires an approach that maximises sensitivity while minimizing false positives. Here, we present Allelome.PRO, an automated user-friendly bioinformatics pipeline, which uses high-throughput sequencing data from reciprocal crosses of two genetically distinct mouse strains to detect allele-specific expression and chromatin modifications. Allelome.PRO extends approaches used in previous studies that exclusively analyzed imprinted expression to give a complete picture of the ‘allelome’ by automatically categorising the allelic expression of all genes in a given cell type into imprinted, strain-biased, biallelic or non-informative. Allelome.PRO offers increased sensitivity to analyze lowly expressed transcripts, together with a robust false discovery rate empirically calculated from variation in the sequencing data. We used RNA-seq data from mouse embryonic fibroblasts from F1 reciprocal crosses to determine a biologically relevant allelic ratio cutoff, and define for the first time an entire allelome. Furthermore, we show that Allelome.PRO detects differential enrichment of H3K4me3 over promoters from ChIP-seq data validating the RNA-seq results. This approach can be easily extended to analyze histone marks of active enhancers, or transcription factor binding sites and therefore provides a powerful tool to identify candidate cis regulatory elements genome wide.
AU - Andergassen, Daniel
AU - Dotter, Christoph
AU - Kulinski, Tomasz
AU - Guenzl, Philipp
AU - Bammer, Philipp
AU - Barlow, Denise
AU - Pauler, Florian
AU - Hudson, Quanah
ID - 1497
IS - 21
JF - Nucleic Acids Research
TI - Allelome.PRO, a pipeline to define allele-specific genomic features from high-throughput sequencing data
VL - 43
ER -
TY - JOUR
AB - Intellectual disability (ID) has an estimated prevalence of 2-3%. Due to its extreme heterogeneity, the genetic basis of ID remains elusive in many cases. Recently, whole exome sequencing (WES) studies revealed that a large proportion of sporadic cases are caused by de novo gene variants. To identify further genes involved in ID, we performed WES in 250 patients with unexplained ID and their unaffected parents and included exomes of 51 previously sequenced child-parents trios in the analysis. Exome analysis revealed de novo intragenic variants in SET domain-containing 5 (SETD5) in two patients. One patient carried a nonsense variant, and the other an 81 bp deletion located across a splice-donor site. Chromosomal microarray diagnostics further identified four de novo non-recurrent microdeletions encompassing SETD5. CRISPR/Cas9 mutation modelling of the two intragenic variants demonstrated nonsense-mediated decay of the resulting transcripts, pointing to a loss-of-function (LoF) and haploinsufficiency as the common disease-causing mechanism of intragenic SETD5 sequence variants and SETD5-containing microdeletions. In silico domain prediction of SETD5, a predicted SET domain-containing histone methyltransferase (HMT), substantiated the presence of a SET domain and identified a novel putative PHD domain, strengthening a functional link to well-known histone-modifying ID genes. All six patients presented with ID and certain facial dysmorphisms, suggesting that SETD5 sequence variants contribute substantially to the microdeletion 3p25.3 phenotype. The present report of two SETD5 LoF variants in 301 patients demonstrates a prevalence of 0.7% and thus SETD5 variants as a relatively frequent cause of ID.
AU - Kuechler, Alma
AU - Zink, Alexander
AU - Wieland, Thomas
AU - Lüdecke, Hermann
AU - Cremer, Kirsten
AU - Salviati, Leonardo
AU - Magini, Pamela
AU - Najafi, Kimia
AU - Zweier, Christiane
AU - Czeschik, Johanna
AU - Aretz, Stefan
AU - Endele, Sabine
AU - Tamburrino, Federica
AU - Pinato, Claudia
AU - Clementi, Maurizio
AU - Gundlach, Jasmin
AU - Maylahn, Carina
AU - Mazzanti, Laura
AU - Wohlleber, Eva
AU - Schwarzmayr, Thomas
AU - Kariminejad, Roxana
AU - Schlessinger, Avner
AU - Wieczorek, Dagmar
AU - Strom, Tim
AU - Novarino, Gaia
AU - Engels, Hartmut
ID - 1789
IS - 6
JF - European Journal of Human Genetics
TI - Loss-of-function variants of SETD5 cause intellectual disability and the core phenotype of microdeletion 3p25.3 syndrome
VL - 23
ER -
TY - JOUR
AB - Hereditary spastic paraplegias (HSPs) are neurodegenerative motor neuron diseases characterized by progressive age-dependent loss of corticospinal motor tract function. Although the genetic basis is partly understood, only a fraction of cases can receive a genetic diagnosis, and a global view of HSP is lacking. By using whole-exome sequencing in combination with network analysis, we identified 18 previously unknown putative HSP genes and validated nearly all of these genes functionally or genetically. The pathways highlighted by these mutations link HSP to cellular transport, nucleotide metabolism, and synapse and axon development. Network analysis revealed a host of further candidate genes, of which three were mutated in our cohort. Our analysis links HSP to other neurodegenerative disorders and can facilitate gene discovery and mechanistic understanding of disease.
AU - Novarino, Gaia
AU - Fenstermaker, Ali
AU - Zaki, Maha
AU - Hofree, Matan
AU - Silhavy, Jennifer
AU - Heiberg, Andrew
AU - Abdellateef, Mostafa
AU - Rosti, Başak
AU - Scott, Eric
AU - Mansour, Lobna
AU - Masri, Amira
AU - Kayserili, Hülya
AU - Al Aama, Jumana
AU - Abdel Salam, Ghada
AU - Karminejad, Ariana
AU - Kara, Majdi
AU - Kara, Bülent
AU - Bozorgmehri, Bita
AU - Ben Omran, Tawfeg
AU - Mojahedi, Faezeh
AU - Mahmoud, Iman
AU - Bouslam, Naïma
AU - Bouhouche, Ahmed
AU - Benomar, Ali
AU - Hanein, Sylvain
AU - Raymond, Laure
AU - Forlani, Sylvie
AU - Mascaro, Massimo
AU - Selim, Laila
AU - Shehata, Nabil
AU - Al Allawi, Nasir
AU - Bindu, Parayil
AU - Azam, Matloob
AU - Günel, Murat
AU - Caglayan, Ahmet
AU - Bilgüvar, Kaya
AU - Tolun, Aslihan
AU - Issa, Mahmoud
AU - Schroth, Jana
AU - Spencer, Emily
AU - Rosti, Rasim
AU - Akizu, Naiara
AU - Vaux, Keith
AU - Johansen, Anide
AU - Koh, Alice
AU - Megahed, Hisham
AU - Dürr, Alexandra
AU - Brice, Alexis
AU - Stévanin, Giovanni
AU - Gabriel, Stacy
AU - Ideker, Trey
AU - Gleeson, Joseph
ID - 1916
IS - 6170
JF - Science
TI - Exome sequencing links corticospinal motor neuron disease to common neurodegenerative disorders
VL - 343
ER -