ALTERACIONES PROTEOSTATICAS TEMPRANAS EN LAS ENFERMEDADES DE ALZHEIMER Y PARKINSON A NIVEL OLFATORIO: PAPEL DE GNPDA2 Y PITHD1 DURANTE EL PROCESO DE NEURODEGENERACION

PID2019-110356RB-I00

Nombre agencia financiadora Agencia Estatal de Investigación
Acrónimo agencia financiadora AEI
Programa Programa Estatal de Generación de Conocimiento y Fortalecimiento Científico y Tecnológico del Sistema de I+D+i
Subprograma Subprograma Estatal de Generación de Conocimiento
Convocatoria Proyectos I+D
Año convocatoria 2019
Unidad de gestión Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020
Centro beneficiario FUNDACION MIGUEL SERVET
Identificador persistente http://dx.doi.org/10.13039/501100011033

Publicaciones

Found(s) 8 result(s)
Found(s) 1 page(s)

Brain IGF-I is essential for LTP, and it regulates sexual dimorphic behaviour and proteostasis

Digital.CSIC. Repositorio Institucional del CSIC
  • Herrero-Labrador, Raquel
  • Fernández-Irigoyen, Joaquín
  • Vecino, Rebeca
  • González-Arias, Candela
  • Ausin, K.
  • Crespo, Inmaculada
  • Fernández Acosta, F.J.
  • Nieto-Estévez, Vanesa
  • Roman, María José
  • Perea, Gertrudis
  • Torres-Alemán, Ignacio
  • Santamaria, E.
  • Vicario-Abejón, Carlos
Insulin-like growth factor-I (IGF-I) exerts multiple actions, yet the role of IGF-I from different sources is poorly understood. Here, we explored the functional and behavioral consequences of the conditional deletion of Igf-I in the nervous system (Igf-IΔ/Δ), and demonstrated that long-term potentiation was impaired in hippocampal slices. Moreover, Igf-IΔ/Δ mice showed spatial memory deficits in the Morris water maze, and the significant sex-dependent differences displayed by Igf-ICtrl/Ctrl mice disappeared in Igf-IΔ/Δ mice in the open field and rota-rod tests. Brain Igf-I deletion disorganized the granule cell layer of the dentate gyrus (DG), and it modified the relative expressions of GAD and VGLUT1, which are preferentially localized to inhibitory and excitatory presynaptic terminals. Furthermore, Igf-I deletion altered protein modules involved in receptor trafficking, synaptic proteins, and proteins that functionally interact with estrogen and androgen metabolism. Our findings indicate that brain IGF-I is crucial for long-term potentiation, and that it is involved in the regulation of spatial memory and sexual dimorphic behaviors, possibly by maintaining the granule cell layer structure and the stability of synaptic-related protein modules., We thank Lucía Vicario (Instituto Cajal-CSIC, Madrid, Spain) for helping with the composition of the figures, and Dr. M Sefton (BiomedRed SL, Madrid, Spain) for English editing. This work was funded by grants from the Spanish “Ministerio Ciencia, Innovación y Universidades, and the Ministerio de Ciencia e Innovación/Agencia Estatal de Investigación” (MICIU and MICINN/AEI SAF2016-80419-R, PID2019-109059RB-100, and CIBERNED CB06/05/0065 to C Vicario; PID2019-110356RB-I00/AEI/10.13039/501100011033 to J Fernández-Irigoyen and E Santamaría; PID2019-104376RB-I00 to I Torres-Alemán; and PID2019-106579RB-I00 funded by MCIN/AEI/10.13039/501100011033 and by “ERDF A way of making Europe to G Perea, and BES-2017-080303 to C González-Arias).




Smelling the dark proteome: Functional characterization of PITH domain-containing protein 1 (C1orf128) in olfactory metabolism

Dadun. Depósito Académico Digital de la Universidad de Navarra
  • Lachén-Montes, M. (Mercedes)
  • Mendizuri, N. (Naroa)
  • Ausin, K. (Karina)
  • Perez-Mediavilla, L.A. (Luis Alberto)
  • Azkargorta, M. (Mikel)
  • Iloro, I. (Ibon)
  • Elortza, F. (Felix)
  • Kondo, H. (Hiroyuki)
  • Ohigashi, I. (Izumi)
  • Ferrer, I. (Isidro)
  • Torre, R. (Rafael) de la
  • Robledo-Ramón, P. (Patricia)
  • Fernandez-Irigoyen, J. (Joaquín)
  • Santamaria, E. (Enrique)
The Human Proteome Project (HPP) consortium aims to functionally characterize the dark proteome. On the basis of the relevance of olfaction in early neurodegeneration, we have analyzed the dark proteome using data mining in public resources and omics data sets derived from the human olfactory system. Multiple dark proteins localize at synaptic terminals and may be involved in amyloidopathies such as Alzheimer’s disease (AD). We have characterized the dark PITH domain-containing protein 1 (PITHD1) in olfactory metabolism using bioinformatics, proteomics, in vitro and in vivo studies, and neuropathology. PITHD1–/– mice exhibit olfactory bulb (OB) proteome changes related to synaptic transmission, cognition, and memory. OB PITHD1 expression increases with age in wild-type (WT) mice and decreases in Tg2576 AD mice at late stages. The analysis across 6 neurological disorders reveals that olfactory tract (OT) PITHD1 is specifically upregulated in human AD. Stimulation of olfactory neuroepithelial (ON) cells with PITHD1 alters the ON phosphoproteome, modifies the proliferation rate, and induces a pro-inflammatory phenotype. This workflow applied by the Spanish C-HPP and Human Brain Proteome Project (HBPP) teams across the ON-OB-OT axis can be adapted as a guidance to decipher functional features of dark proteins. Data are available via ProteomeXchange with identifiers PXD018784 and PXD021634.




Cannabis Use Induces Distinctive Proteomic Alterations in Olfactory Neuroepithelial Cells of Schizophrenia Patients

Dipòsit Digital de Documents de la UAB
  • Barrera-Conde, Marta|||0000-0002-0106-0000
  • Ausin, Karina|||0000-0003-4914-9466
  • Lachén-Montes, Mercedes
  • Fernández-Irigoyen, Joaquín|||0000-0001-5072-4099
  • Galindo, Liliana|||0000-0002-2396-4772
  • Cuenca-Royo, Aida|||0000-0001-8551-5457
  • Fernández-Avilés, Cristina|||0000-0002-1029-9216
  • Pérez Solà, Víctor|||0000-0002-5825-2337
  • De La Torre, Rafael|||0000-0002-6765-1866
  • Santamaría, Enrique
  • Robledo, Patricia
A close epidemiological link has been reported between cannabis use and schizophrenia (SCZ). However, biochemical markers in living humans related to the impact of cannabis in this disease are still missing. Olfactory neuroepithelium (ON) cells express neural features and offer a unique advantage to study biomarkers of psychiatric diseases. The aim of our study was to find exclusively deregulated proteins in ON cells of SCZ patients with and without a history of cannabis use. Thus, we compared the proteomic profiles of SCZ non-cannabis users (SCZ/nc) and SCZ cannabis users (SCZ/c) with control subjects non-cannabis users (C/nc) and control cannabis users (C/c). The results revealed that the main cascades affected in SCZ/nc were cell cycle, DNA replication, signal transduction and protein localization. Conversely, cannabis use in SCZ patients induced specific alterations in metabolism of RNA and metabolism of proteins. The levels of targeted proteins in each population were then correlated with cognitive performance and clinical scores. In SCZ/c, the expression levels of 2 proteins involved in the metabolism of RNA (MTREX and ZNF326) correlated with several cognitive markers and clinical signs. Moreover, use duration of cannabis negatively correlated with ZNF326 expression. These findings indicate that RNA-related proteins might be relevant to understand the influence of cannabis use on SCZ.




Smelling the dark proteome: functional characterization of PITH domain-containing protein 1 (C1orf128) in olfactory metabolism

Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
  • 0000-0003-2449-8117
  • Mendizuri, Naroa
  • Ausín, Karina
  • Pérez Mediavilla, Alberto
  • Azkargorta, Mikel
  • 0000-0001-5072-4099
  • Santamaría, Enrique
  • Iloro, Ibon
  • Elortza, Félix
  • Kondo, Hiroyuki
  • Ohigashi, Izumi
  • Ferrer, Isidro
  • Torre, Rafael de la
  • Robledo, Patricia
Incluye material complementario, The Human Proteome Project (HPP) consortium aims to functionally characterize the dark proteome. On the basis of the relevance of olfaction in early neurodegeneration, we have analyzed the dark proteome using data mining in public resources and omics data sets derived from the human olfactory system. Multiple dark proteins localize at synaptic terminals and may be involved in amyloidopathies such as Alzheimer's disease (AD). We have characterized the dark PITH domain-containing protein 1 (PITHD1) in olfactory metabolism using bioinformatics, proteomics, in vitro and in vivo studies, and neuropathology. PITHD1-/- mice exhibit olfactory bulb (OB) proteome changes related to synaptic transmission, cognition, and memory. OB PITHD1 expression increases with age in wild-type (WT) mice and decreases in Tg2576 AD mice at late stages. The analysis across 6 neurological disorders reveals that olfactory tract (OT) PITHD1 is specifically upregulated in human AD. Stimulation of olfactory neuroepithelial (ON) cells with PITHD1 alters the ON phosphoproteome, modifies the proliferation rate, and induces a pro-inflammatory phenotype. This workflow applied by the Spanish C-HPP and Human Brain Proteome Project (HBPP) teams across the ON-OB-OT axis can be adapted as a guidance to decipher functional features of dark proteins. Data are available via ProteomeXchange with identifiers PXD018784 and PXD021634., This work was funded by grants from the Spanish Ministry of Science Innovation and Universities (ref. PID2019-110356RB-I00 to JFI and ES) and Department of Economic and Business Development from Government of Navarra (ref. 0011-1411-2020-000028) to ES and from MEXT-JSPS 17K08884 and Takeda Science Foundation to IO. The Proteomics Unit of Navarrabiomed is a member of Proteored (PRB3-ISCIII) and is supported by grant PT17/0019/009 to JFI, of the PE I+D+I 2013–2016 funded by ISCIII and FEDER. MLM was supported by a postdoctoral fellowship from the Public University of Navarra (UPNA).




Increased C-X-C motif chemokine ligand 12 levels in cerebrospinal fluid as a candidate biomarker in sporadic amyotrophic lateral sclerosis

Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
  • Andrés Benito, Pol
  • Povedano, Mónica
  • Domínguez Rubio, Raúl
  • Marco, Carla
  • Colomina, María J.
  • López-Pérez, Óscar
  • Santana, Isabel
  • Baldeiras, Inês
  • Martínez-Yelámos, Sergio
  • Zerr, Inga
  • Llorens, Franc
  • 0000-0001-5072-4099
  • Santamaría, Enrique
  • Ferrer, Isidro
Sporadic amyotrophic lateral sclerosis (sALS) is a fatal progressive neurodegenerative disease affecting upper and lower motor neurons. Biomarkers are useful to facilitate the diagnosis and/or prognosis of patients and to reveal possible mechanistic clues about the disease. This study aimed to identify and validate selected putative biomarkers in the cerebrospinal fluid (CSF) of sALS patients at early disease stages compared with age-matched controls and with other neurodegenerative diseases including Alzheimer disease (AD), spinal muscular atrophy type III (SMA), frontotemporal dementia behavioral variant (FTD), and multiple sclerosis (MS). SWATH acquisition on liquid chromatography-tandem mass spectrometry (LC–MS/MS) for protein quantitation, and ELISA for validation, were used in CSF samples of sALS cases at early stages of the disease. Analysis of mRNA and protein expression was carried out in the anterior horn of the lumbar spinal cord in post-mortem tissue of sALS cases (terminal stage) and controls using RTq-PCR, and Western blotting, and immunohistochemistry, respectively. SWATH acquisition on liquid chromatography-tandem mass spectrometry (LC–MS/MS) revealed 51 differentially expressed proteins in the CSF in sALS. Receiver operating characteristic (ROC) curves showed CXCL12 to be the most valuable candidate biomarker. We validated the values of CXCL12 in CSF with ELISA in two different cohorts. Besides sALS, increased CXCL12 levels were found in MS but were not altered in AD, SMA, and FTD. Therefore, increased CXCL12 levels in the CSF can be useful in the diagnoses of MS and sALS in the context of the clinical settings. CXCL12 immunoreactivity was localized in motor neurons in control and sALS, and in a few glial cells in sALS at the terminal stage; CXCR4 was in a subset of oligodendroglial-like cells and axonal ballooning of motor neurons in sALS; and CXCR7 in motor neurons in control and sALS, and reactive astrocytes in the pyramidal tracts in terminal sALS. CXCL12/CXCR4/CXCR7 axis in the spinal cord probably plays a complex role in inflammation, oligodendroglial and astrocyte signaling, and neuronal and axonal preservation in sALS., The project leading to these results received funding from Fundació Miquel Valls, 'Retos todos unidos contra la ELA' and 'Proyecto DGeneracion conexiones con sentido' to MP. The study was also supported by the Ministry of Economy and Competitiveness, Institute of Health Carlos III (ISCIII) (co-funded by European RegiSonal Development Fund, ERDF, a way to build Europe): FISPI17/000809 to IF; and FIS (ISCIII) grant PI19/00144 to FLl. The Proteomics Unit of Navarrabiomed is supported by grant PT17/0019/009 to JFI, of the PE I+D+I 2013-2016 funded by ISCIII and FEDER. Part of this work was funded by a grant from the Spanish Ministry of Science Innovation and Universities (Ref. PID2019-110356RB-I00) to JFI and ES.




Metabolic dyshomeostasis induced by SARS-CoV-2 structural proteins reveals immunological insights into viral olfactory interactions

Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
  • Lachén Montes, Mercedes
  • Mendizuri, Naroa
  • Ausín, Karina
  • Echaide Górriz, Míriam
  • Blanco, Ester
  • 0000-0001-7384-9847
  • Toro, María de
  • Escors Murugarren, David
  • 0000-0001-5072-4099
  • Kochan, Grazyna
  • Santamaría, Enrique
One of the most common symptoms in COVID-19 is a sudden loss of smell. SARS-CoV-2 has been detected in the olfactory bulb (OB) from animal models and sporadically in COVID-19 patients. To decipher the specific role over the SARS-CoV-2 proteome at olfactory level, we characterized the in-depth molecular imbalance induced by the expression of GFP-tagged SARS-CoV-2 structural proteins (M, N, E, S) on mouse OB cells. Transcriptomic and proteomic trajectories uncovered a widespread metabolic remodeling commonly converging in extracellular matrix organization, lipid metabolism and signaling by receptor tyrosine kinases. The molecular singularities and specific interactome expression modules were also characterized for each viral structural factor. The intracellular molecular imbalance induced by each SARS-CoV-2 structural protein was accompanied by differential activation dynamics in survival and immunological routes in parallel with a differentiated secretion profile of chemokines in OB cells. Machine learning through a proteotranscriptomic data integration uncovered TGF-beta signaling as a confluent activation node by the SARS-CoV-2 structural proteome. Taken together, these data provide important avenues for understanding the multifunctional immunomodulatory properties of SARS-CoV-2 M, N, S and E proteins beyond their intrinsic role in virion formation, deciphering mechanistic clues to the olfactory inflammation observed in COVID-19 patients., This work was funded by grants from the Spanish Ministry of Science, Innovation and Universities (Ref. PID2019-110356RB-I00/AEI/10.13039/501100011033) to JF-I. and ES), the Department of Economic and Business Development from Government of Navarra (Ref. 0011-1411-2020-000028 to ES), the Instituto de Salud Carlos III (ISCIII)-FEDER project grants (Ref. FIS PI17/02119, FIS PI20/00010; COV20-00237 to DE), the Department of Health of the Government of Navarre (Ref: BMED 050-2019 to DE) and the European Project Horizon 2020 (ref: ID: 848166; Improved vaccination for older adults-ISOLDA to DE).




Deregulated transcription and proteostasis in adult mapt knockout mouse

Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
  • Andrés Benito, Pol
  • Flores, África
  • Busquet-Areny, Sara
  • Carmona, Margarita
  • Ausín, Karina
  • Cartas Cejudo, Paz
  • 0000-0003-2449-8117
  • Río, José Antonio del
  • 0000-0001-5072-4099
  • Santamaría, Enrique
  • Ferrer, Isidro
Transcriptomics and phosphoproteomics were carried out in the cerebral cortex of B6.Cg-Mapttm1(EGFP)Klt (tau knockout: tau-KO) and wild-type (WT) 12 month-old mice to learn about the effects of tau ablation. Compared with WT mice, tau-KO mice displayed reduced anxiety-like behavior and lower fear expression induced by aversive conditioning, whereas recognition memory remained unaltered. Cortical transcriptomic analysis revealed 69 downregulated and 105 upregulated genes in tau-KO mice, corresponding to synaptic structures, neuron cytoskeleton and transport, and extracellular matrix components. RT-qPCR validated increased mRNA levels of col6a4, gabrq, gad1, grm5, grip2, map2, rab8a, tubb3, wnt16, and an absence of map1a in tau-KO mice compared with WT mice. A few proteins were assessed with Western blotting to compare mRNA expression with corresponding protein levels. Map1a mRNA and protein levels decreased. However, ¿-tubulin III and GAD1 protein levels were reduced in tau-KO mice. Cortical phosphoproteomics revealed 121 hypophosphorylated and 98 hyperphosphorylated proteins in tau-KO mice. Deregulated phosphoproteins were categorized into cytoskeletal (n = 45) and membrane proteins, including proteins of the synapses and vesicles, myelin proteins, and proteins linked to membrane transport and ion channels (n = 84), proteins related to DNA and RNA metabolism (n = 36), proteins connected to the ubiquitin-proteasome system (UPS) (n = 7), proteins with kinase or phosphatase activity (n = 21), and 22 other proteins related to variegated pathways such as metabolic pathways, growth factors, or mitochondrial function or structure. The present observations reveal a complex altered brain transcriptome and phosphoproteome in tau-KO mice with only mild behavioral alterations., The project leading to these results received funding from the “la Caixa” Foundation (ID 100010434) under the agreements LCF/PR/HR19/52160007, HR18-00452 to IF. We thank the CERCA Programme/Generalitat de Catalunya for institutional support. The Proteomics Platform of Navarrabiomed is a member of Proteored (PRB3-ISCIII), supported by grant PT17/0019/009 to J.F.-I. of the PE I + D + I 2013-2016 funded by ISCIII and FEDER. This work was funded by a grant from the Spanish Ministry of Science, Innovation, and Universities (Ref. PID2019-110356RB-I00) to J.F.-I. and E.S. and the Department of Economic and Business Development from the Government of Navarra (Ref. 0011-1411-2020-000028) to E.S.




Proteostatic modulation in brain aging without associated Alzheimer's disease-and age-related neuropathological changes

Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
  • Andrés Benito, Pol
  • Íñigo-Marco, Ignacio
  • Brullas, Marta
  • Carmona, Margarita
  • Del Rio, José Antonio
  • 0000-0001-5072-4099
  • Santamaría, Enrique
  • Povedano, Mónica
  • Ferrer, Isidro
Aims: (Phospho)proteomics of old-aged subjects without cognitive or behavioral symptoms, and without ADneuropathological changes and lacking any other neurodegenerative alteration will increase understanding about
the physiological state of human brain aging without associate neurological deficits and neuropathological lesions.
Methods: (Phospho)proteomics using conventional label-free- and SWATH-MS (Sequential window acquisition of
all theoretical fragment ion spectra mass spectrometry) has been assessed in the frontal cortex (FC) of individuals
without NFTs, senile plaques (SPs) and age-related co-morbidities classified by age (years) in four groups; group 1
(young, 30–44); group 2 (middle-aged: MA, 45-52); group 3 (early-elderly, 64–70); and group 4 (late-elderly, 75–85).
Results: Protein levels and deregulated protein phosphorylation linked to similar biological terms/functions,
but involving different individual proteins, are found in FC with age. The modified expression occurs in
cytoskeleton proteins, membranes, synapses, vesicles, myelin, membrane transport and ion channels, DNA and
RNA metabolism, ubiquitin-proteasome-system (UPS), kinases and phosphatases, fatty acid metabolism, and
mitochondria. Dysregulated phosphoproteins are associated with the cytoskeleton, including microfilaments,
actin-binding proteins, intermediate filaments of neurons and glial cells, and microtubules; membrane proteins,
synapses, and dense core vesicles; kinases and phosphatases; proteins linked to DNA and RNA; members of the
UPS; GTPase regulation; inflammation; and lipid metabolism. Noteworthy, protein levels of large clusters of
hierarchically-related protein expression levels are stable until 70. However, protein levels of components of cell membranes, vesicles and synapses, RNA modulation, and cellular structures (including tau and tubulin
filaments) are markedly altered from the age of 75. Similarly, marked modifications occur in the larger
phosphoprotein clusters involving cytoskeleton and neuronal structures, membrane stabilization, and kinase
regulation in the late elderly.
Conclusions: Present findings may increase understanding of human brain proteostasis modifications in the
elderly in the subpopulation of individuals not having AD neuropathological change and any other
neurodegenerative change in any telencephalon region., The project leading to these results received funding from
the “la Caixa” Foundation (ID 100010434) under the
agreement LCF/PR/HR19/52160007, HR18-00452 to IF
and JAdR. We thank CERCA Programme/Generalitat de
Catalunya for institutional support. The Proteomics
Platform of Navarrabiomed is a member of Proteored
(PRB3-ISCIII) and is supported by grant PT17/0019/009
to JFI, of the PE I+D+I 2013–2016 funded by ISCIII and
FEDER. Parts of this work were funded by a grant from
the Spanish Ministry of Science Innovation and
Universities (Ref. PID2019-110356RB-I00) to JFI and
ES, and the Department of Economic and Business Development of the Government of Navarra (Ref. 0011-
1411-2020-000028) to ES.