Repurposing of tamoxifen ameliorates CLN3 and CLN7 disease phenotype

Batten diseases (BDs) are a group of lysosomal storage disorders characterized by seizure, visual loss, and cognitive and motor deterioration. We discovered increased levels of globotriaosylceramide (Gb3) in cellular and murine models of CLN3 and CLN7 diseases and used fluorescent-conjugated bacterial toxins to label Gb3 to develop a cell-based high content imaging (HCI) screening assay for the repurposing of FDA-approved compounds able to reduce this accumulation within BD cells. We found that tamoxifen reduced the lysosomal accumulation of Gb3 in CLN3 and CLN7 cell models, including neuronal progenitor cells (NPCs) from CLN7 patient-derived induced pluripotent stem cells (iPSC). Here, tamoxifen exerts its action through a mechanism that involves activation of the transcription factor EB (TFEB), a master gene of lysosomal function and autophagy. In vivo administration of tamoxifen to the CLN7Δex2 mouse model reduced the accumulation of Gb3 and SCMAS, decreased neuroinflammation, and improved motor coordination. These data strongly suggest that tamoxifen may be a suitable drug to treat some types of Batten disease

Aberrant upregulation of the glycolytic enzyme PFKFB3 in CLN7 neuronal ceroid lipofuscinosis

CLN7 neuronal ceroid lipofuscinosis is an inherited lysosomal storage neurodegenerative disease highly prevalent in children. CLN7/MFSD8 gene encodes a lysosomal membrane glycoprotein, but the biochemical processes affected by CLN7-loss of function are unexplored thus preventing development of potential treatments. Here, we found, in the Cln7∆ex2 mouse model of CLN7 disease, that failure in autophagy causes accumulation of structurally and bioenergetically impaired neuronal mitochondria. In vivo genetic approach reveals elevated mitochondrial reactive oxygen species (mROS) in Cln7∆ex2 neurons that mediates glycolytic enzyme PFKFB3 activation and contributes to CLN7 pathogenesis. Mechanistically, mROS sustains a signaling cascade leading to protein stabilization of PFKFB3, normally unstable in healthy neurons. Administration of the highly selective PFKFB3 inhibitor AZ67 in Cln7∆ex2 mouse brain in vivo and in CLN7 patients-derived cells rectifies key disease hallmarks. Thus, aberrant upregulation of the glycolytic enzyme PFKFB3 in neurons may contribute to CLN7 pathogenesis and targeting PFKFB3 could alleviate this and other lysosomal storage diseases.This work was funded by the European Regional Development Fund, European Union’s Horizon 2020 Research and Innovation Programme (BATCure grant No. 666918 to J.P.B., S.E.M., D.L.M., S.S., and T.R.M.; PANA grant No. 686009 to A.A.), Agencia Estatal de Investigación (PID2019-105699RB-I00/AEI/10.13039/501100011033 and RED2018‐102576‐T to J.P.B.; SAF2017-90794-REDT to A.A.), Instituto de Salud Carlos III (CB16/10/00282 to J.P.B.; PI18/00285; RD16/0019/0018 to A.A.), Junta de Castilla y León (CS/151P20 and Escalera de Excelencia CLU-2017-03 to J.P.B. and A.A.), Ayudas Equipos Investigación Biomedicina 2017 Fundación BBVA (to J.P.B.), and Fundación Ramón Areces (to J.P.B. and A.A.). SM benefits from MRC funding to the MRC Laboratory for Molecular Cell Biology University Unit at UCL (award code MC_U12266B) towards lab and office space. Part of this work was funded by Gero Discovery L.L.C. M.G.M. is an ISCIII-Sara Borrel contract recipient (CD18/00203)

A high throughput genotyping approach reveals distinctive autosomal genetic signatures for European and Near Eastern wild boar

The lack of a Near Eastern genetic signature in modern European porcine breeds indicates that, although domestic pigs from the Fertile Crescent entered Europe during the Neolithic, they were completely replaced by their European counterparts in a short window of time. Whilst the absence of such genetic signature has been convincingly demonstrated at the mitochondrial level, variation at the autosomal genomes of European and Near Eastern Sus scrofa has not been compared yet. Herewith, we have explored the genetic relationships among 43 wild boar from Europe (N = 21), Near East (N = 19) and Korea (N = 3), and 40 Iberian (N = 16), Canarian (N = 4) and Mangalitza (N = 20) pigs by using a high throughput SNP genotyping platform. After data filtering, 37,167 autosomal SNPs were used to perform population genetics analyses. A multidimensional scaling plot based on genome-wide identity-by-state pairwise distances inferred with PLINK showed that Near Eastern and European wild boar populations are genetically differentiated. Maximum likelihood trees built with TreeMix supported this conclusion i.e. an early population split between Near Eastern and European Sus scrofa was observed. Moreover, analysis of the data with Structure evidenced that the sampled Iberian, Canarian and Mangalitza pigs did not carry any autosomal signature compatible with a Near Eastern ancestry, a finding that agrees well with previous mitochondrial studies

Analysis of conservation priorities of Iberoamerican cattle based on autosomal microsatellite markers

Articles in International JournalsBackground: Determining the value of livestock breeds is essential to define conservation priorities, manage genetic diversity and allocate funds. Within- and between-breed genetic diversity need to be assessed to preserve the highest intra-specific variability. Information on genetic diversity and risk status is still lacking for many Creole cattle breeds from the Americas, despite their distinct evolutionary trajectories and adaptation to extreme environmental conditions. Methods: A comprehensive genetic analysis of 67 Iberoamerican cattle breeds was carried out with 19 FAOrecommended microsatellites to assess conservation priorities. Contributions to global diversity were investigated using alternative methods, with different weights given to the within- and between-breed components of genetic diversity. Information on Iberoamerican plus 15 worldwide cattle breeds was used to investigate the contribution of geographical breed groups to global genetic diversity. Results: Overall, Creole cattle breeds showed a high level of genetic diversity with the highest level found in breeds admixed with zebu cattle, which were clearly differentiated from all other breeds. Within-breed kinships revealed seven highly inbred Creole breeds for which measures are needed to avoid further genetic erosion. However, if contribution to heterozygosity was the only criterion considered, some of these breeds had the lowest priority for conservation decisions. The Weitzman approach prioritized highly differentiated breeds, such as Guabalá, Romosinuano, Cr. Patagonico, Siboney and Caracú, while kinship-based methods prioritized mainly zebu-related breeds. With the combined approaches, breed ranking depended on the weights given to the within- and between-breed components of diversity. Overall, the Creole groups of breeds were generally assigned a higher priority for conservation than the European groups of breeds. Conclusions: Conservation priorities differed significantly according to the weight given to within- and betweenbreed genetic diversity. Thus, when establishing conservation programs, it is necessary to also take into account other features. Creole cattle and local isolated breeds retain a high level of genetic diversity. The development of sustainable breeding and crossbreeding programs for Creole breeds, and the added value resulting from their products should be taken into consideration to ensure their long-term survival

Anti-tumour necrosis factor discontinuation in inflammatory bowel disease patients in remission: study protocol of a prospective, multicentre, randomized clinical trial

Background: Patients with inflammatory bowel disease who achieve remission with anti-tumour necrosis factor (anti-TNF) drugs may have treatment withdrawn due to safety concerns and cost considerations, but there is a lack of prospective, controlled data investigating this strategy. The primary study aim is to compare the rates of clinical remission at 1?year in patients who discontinue anti-TNF treatment versus those who continue treatment. Methods: This is an ongoing, prospective, double-blind, multicentre, randomized, placebo-controlled study in patients with Crohn?s disease or ulcerative colitis who have achieved clinical remission for ?6?months with an anti-TNF treatment and an immunosuppressant. Patients are being randomized 1:1 to discontinue anti-TNF therapy or continue therapy. Randomization stratifies patients by the type of inflammatory bowel disease and drug (infliximab versus adalimumab) at study inclusion. The primary endpoint of the study is sustained clinical remission at 1?year. Other endpoints include endoscopic and radiological activity, patient-reported outcomes (quality of life, work productivity), safety and predictive factors for relapse. The required sample size is 194 patients. In addition to the main analysis (discontinuation versus continuation), subanalyses will include stratification by type of inflammatory bowel disease, phenotype and previous treatment. Biological samples will be obtained to identify factors predictive of relapse after treatment withdrawal. Results: Enrolment began in 2016, and the study is expected to end in 2020. Conclusions: This study will contribute prospective, controlled data on outcomes and predictors of relapse in patients with inflammatory bowel disease after withdrawal of anti-TNF agents following achievement of clinical remission. Clinical trial reference number: EudraCT 2015-001410-1

Alternativas metodológicas para el cálculo de las series retrospectivas regionales de EPA tras el cambio en la Clasificación Nacional de Actividades Económicas // Alternative Approaches for Calculating the Retrospective Regional EPA Series Following the Changes in the National Classification of Economic Activities

A fin de solventar la potencial pérdida de información que la entrada en vigor de la nueva Clasificación Nacional de Actividades Económicas provocaría en las series de EPA, el INE ha propuesto una metodología que permite el enlace de las series CNAE 1993 y CNAE 2009. El procedimiento de enlace se basa en trabajar con una matriz de paso nacional sobre los datos a nivel de división. Los mercados de trabajo regionales, sin embargo, difieren del nacional y, además, las series regionales no suelen estar disponibles a nivel de división. Este trabajo estudia qué impacto tendría en las series enlazadas (i) trabajar en un nivel de agregación superior al de división y (ii) utilizar una matriz de paso regional propia. Centrada en las series de ocupados publicadas para la Comunidad Valenciana, la investigación muestra que: (a) a nivel sectorial, es equivalente emplear la matriz de España o una propia y, también, el trasladar las divisiones y luego agregarlas que el trasladar directamente las series agregadas; y (b) que a nivel de rama de actividad, es preferible trabajar con una matriz propia.------------------------------------In order to overcome the potential loss of information that the transition to the new National Classification of Economic Activities could provoke in EPA series, INE has proposed a methodology to link CNAE 1993 and CNAE 2009 series. The procedure is based on employing a national transition matrix over division data. Regional labor markets, however, differ from the national one and additionally regional series are not generally available at the division level. This paper examines the impact that in the linked series should have (i) working at a level of aggregation above division and (ii) using a regional transition matrix. Focusing employment series published for the Valencian region, this research shows that (a), at the sectorial level, results do not depend on use of either the Spanish transition matrix of Spain or the Valencian one and also they are independent of the order in which aggregation of the divisions takes place and that (b), at the branch of activity, it is preferable to work with an own regional matrix

Alternativas metodológicas para el cálculo de las series retrospectivas regionales de EPA tras el cambio en la Clasificación Nacional de Actividades Económicas || Alternative Approaches for Calculating the Retrospective Regional EPA Series Following the Changes in the National Classification of Economic Activities

A fin de solventar la potencial pérdida de información que la entrada en vigor de la nueva Clasificación Nacional de Actividades Económicas provocaría en las series de EPA, el INE ha propuesto una metodología que permite el enlace de las series CNAE 1993 y CNAE 2009. El procedimiento de enlace se basa en trabajar con una matriz de paso nacional sobre los datos a nivel de división. Los mercados de trabajo regionales, sin embargo, difieren del nacional y, además, las series regionales no suelen estar disponibles a nivel de división. Este trabajo estudia qué impacto tendría en las series enlazadas (i) trabajar en un nivel de agregación superior al de división y (ii) utilizar una matriz de paso regional propia. Centrada en las series de ocupados publicadas para la Comunidad Valenciana, la investigación muestra que: (a) a nivel sectorial, es equivalente emplear la matriz de España o una propia y, también, el trasladar las divisiones y luego agregarlas que el trasladar directamente las series agregadas; y (b) que a nivel de rama de actividad, es preferible trabajar con una matriz propia. || In order to overcome the potential loss of information that the transition to the new National Classification of Economic Activities could provoke in EPA series, INE has proposed a methodology to link CNAE 1993 and CNAE 2009 series. The procedure is based on employing a national transition matrix over division data. Regional labour markets, however, differ from the national one and additionally regional series are not generally available at the division level. This paper examines the impact that in the linked series should have (i) working at a level of aggregation above division and (ii) using a regional transition matrix. Focusing employment series published for the Valencian region, this research shows that (a), at the sectorial level, results do not depend on use of either the Spanish transition matrix of Spain or the Valencian one and also they are independent of the order in which aggregation of the divisions takes place and that (b), at the branch of activity, it is preferable to work with an own regional matrix

Alternativas metodológicas para el cálculo de las series retrospectivas regionales de EPA tras el cambio en la Clasificación Nacional de Actividades Económicas

In order to overcome the potential loss of information that the transition to the new National Classification of Economic Activities could provoke in EPA series, INE has proposed a methodology to link CNAE 1993 and CNAE 2009 series. The procedure is based on employing a national transition matrix over division data. Regional labor markets, however, differ from the national one and additionally regional series are not generally available at the division level. This paper examines the impact that in the linked series should have (i) working at a level of aggregation above division and (ii) using a regional transition matrix. Focusing employment series published for the Valencian region, this research shows that (a), at the sectorial level, results do not depend on use of either the Spanish transition matrix of Spain or the Valencian one and also they are independent of the order in which aggregation of the divisions takes place and that (b), at the branch of activity, it is preferable to work with an own regional matrix

Complex I assembly into supercomplexes determines differential mitochondrial ROS production in neurons and astrocytes

Neurons depend on oxidative phosphorylation for survival, whereas astrocytes do not. Mitochondrial respiratory chain (MRC) complexes can be organized in higher structures called supercomplexes, which dictate MRC electron flux and energy efficiency. Whether the specific metabolic shapes of neurons and astrocytes are determined by the specific organization of MRC complexes is unknown. Here, we found that, in astrocytes, most complex I is free, resulting in poor mitochondrial respiration but high reactive oxygen species (ROS) production. In contrast, neurons show complex I to be mostly embedded into supercomplexes, thus resulting in high mitochondrial respiration and low ROS production. Thus, MRC organization dictates different bioenergetics preferences of neurons and astrocytes impacting on ROS production, possibly playing a role in neurodegenerative diseases.J.P.B. is funded by the Ministry of Economy and Competitiveness (SAF2013-41177-R), Instituto de Salud Carlos III (RD12/0043/0021), European Union (EU) SP3-People-MC-ITN Programme (608381), EU BATCure Grant 666918, and NIH/National Institute on Drug Abuse Grant 1R21DA037678-01. A.A. is funded by Instituto de Salud Carlos III (PI12/00685 and RD12/0014/0007).Peer Reviewe