Intestinal microbiota modulation in obesity-related non-alcoholic fatty liver disease

[EN] Obesity and associated comorbidities, including non-alcoholic fatty liver disease (NAFLD), are a major concern to public well-being worldwide due to their high prevalence among the population, and its tendency on the rise point to as important threats in the future. Therapeutic approaches for obesity-associated disorders have been circumscribed to lifestyle modifications and pharmacological therapies have demonstrated limited efficacy. Over the last few years, different studies have shown a significant role of intestinal microbiota (IM) on obesity establishment and NAFLD development. Therefore, modulation of IM emerges as a promising therapeutic strategy for obesity-associated diseases. Administration of prebiotic and probiotic compounds, fecal microbiota transplantation (FMT) and exercise protocols have shown a modulatory action over the IM. In this review we provide an overview of current approaches targeting IM which have shown their capacity to counteract NAFLD and metabolic syndrome features in human patients and animal models.SIThis work was supported by grants to JG-G and SS-C from Ministerio de Economía y Competitividad/FEDER (BFU2017- 87960-R) and Junta de Castilla y León (LE063U16 and GRS 1888/A/18). DP was supported by a fellowship from Junta de Castilla y León co-financed by the European Social Fund. EN was supported by Fundación de Investigación Sanitaria of León. MG-M was supported by CIBERehd contracts. CIBERehd is funded by the Instituto de Salud Carlos III, Spain

Functional Interactions between Gut Microbiota Transplantation, Quercetin, and High‐Fat Diet Determine Non‐Alcoholic Fatty Liver Disease Development in Germ‐Free Mice

[EN] Scope: Modulation of intestinal microbiota has emerged as a new therapeutic approach for non-alcoholic fatty liver disease (NAFLD). Herein, it is addressed whether gut microbiota modulation by quercetin and intestinal microbiota transplantation can influence NAFLD development. Methods and results: Gut microbiota donor mice are selected according to their response to high-fat diet (HFD) and quercetin in terms of obesity and NAFLD-related biomarkers. Germ-free recipients displayed metabolic phenotypic differences derived from interactions between microbiota transplanted, diets, and quercetin. Based on the evaluation of hallmark characteristics of NAFLD, it is found that gut microbiota transplantation from the HFD-non-responder donor and the HFD-fed donor with the highest response to quercetin results in a protective phenotype against HFD-induced NAFLD, in a mechanism that involves gut–liver axis alteration blockage in these receivers. Gut microbiota from the HFD-responder donor predisposed transplanted germ-free mice to NAFLD. Divergent protective and deleterious metabolic phenotypes exhibited are related to definite microbial profiles in recipients, highlighting the predominant role of Akkermansia genus in the protection from obesity-associated NAFLD development. Conclusions: The results provide scientific support for the prebiotic capacity of quercetin and the transfer of established metabolic profiles through gut microbiota transplantation as a protective strategy against the development of obesity-related NAFLDSIM.V.G.M. and S.S.C. share senior authorship. D.P. and E.N. made equal contribution to the study. D.P., E.N., S.M.F., M.V.G.M., and S.S.C. performed most of the experiments. J.L.O. and F.J. performed statistical analysis. R.J. and J.G.G. assisted for in vivo models. S.S.C. designed the experiments and supervised the study. All the authors wrote the manuscript. The authors thank Drs. Gérard and Rabot, from MICALIS Institute (INRA), for providing germ-free mice. This work was supported by grants from Ministerio de Economía y Competitividad and Fondo Europeo de Desarrollo Regional (FEDER) (BFU2013-48141-R, BFU2017-87960-R), Junta de Castilla y León (LE135U13, GRS 1428/A/16), Junta de Castilla y León and FEDER, (LE063U16), and IIS Hospital La Fe (2017_0092_PP). D.P. was supported by a fellowship from Junta de Castilla y León co-financed by the European Social Fund. E.N. was supported by Fundación de Investigación Sanitaria of León. M.V.G.M. was supported by CIBERehd contracts. CIBERehd is funded by the Instituto de Salud Carlos III, Spai

Beneficial effects of exercise on gut microbiota functionality and barrier integrity, and gut-liver axis crosstalk in an "in vivo" model of early obesity and non-alcoholic fatty liver disease

[EN]Childhood obesity has reached epidemic levels, representing one of the most serious public health concerns associated with metabolic syndrome and non-alcoholic fatty liver disease (NAFLD). There is limited clinical experience concerning pediatric NAFLD patients, and thus the therapeutic options are scarce. The aim of this study was to evaluate the benefits of exercise on gut microbiota composition and functionality balance, and consequent effects on early obesity and NAFLD onset in an in vivo model. Juvenile (21-day-old) male Wistar rats fed a control diet or a high-fat diet (HFD) were subjected to a combined aerobic and resistance training protocol. Fecal microbiota was sequenced by an Illumina MiSeq system, and parameters related to metabolic syndrome, fecal metabolome, intestinal barrier integrity, bile acid metabolism and transport, and alteration of the gut-liver axis were measured. Exercise decreased HFD-induced body weight gain, metabolic syndrome and hepatic steatosis, as a result of its lipid metabolism modulatory capacity. Gut microbiota composition and functionality were substantially modified as a consequence of diet, age and exercise intervention. In addition, the training protocol increased Parabacteroides, Bacteroides and Flavobacterium genera, correlating with a beneficial metabolomic profile, whereas Blautia, Dysgonomonas and Porphyromonas showed an opposite pattern. Exercise effectively counteracted HFD-induced microbial imbalance, leading to intestinal barrier preservation, which, in turn, prevented deregulation of the gut-liver axis and improved bile acid homeostasis, determining the clinical outcomes of NAFLD. In conclusion, we provide scientific evidence highlighting the benefits of gut microbiota composition and functionality modulation by physical exercise protocols in the management of early obesity and NAFLD development.SIThis work was supported by grants from Ministerio de Economıa y Competitividad ́ (BFU2017-87960-R), Junta de Castilla y León and the European Regional Development Fund (FEDER) (LE063U16 and GRS1888/A/18). D.P. and S.C.-P. were supported by a fellowship from Junta de Castilla y León, co-financed by the European Social Fund. E.N. was supported by Fundación de Investigación Sanitaria of León. M.V.G.-M. was supported by contracts from the CIBERehd, which is funded by Instituto de Salud Carlos III

Exercise training modulates the gut microbiota profile and impairs inflammatory signaling pathways in obese children

[EN] Childhood obesity has reached epidemic levels and is a serious health concern associated with metabolic syndrome, nonalcoholic fatty liver disease, and gut microbiota alterations. Physical exercise is known to counteract obesity progression and modulate the gut microbiota composition. This study aims to determine the effect of a 12-week strength and endurance combined training program on gut microbiota and inflammation in obese pediatric patients. Thirty-nine obese children were assigned randomly to the control or training group. Anthropometric and biochemical parameters, muscular strength, and inflammatory signaling pathways in mononuclear cells were evaluated. Bacterial composition and functionality were determined by massive sequencing and metabolomic analysis. Exercise reduced plasma glucose levels and increased dynamic strength in the upper and lower extremities compared with the obese control group. Metagenomic analysis revealed a bacterial composition associated with obesity, showing changes at the phylum, class, and genus levels. Exercise counteracted this profile, significantly reducing the Proteobacteria phylum and Gammaproteobacteria class. Moreover, physical activity tended to increase some genera, such as Blautia, Dialister, and Roseburia, leading to a microbiota profile similar to that of healthy children. Metabolomic analysis revealed changes in short-chain fatty acids, branched-chain amino acids, and several sugars in response to exercise, in correlation with a specific microbiota profile. Finally, the training protocol significantly inhibited the activation of the obesity-associated NLRP3 signaling pathway. Our data suggest the existence of an obesity-related deleterious microbiota profile that is positively modified by physical activity intervention. Exercise training could be considered an efficient nonpharmacological therapy, reducing inflammatory signaling pathways induced by obesity in children via microbiota modulation.This work was supported by grants from Ministerio de Economía, Industria y Competitividad (BFU2017–87960-R), Junta de Castilla y León and the European Regional Development Fund (FEDER) (LE063U16 and GRS1888/A/18). CIBERehd is funded by the Instituto de Salud Carlos III, Spain. B.E and M.J.F were supported by a fellowship from Ministerio de Educación (FPU15/05051 and FPU18/06257). E.N. was supported by Fundación de Investigación Sanitaria of León. D.P. was supported by a fellowship from Junta de Castilla y León, cofinanced by the European Social Fund

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

Modulación de la microbiota intestinal en el tratamiento de la enfermedad de hígado graso no alcohólico y síndrome metabólico. Estudio de un tratamiento experimental con quercetina y de la transferencia de microbiota a ratones libres de gérmenes = Intestinal microbiota modulation in the treatment of non-alcoholic fatty liver disease and metabolic syndrome. Study of an experimental treatment with quercetin and intestinal microbiota transfer to germ-free mice

215 p.La enfermedad de hígado graso no alcohólico (NAFLD) es actualmente la enfermedad hepática crónica más extendida y presenta, además, una tendencia al alza que refleja su estrecha asociación con otras patologías ampliamente distribuidas como la obesidad, la diabetes mellitus tipo II y el síndrome metabólico. Si bien la denominación de NAFLD hace referencia a una enfermedad, el término se utiliza para dar cabida a un espectro de patologías hepáticas que comienzan con la esteatosis o acumulación de grasa en el hígado y avanza hacia estadios progresivamente más graves como la esteatohepatitis no alcohólica, pudiendo desembocar en la aparición de cirrosis e incluso carcinoma hepatocelular. De esta manera la patología se asocia a una importante morbilidad y mortalidad, en muchos casos ligada a la aparición de episodios cardiometabólicos o al desarrollo de procesos de malignidad, siendo además cada vez más relevante como causa de trasplante hepático, lo que supone en definitiva un notable impacto sobre los sistemas sanitarios. Una de las principales dificultades a la hora de abordar la enfermedad de NAFLD es la complejidad de los factores implicados en su desarrollo. Según la hipótesis más aceptada actualmente, el modelo del múltiple impacto, son varios los procesos que actuando de forma simultánea y concurrente desencadenan la enfermedad, considerando entre ellos la resistencia a la insulina, la disfunción de orgánulos celulares como fuentes de estrés o la aparición de procesos inflamatorios. Junto a ellos se ha destacado recientemente el papel que podría desempeñar la microbiota intestinal en las enfermedades hepáticas y metabólicas, interactuando a través del eje intestino-hígado con los factores mencionados anteriormente. La microbiota intestinal agrupa a los numerosos microorganismos, fundamentalmente bacterias, que pueblan el tracto gastrointestinal y realiza funciones fundamentales para el hospedador. De esta manera alteraciones en su composición, condición conocida como disbiosis, podría contribuir de forma destacada a la patogénesis de distintas enfermedades. Por otra parte, la ausencia de un tratamiento de probada eficacia para NAFLD atrae la atención sobre nuevas estrategias terapéuticas. En este sentido la quercetina, podría ser una prometedora alternativa no sólo por las conocidas propiedades antioxidantes y antiinflamatorias del grupo de los flavonoides sino también por una posible acción prebiótica sobre la microbiota intestinal. Asimismo, la posibilidad de revertir la situación de disbiosis mediante el trasplante de microbiota intestinal de un donante sano podría suponer una estrategia efectiva. Por todo ello planteamos profundizar en los mecanismos patogénicos implicados en el desarrollo de la enfermedad de hígado graso no alcohólico asociada a obesidad, con un enfoque basado en el papel de la microbiota intestinal como factor clave en la aparición de la patología y como una potencial diana terapéutica. Para ello abordamos en primer lugar, un estudio sobre la utilidad del flavonol quercetina para frenar la progresión de la enfermedad en un modelo in vivo de obesidad, síndrome metabólico y NAFLD, entre otros mecanismos, por su posible acción prebiótica sobre la composición de la microbiota intestinal. En una etapa posterior, sobre un modelo realizado en ratones libres de gérmenes tratamos de evaluar en qué medida la transferencia de microbiota de donantes seleccionados puede determinar la susceptibilidad al desarrollo de la patología, señalando los posibles mecanismos responsables de dicha respuesta En el modelo in vivo basado en la dieta rica en grasa la suplementación con quercetina fue capaz de revertir o atenuar los principales mecanismos patogénicos de la enfermedad, incluyendo la resistencia a la insulina, el estrés oxidativo y de retículo endoplásmico, la alteración del metabolismo lipídico hepático o el desarrollo de inflamación. Asimismo, la disbiosis intestinal, la alteración de la permeabilidad intestinal y del eje intestino hígado inducidos por la dieta fueron compensados por el efecto de la quercetina. En conjunto la modulación de estos mecanismos logró disminuir el depósito intrahepático de lípidos y las demás alteraciones metabólicas asociadas a NAFLD y síndrome metabólico. La colonización de ratones libres de gérmenes con la microbiota de donantes seleccionados del modelo anterior logró transferir el fenotipo metabólico asociado, de manera que la microbiota de un donante no respondedor a la dieta rica en grasa o del donante suplementado con quercetina ejerció un efecto protector frente al desarrollo de la enfermedad de hígado graso, relacionado con diferencias en cuanto a la abundancia de géneros como Helicobacter o Akkermansia. Por el contrario, la colonización con microbiota de un donante respondedor manifestó un efecto predisponente asociado a una alteración del eje intestino-hígado y la activación de receptores tipo toll y tipo nod a nivel hepático. El análisis en profundidad de los perfiles de microbiota protector y no protector frente a NAFLD señaló importantes diferencias relacionadas con la síntesis, abundancia y circulación enterohepática de los ácidos biliares y con la síntesis y captación de lípidos. Así, el perfil protector manifestó una mayor capacidad para producir ácidos biliares secundarios, cuya circulación enterohepática se vio además estimulada en este grupo. Por el contrario, el perfil no protector manifestó una menor conversión de ácidos biliares primarios junto con una promoción de procesos relacionados con el depósito lipídico hepático como la captación de ácidos grasos libres y la lipogéneis de novo. Por todo ello cabe concluir que la microbiota intestinal ejerce un papel fundamental en el desarrollo de síndrome metabólico y enfermedad de hígado graso no alcohólico. La administración del flavonol quercetina o el trasplante de microbiota de donantes con características determinadas manifiestan efectos beneficiosos frente a estas patologías a través de la modulación de la microbiota y deben ser, por tanto, consideradas como alternativa terapéutica en el manejo de la enfermeda

Novel genes and sex differences in COVID-19 severity.

Here we describe the results of a genome-wide study conducted in 11 939 COVID-19 positive cases with an extensive clinical information that were recruited from 34 hospitals across Spain (SCOURGE consortium). In sex-disaggregated genome-wide association studies for COVID-19 hospitalization, genome-wide significance (p < 5x10-8) was crossed for variants in 3p21.31 and 21q22.11 loci only among males (p = 1.3x10-22 and p = 8.1x10-12, respectively), and for variants in 9q21.32 near TLE1 only among females (p = 4.4x10-8). In a second phase, results were combined with an independent Spanish cohort (1598 COVID-19 cases and 1068 population controls), revealing in the overall analysis two novel risk loci in 9p13.3 and 19q13.12, with fine-mapping prioritized variants functionally associated with AQP3 (p = 2.7x10-8) and ARHGAP33 (p = 1.3x10-8), respectively. The meta-analysis of both phases with four European studies stratified by sex from the Host Genetics Initiative confirmed the association of the 3p21.31 and 21q22.11 loci predominantly in males and replicated a recently reported variant in 11p13 (ELF5, p = 4.1x10-8). Six of the COVID-19 HGI discovered loci were replicated and an HGI-based genetic risk score predicted the severity strata in SCOURGE. We also found more SNP-heritability and larger heritability differences by age (<60 or ≥ 60 years) among males than among females. Parallel genome-wide screening of inbreeding depression in SCOURGE also showed an effect of homozygosity in COVID-19 hospitalization and severity and this effect was stronger among older males. In summary, new candidate genes for COVID-19 severity and evidence supporting genetic disparities among sexes are provided

GWAS and meta-analysis identifies 49 genetic variants underlying critical COVID-19

Data availability: Downloadable summary data are available through the GenOMICC data site (https://genomicc.org/data). Summary statistics are available, but without the 23andMe summary statistics, except for the 10,000 most significant hits, for which full summary statistics are available. The full GWAS summary statistics for the 23andMe discovery dataset will be made available through 23andMe to qualified researchers under an agreement with 23andMe that protects the privacy of the 23andMe participants. For further information and to apply for access to the data, see the 23andMe website (https://research.23andMe.com/dataset-access/). All individual-level genotype and whole-genome sequencing data (for both academic and commercial uses) can be accessed through the UKRI/HDR UK Outbreak Data Analysis Platform (https://odap.ac.uk). A restricted dataset for a subset of GenOMICC participants is also available through the Genomics England data service. Monocyte RNA-seq data are available under the title ‘Monocyte gene expression data’ within the Oxford University Research Archives (https://doi.org/10.5287/ora-ko7q2nq66). Sequencing data will be made freely available to organizations and researchers to conduct research in accordance with the UK Policy Framework for Health and Social Care Research through a data access agreement. Sequencing data have been deposited at the European Genome–Phenome Archive (EGA), which is hosted by the EBI and the CRG, under accession number EGAS00001007111.Extended data figures and tables are available online at https://www.nature.com/articles/s41586-023-06034-3#Sec21 .Supplementary information is available online at https://www.nature.com/articles/s41586-023-06034-3#Sec22 .Code availability: Code to calculate the imputation of P values on the basis of SNPs in linkage disequilibrium is available at GitHub (https://github.com/baillielab/GenOMICC_GWAS).Acknowledgements: We thank the members of the Banco Nacional de ADN and the GRA@CE cohort group; and the research participants and employees of 23andMe for making this work possible. A full list of contributors who have provided data that were collated in the HGI project, including previous iterations, is available online (https://www.covid19hg.org/acknowledgements).Change history: 11 July 2023: A Correction to this paper has been published at: https://doi.org/10.1038/s41586-023-06383-z. -- In the version of this article initially published, the name of Ana Margarita Baldión-Elorza, of the SCOURGE Consortium, appeared incorrectly (as Ana María Baldion) and has now been amended in the HTML and PDF versions of the article.Copyright © The Author(s) 2023, Critical illness in COVID-19 is an extreme and clinically homogeneous disease phenotype that we have previously shown1 to be highly efficient for discovery of genetic associations2. Despite the advanced stage of illness at presentation, we have shown that host genetics in patients who are critically ill with COVID-19 can identify immunomodulatory therapies with strong beneficial effects in this group3. Here we analyse 24,202 cases of COVID-19 with critical illness comprising a combination of microarray genotype and whole-genome sequencing data from cases of critical illness in the international GenOMICC (11,440 cases) study, combined with other studies recruiting hospitalized patients with a strong focus on severe and critical disease: ISARIC4C (676 cases) and the SCOURGE consortium (5,934 cases). To put these results in the context of existing work, we conduct a meta-analysis of the new GenOMICC genome-wide association study (GWAS) results with previously published data. We find 49 genome-wide significant associations, of which 16 have not been reported previously. To investigate the therapeutic implications of these findings, we infer the structural consequences of protein-coding variants, and combine our GWAS results with gene expression data using a monocyte transcriptome-wide association study (TWAS) model, as well as gene and protein expression using Mendelian randomization. We identify potentially druggable targets in multiple systems, including inflammatory signalling (JAK1), monocyte–macrophage activation and endothelial permeability (PDE4A), immunometabolism (SLC2A5 and AK5), and host factors required for viral entry and replication (TMPRSS2 and RAB2A).GenOMICC was funded by Sepsis Research (the Fiona Elizabeth Agnew Trust), the Intensive Care Society, a Wellcome Trust Senior Research Fellowship (to J.K.B., 223164/Z/21/Z), the Department of Health and Social Care (DHSC), Illumina, LifeArc, the Medical Research Council, UKRI, a BBSRC Institute Program Support Grant to the Roslin Institute (BBS/E/D/20002172, BBS/E/D/10002070 and BBS/E/D/30002275) and UKRI grants MC_PC_20004, MC_PC_19025, MC_PC_1905 and MRNO2995X/1. A.D.B. acknowledges funding from the Wellcome PhD training fellowship for clinicians (204979/Z/16/Z), the Edinburgh Clinical Academic Track (ECAT) programme. This research is supported in part by the Data and Connectivity National Core Study, led by Health Data Research UK in partnership with the Office for National Statistics and funded by UK Research and Innovation (grant MC_PC_20029). Laboratory work was funded by a Wellcome Intermediate Clinical Fellowship to B.F. (201488/Z/16/Z). We acknowledge the staff at NHS Digital, Public Health England and the Intensive Care National Audit and Research Centre who provided clinical data on the participants; and the National Institute for Healthcare Research Clinical Research Network (NIHR CRN) and the Chief Scientist’s Office (Scotland), who facilitate recruitment into research studies in NHS hospitals, and to the global ISARIC and InFACT consortia. GenOMICC genotype controls were obtained using UK Biobank Resource under project 788 funded by Roslin Institute Strategic Programme Grants from the BBSRC (BBS/E/D/10002070 and BBS/E/D/30002275) and Health Data Research UK (HDR-9004 and HDR-9003). UK Biobank data were used in the GSMR analyses presented here under project 66982. The UK Biobank was established by the Wellcome Trust medical charity, Medical Research Council, Department of Health, Scottish Government and the Northwest Regional Development Agency. It has also had funding from the Welsh Assembly Government, British Heart Foundation and Diabetes UK. The work of L.K. was supported by an RCUK Innovation Fellowship from the National Productivity Investment Fund (MR/R026408/1). J.Y. is supported by the Westlake Education Foundation. SCOURGE is funded by the Instituto de Salud Carlos III (COV20_00622 to A.C., PI20/00876 to C.F.), European Union (ERDF) ‘A way of making Europe’, Fundación Amancio Ortega, Banco de Santander (to A.C.), Cabildo Insular de Tenerife (CGIEU0000219140 ‘Apuestas científicas del ITER para colaborar en la lucha contra la COVID-19’ to C.F.) and Fundación Canaria Instituto de Investigación Sanitaria de Canarias (PIFIISC20/57 to C.F.). We also acknowledge the contribution of the Centro National de Genotipado (CEGEN) and Centro de Supercomputación de Galicia (CESGA) for funding this project by providing supercomputing infrastructures. A.D.L. is a recipient of fellowships from the National Council for Scientific and Technological Development (CNPq)-Brazil (309173/2019-1 and 201527/2020-0)