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      Asociación entre metilación del ADN y tabaquismo en pacientes con isquemia crónica amenazante de extremidades inferiores Translated title: Association between DNA methylation and smoking in patients with chronic limb-threatening ischemia

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          Abstract

          Resumen Introducción y objetivo: estudiar la asociación entre tabaquismo y el nivel de metilación de dos regiones genómicas en pacientes con enfermedad arterial periférica (EAP). Método: estudio transversal de 297 pacientes (edad media: 69,6 años; varones: 78,5 %) diagnosticados de isquemia crónica de extremidades inferiores en diferentes estadios clínicos entre marzo de 2016 y diciembre de 2019 en el servicio de cirugía vascular del Hospital del Mar (Barcelona). Se analizó la metilación de Cg02156642 y de Cg03636183 asociados en otros estudios al tabaquismo. Se realizó una regresión lineal múltiple para discriminar las variables asociadas al nivel de metilación. Se calculó el área bajo la curva ROC para discriminar el nivel de metilación entre fumadores y no fumadores. Resultados: de la muestra, 46 pacientes (15,5 %) eran no fumadores; 132 (44,4 %), exfumadores y 119 (40,1 %), fumadores. No se observó una asociación entre la exposición al tabaco y el nivel de metilación del Cg02156642, pero sí con el de Cg03636183: los fumadores presentaban menor nivel de metilación y, además, a más carga de tabaco menos metilación (Rho de Spearman: -0,324; p < 0,001). Un nivel de metilación en este CpG del 80 % tiene una sensibilidad (S) del 90,0 % y una especificidad (E) del 83,5 % para discriminar entre fumadores y nunca fumadores. Para discriminar entre fumadores y exfumadores, un nivel de metilación del 75 % tiene una S del 69 % y una E del 56,9 %. Al ajustar por todas las variables relacionadas con la metilación, la magnitud de esta asociación entre Cg03636183 y tabaquismo se mantenía significativa entre los nunca fumadores y los fumadores. Conclusiones: la metilación del cpg cg03636183 se asocia a tabaquismo en pacientes con eap y está directamente relacionada con la carga de tabaco. Este biomarcador podría utilizarse en la práctica clínica para valorar el consumo de tabaco de nuestros pacientes.

          Translated abstract

          Abstract Introduction and objective: to study the association between smoking and the methylation level of 2 genomic regions in patients with peripheral artery disease (PAD). Method: cross-sectional study of 297 patients (mean age, 69.6 years; males, 78.5%) diagnosed with chronic lower extremity ischemia at various clinical stages from March 2016 through December 2019 at the Vascular Surgery Unit of Hospital del Mar, Barcelona, Catalonia, Spain. Methylation analysis of Cg02156642 and Cg03636183, previously associated with smoking in former studies was performed. Multiple linear regression was conducted to identify variables associated with methylation levels. The area under the ROC curve was estimated to discriminate methylation levels between smokers and non-smokers. Results: among the sample, 46 patients (15.5%) were non-smokers, 132 (44.4%) were former smokers, and 119 (40.1%) were current smokers. No association was seen between tobacco exposure and methylation levels of Cg02156642. However, an association was found with Cg03636183: smokers had lower methylation levels, and a higher smoking load was associated with lower methylation (Spearman's Rho, -0.324; p < .001). A methylation level of 80% in this region showed a 90.0% sensitivity and an 83.5% specificity to discriminate between smokers and never smokers. To discriminate between smokers and former smokers, a methylation level of 75% had an 69% sensitivity and an 56.9% specificity. After adjusting for all variables associated with methylation, the association between Cg03636183 and smoking remained significant among never smokers and smokers. Conclusions: methylation of the Cg03636183 region is associated with smoking in patients with PAD and is directly associated with the smoking load. This biomarker could be used in the routine clinical practice to assess tobacco use in our patients.

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          Most cited references28

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          Prevalence of and risk factors for peripheral arterial disease in the United States: results from the National Health and Nutrition Examination Survey, 1999-2000.

          Peripheral arterial disease (PAD) is associated with significant morbidity and mortality and is an important marker of subclinical coronary heart disease. However, estimates of PAD prevalence in the general US population have varied widely. We analyzed data from 2174 participants aged 40 years and older from the 1999-2000 National Health and Nutrition Examination Survey. PAD was defined as an ankle-brachial index <0.90 in either leg. The prevalence of PAD among adults aged 40 years and over in the United States was 4.3% (95% CI 3.1% to 5.5%), which corresponds to approximately 5 million individuals (95% CI 4 to 7 million). Among those aged 70 years or over, the prevalence was 14.5% (95% CI 10.8% to 18.2%). In age- and gender-adjusted logistic regression analyses, black race/ethnicity (OR 2.83, 95% CI 1.48 to 5.42) current smoking (OR 4.46, 95% CI 2.25 to 8.84), diabetes (OR 2.71, 95% CI 1.03 to 7.12), hypertension (OR 1.75, 95% CI 0.97 to 3.13), hypercholesterolemia (OR 1.68, 95% CI 1.09 to 2.57), and low kidney function (OR 2.00, 95% CI 1.08 to 3.70) were positively associated with prevalent PAD. More than 95% of persons with PAD had 1 or more cardiovascular disease risk factors. Elevated fibrinogen and C-reactive protein levels were also associated with PAD. This study provides nationally representative prevalence estimates of PAD in the United States, revealing that PAD affects more than 5 million adults. PAD prevalence increases dramatically with age and disproportionately affects blacks. The vast majority of individuals with PAD have 1 or more cardiovascular disease risk factors that should be targeted for therapy.
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            Differences in smoking associated DNA methylation patterns in South Asians and Europeans

            Background DNA methylation is strongly associated with smoking status at multiple sites across the genome. Studies have largely been restricted to European origin individuals yet the greatest increase in smoking is occurring in low income countries, such as the Indian subcontinent. We determined whether there are differences between South Asians and Europeans in smoking related loci, and if a smoking score, combining all smoking related DNA methylation scores, could differentiate smokers from non-smokers. Results Illumina HM450k BeadChip arrays were performed on 192 samples from the Southall And Brent REvisited (SABRE) cohort. Differential methylation in smokers was identified in 29 individual CpG sites at 18 unique loci. Interaction between smoking status and ethnic group was identified at the AHRR locus. Ethnic differences in DNA methylation were identified in non-smokers at two further loci, 6p21.33 and GNG12. With the exception of GFI1 and MYO1G these differences were largely unaffected by adjustment for cell composition. A smoking score based on methylation profile was constructed. Current smokers were identified with 100% sensitivity and 97% specificity in Europeans and with 80% sensitivity and 95% specificity in South Asians. Conclusions Differences in ethnic groups were identified in both single CpG sites and combined smoking score. The smoking score is a valuable tool for identification of true current smoking behaviour. Explanations for ethnic differences in DNA methylation in association with smoking may provide valuable clues to disease pathways.
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              Human aging-associated DNA hypermethylation occurs preferentially at bivalent chromatin domains.

              There is a growing realization that some aging-associated phenotypes/diseases have an epigenetic basis. Here, we report the first genome-scale study of epigenomic dynamics during normal human aging. We identify aging-associated differentially methylated regions (aDMRs) in whole blood in a discovery cohort, and then replicate these aDMRs in sorted CD4(+) T-cells and CD14(+) monocytes in an independent cohort, suggesting that aDMRs occur in precursor haematopoietic cells. Further replication of the aDMRs in buccal cells, representing a tissue that originates from a different germ layer compared with blood, demonstrates that the aDMR signature is a multitissue phenomenon. Moreover, we demonstrate that aging-associated DNA hypermethylation occurs predominantly at bivalent chromatin domain promoters. This same category of promoters, associated with key developmental genes, is frequently hypermethylated in cancers and in vitro cell culture, pointing to a novel mechanistic link between aberrant hypermethylation in cancer, aging, and cell culture.
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                Author and article information

                Journal
                angiologia
                Angiología
                Angiología
                Arán Ediciones S.L. (Madrid, Madrid, Spain )
                0003-3170
                1695-2987
                June 2023
                : 75
                : 3
                : 146-154
                Affiliations
                [4] Málaga orgnameParque Tecnológico de Andalucía (PTA) orgdiv1ECAI de Genómica España
                [3] Barcelona Cataluña orgnameUniversitat Pompeu Fabra orgdiv1Departament de Medicina i Ciències de la Vida Spain
                [5] Barcelona orgnameUniversidad Central de Cataluña (Uvic-UCC) orgdiv1Universidad de VIC orgdiv2Departamento de Medicina España
                [2] Barcelona orgnameInstitut Hospital del Mar d'Investigacions Mèdiques (IMIM) España
                [1] Barcelona orgnameHospital del Mar orgdiv1Cirugía Vascular y Endovascular orgdiv2Servicio de Angiología España
                Article
                S0003-31702023000300003 S0003-3170(23)07500300003
                10.20960/angiologia.00499
                005c90d0-2513-4f91-876d-0c06715f5cf3

                This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.

                History
                : 23 January 2023
                : 09 February 2023
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 28, Pages: 9
                Product

                SciELO Spain

                Categories
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                Tabaquismo,Isquemia crónica amenazante de miembros inferiores,Metilación,Smoking,Chronic limb-threatening ischemia,Methylation

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