Identificación de factores genéticos asociados con el desarrollo de la microalbuminuria en la hipertensión esencial

IDENTIFICACIÓN DE FACTORES GENÉTICOS ASOCIADOS CON EL DESARROLLO DE LA MICROALBUMINURIA EN LA HIPERTENSIÓN ESENCIAL La excreción urinaria de albúmina es uno de los marcadores de riesgo más utilizados en el estudio del desarrollo de la enfermedad cardiovascular. El incremento de los valores de EUA está provocado por el aumento de la filtración glomerular a nivel del túbulo proximal, la cual puede estar producida por un aumento sistemático de la presión arterial. Tradicionalmente se ha venido estudiando la presencia de microalbuminuria en relación a otros factores de riesgo cardiovascular como son la diabetes mellitus y la hipertensión, debido la relevacia de estas patologías en el desarrollo de daño orgánico. Existen múltiples estudios que buscan establecer una base genética en el incremento de los valores de excreción urinaria de albúmina asociado a la hipertensión esencial. Hipotesis y Objetivos: Existe una base genética de rasgos complejos en el incremento de los niveles de excreción urinaria de albúmina asociados en la hipertensión. Nuestro grupo ha realizado un GWAS en población hipertensa con el objetivo de identificar una serie de marcadores involucrados en el incremento de la excreción urinaria de albúmina. Nuestro objetivo actual es verificar las asociaciones encontradas, así como otras asociaciones descritas para otras poblaciones. Materiales y Métodos: Se llevó a cabo el genotipado de 1.536 SNPS, procedentes del GWAS realizado previamente por nuestro grupo, en una nueva población hipertensa de 910 individuos utilizando la tecnología Infinium GoldenGate Genotyping Assay de Illumina, a partir de los resultados obtenidos, se validaron mediante secuenciación NGS en la población del Estudio [email protected] (n = 4.785) los 20 SNPs más significativos en su asociación con las variables relacionadas con la excreción urinaria de albúmina. Para el estudio de secuenciación se utilizó la tecnología paired-end 150x2 de Illumina. Resultados: Los análisis realizados dieron como resultado la asociación de varios loci con las variables relacionadas con la excreción urinaria de albúmina en la población general: rs6972824 (p.valor 0.015) y rs2820950 (p.valor 0.016). También se obtuvieron resultados para la asociación con la enfermedad renal crónica de los polimorfismos rs13232567 (p.valor 1.03e-3) y rs12322500 (p.valor 0.037). Así mismo se realizaron los análisis en la población hipertensa en la cual se asoció con los valores de excreción urinaria de albúmina, cuantitativa o cualitativamente los polimorfismos rs3114316 (p.valor 7.35e-3), rs3114316 (p.valor 0.031), rs6972824 (p.valor 0.014) y rs9950398 (p.valor 0.030), asociándose también con los valores de presión arterial. Conclusiones: Según nuestros estudios, diferentes marcadores genéticos se asocian con los valores de excreción urinaria de albúmina, además algunos de ellos se han asociado a su vez con los valores de presión arterial. Estos resultados ponen de manifiesto la relación genética entre los valores de excreción urinaria de albúmina y presión arterial.IDENTIFICATION OF GENETIC FACTORS ASSOCIATED WITH THE DEVELOPMENT OF MICROALBUMINURIA IN ESSENTIAL HYPERTENSION Urinary albumin excretion is one of the most commonly used risk markers in the development of cardiovascular disease. High urinary albumin excretion values are caused by an increase of the glomerular filtration at the proximal tubule, which may be caused by a systematic increase in blood pressure. The presence of microalbuminuria has traditionally been studied in relation to other cardiovascular risk factors such as diabetes mellitus and hypertension, due to the relevance of these pathologies in the development of organic damage. There are multiple studies that seek to establish a genetic basis in the increase of urinary albumin excretion values associated with hypertension. Hypothesis and Objectives: There is a genetic basis of complex traits in increasing urinary albumin excretion levels associated with hypertension. Our group has performed a GWAS in a hypertensive population with the goal of identifying different markers involved in the increase of the urinary albumin excretion. Our current objective is to verify the associations found previously, as well as other associations described for other populations. Materials and Methods: Genotyping of 1,536 SNPS from the GWAS previously performed by our group was carried out in a hypertensive population of 910 individuals using the Infinium GoldenGate Genotyping Assay technology from Illumina. From the results obtained, we validated by NGS sequencing in the population of the [email protected] Study (n = 4,785) the 20 most significant SNPs in their association with urinary albumin excretion-related variables. For the sequencing study, the technology was NGS paired-end 150x2 from Illumina. Results: The analysis performed resulted in the association of several loci with urinary albumin excretion -related variables in the general population: rs6972824 (p.value 0.015) and rs2820950 (p.value 0.016). Results were also obtained for the association between chronic kidney disease and rs13232567 (p.value 1.03e-3) and rs12322500 (p.value 0.037) polymorphisms. We also performed analysis in the hypertensive population of [email protected] Study in which we observed association between the urinary albumin excretion levels, as a quantitative or qualitative trait, and the polymorphisms rs3114316 (p.value 7.35e-3), rs3114316 (p.value 0.031), rs6972824 (p.value) 0.014) and rs9950398 (p.value 0.030). These SNPs were also associated with blood pressure values. Conclusions: According to our studies, different genetic markers are associated with urinary albumin excretion values, in addition, some of them have been associated with blood pressure values. These results highlight the genetic basis of the relationship between urinary albumin excretion and blood pressure levels

Cadmium, Smoking, and Human Blood DNA Methylation Profiles in Adults from the Strong Heart Study.

The epigenetic effects of individual environmental toxicants in tobacco remain largely unexplored. Cadmium (Cd) has been associated with smoking-related health effects, and its concentration in tobacco smoke is higher in comparison with other metals. We studied the association of Cd and smoking exposures with human blood DNA methylation (DNAm) profiles. We also evaluated the implication of findings to relevant methylation pathways and the potential contribution of Cd exposure from smoking to explain the association between smoking and site-specific DNAm. We conducted an epigenome-wide association study of urine Cd and self-reported smoking (current and former vs. never, and cumulative smoking dose) with blood DNAm in 790,026 CpGs (methylation sites) measured with the Illumina Infinium Human MethylationEPIC (Illumina Inc.) platform in 2,325 adults 45-74 years of age who participated in the Strong Heart Study in 1989-1991. In a mediation analysis, we estimated the amount of change in DNAm associated with smoking that can be independently attributed to increases in urine Cd concentrations from smoking. We also conducted enrichment analyses and in silico protein-protein interaction networks to explore the biological relevance of the findings. At a false discovery rate (FDR)-corrected level of 0.05, we found 6 differentially methylated positions (DMPs) for Cd; 288 and 17, respectively, for current and former smoking status; and 77 for cigarette pack-years. Enrichment analyses of these DMPs displayed enrichment of 58 and 6 Gene Ontology and Kyoto Encyclopedia of Genes and Genomes gene sets, respectively, including biological pathways for cancer and cardiovascular disease. In in silico protein-to-protein networks, we observed key proteins in DNAm pathways directly and indirectly connected to Cd- and smoking-DMPs. Among DMPs that were significant for both Cd and current smoking (annotated to PRSS23, AHRR, F2RL3, RARA, and 2q37.1), we found statistically significant contributions of Cd to smoking-related DNAm. Beyond replicating well-known smoking epigenetic signatures, we found novel DMPs related to smoking. Moreover, increases in smoking-related Cd exposure were associated with differential DNAm. Our integrative analysis supports a biological link for Cd and smoking-associated health effects, including the possibility that Cd is partly responsible for smoking toxicity through epigenetic changes. https://doi.org/10.1289/EHP6345.This work was supported by grants by the National Heart, Lung, and Blood Institute (NHLBI) (under contract numbers 75N92019D00027, 75N92019D00028, 75N92019D00029, & 75N92019D00030) and previous grants (R01HL090863, R01HL109315, R01HL109301, R01HL109284, R01HL109282, and R01HL109319 and cooperative agreements U01HL41642, U01HL41652, U01HL41654, U01HL65520, and U01HL65521), by the National Institute of Health Sciences (R01ES021367, R01ES025216, P42ES010349, P30ES009089), by the Spanish Funds for Research In Health Sciences, Carlos III Health Institute, co-funded by European Regional Development Fund (CP12/03080 and PI15/00071), by Chilean CONICYT/FONDECYT-POSTDOCTORADO Nº3180486 (A.L.R.-C) and a fellowship from “La Caixa” Foundation (ID 100010434). The fellowship code is “LCF/BQ/DR19/11740016.”S

Arsenic exposure and human blood DNA methylation and hydroxymethylation profiles in two diverse populations from Bangladesh and Spain

This work was supported by the Strategic Action for Research in Health sciences (PI15/00071) and CIBERCV, which are initiatives from Instituto de Salud Carlos III and the Spanish Ministry of Science and Innovation and co-funded with European Funds for Regional Development (FEDER) , and the United States National Institute of Health (grants R01 CA133595, P30 ES009089, T32 ES007322, P42 ES010349, and F31 ES029019) . MTP was supported by the Third AstraZeneca Award for Spanish Young Researchers. ADR received the support of a fellowship from "la Caixa" Foundation (ID 100010434, fellowship code "LCF/BQ/DR19/11740016") .Domingo-Relloso, A.; Bozack, A.; Kiihl, S.; Rodriguez-Hernandez, Z.; Rentero-Garrido, P.; Casasnovas, JA.; Leon-Latre, M.... (2022). Arsenic exposure and human blood DNA methylation and hydroxymethylation profiles in two diverse populations from Bangladesh and Spain. Environmental Research. 204:1-12. https://doi.org/10.1016/j.envres.2021.11202111220

"Proteotranscriptomic analysis of advanced colorectal cancer patient derived organoids for drug sensitivity prediction"

Background: Patient-derived organoids (PDOs) from advanced colorectal cancer (CRC) patients could be a key platform to predict drug response and discover new biomarkers. We aimed to integrate PDO drug response with multi-omics characterization beyond genomics. Methods: We generated 29 PDO lines from 22 advanced CRC patients and provided a morphologic, genomic, and transcriptomic characterization. We performed drug sensitivity assays with a panel of both standard and non-standard agents in five long-term cultures, and integrated drug response with a baseline proteomic and transcriptomic characterization by SWATH-MS and RNA-seq analysis, respectively. Results: PDOs were successfully generated from heavily pre-treated patients, including a paired model of advanced MSI high CRC deriving from pre- and post-chemotherapy liver metastasis. Our PDOs faithfully reproduced genomic and phenotypic features of original tissue. Drug panel testing identified differential response among PDOs, particularly to oxaliplatin and palbociclib. Proteotranscriptomic analyses revealed that oxaliplatin non-responder PDOs present enrichment of the t-RNA aminoacylation process and showed a shift towards oxidative phosphorylation pathway dependence, while an exceptional response to palbociclib was detected in a PDO with activation of MYC and enrichment of chaperonin T-complex protein Ring Complex (TRiC), involved in proteome integrity. Proteotranscriptomic data fusion confirmed these results within a highly integrated network of functional processes involved in differential response to drugs. Conclusions: Our strategy of integrating PDOs drug sensitivity with SWATH-mass spectrometry and RNA-seq allowed us to identify different baseline proteins and gene expression profiles with the potential to predict treatment response/resistance and to help in the development of effective and personalized cancer therapeutics

Electronic Supplementary Material 2: Summary of 407 genes associated to subclinical atherosclerosis-DMRs, including gene location, reported interactions with BED effector proteins and associated ANN scores from <i>In silico</i> epigenetics of metal exposure and subclinical atherosclerosis in middle aged men: pilot results from the Aragon Workers Health Study

We explored the association of metal levels with subclinical atherosclerosis and epigenetic changes in relevant biological pathways. Whole blood DNA Infinium Methylation 450 K data were obtained from 23 of 73 middle age men without clinically evident cardiovascular disease who participated in the Aragon Workers Health Study in 2009 (baseline visit) and had available baseline urinary metals and subclinical atherosclerosis measures obtained in 2010–2013 (follow-up visit). The median metal levels were 7.36 µg g⁻¹, 0.33 µg g⁻¹, 0.11 µg g⁻¹ and 0.07 µg g⁻¹, for arsenic (sum of inorganic and methylated species), cadmium, antimony and tungsten, respectively. Urine cadmium and tungsten were associated with femoral and carotid intima-media thickness, respectively (Pearson's <i>r</i> = 0.27; <i>p</i> = 0.03 in both cases). Among nearest genes to identified differentially methylated regions (DMRs), 46% of metal-DMR genes overlapped with atherosclerosis-DMR genes (<i>p</i> < 0.001). Pathway enrichment analysis of atherosclerosis-DMR genes showed a role in inflammatory, metabolic and transport pathways. In <i>in silico</i> protein-to-protein interaction networks among proteins encoded by 162 and 108 genes attributed to atherosclerosis- and metal-DMRs, respectively, with proteins known to have a role in atherosclerosis pathways, we observed hub proteins in the network associated with both atherosclerosis and metal-DMRs (e.g. <i>SMAD3</i> and <i>NOP56</i>), and also hub proteins associated with metal-DMRs only but with relevant connections with atherosclerosis effectors (e.g. <i>SSTR5</i>, <i>HDAC4</i>, <i>AP2A2</i>, <i>CXCL12</i> and <i>SSTR4</i>). Our integrative <i>in silico</i> analysis demonstrates the feasibility of identifying epigenomic regions linked to environmental exposures and potentially involved in relevant pathways for human diseases. While our results support the hypothesis that metal exposures can influence health due to epigenetic changes, larger studies are needed to confirm our pilot results.This article is part of a discussion meeting issue ‘Frontiers in epigenetic chemical biology’

Electronic Supplementary Material 3: Protein coding genes associated to subclinical atherosclerosis-DMRs included in the ANN analysis. from <i>In silico</i> epigenetics of metal exposure and subclinical atherosclerosis in middle age men: pilot results from the Aragon Workers Health Study

The ANN predictive score is expressed in percentage and reflects the relatedness between a given protein and reported atherosclerosis protein effectors or its motives (ANN score> 76% indicates a strong relatedness with an uncertainty p-value of <0.05; ANN score 40-75% indicates medium-strong relatedness with an uncertainty p-value of 0.05 - 0.25; ANN score <40% indicates weak relatedness with an uncertainty p-value > 0.25)

Electronic Supplementary Material 4: Summary of genes associated to metal-DMR, gene location, reported interactions with BED effector proteins and associated ANN scores. from <i>In silico</i> epigenetics of metal exposure and subclinical atherosclerosis in middle aged men: pilot results from the Aragon Workers Health Study

We explored the association of metal levels with subclinical atherosclerosis and epigenetic changes in relevant biological pathways. Whole blood DNA Infinium Methylation 450 K data were obtained from 23 of 73 middle age men without clinically evident cardiovascular disease who participated in the Aragon Workers Health Study in 2009 (baseline visit) and had available baseline urinary metals and subclinical atherosclerosis measures obtained in 2010–2013 (follow-up visit). The median metal levels were 7.36 µg g⁻¹, 0.33 µg g⁻¹, 0.11 µg g⁻¹ and 0.07 µg g⁻¹, for arsenic (sum of inorganic and methylated species), cadmium, antimony and tungsten, respectively. Urine cadmium and tungsten were associated with femoral and carotid intima-media thickness, respectively (Pearson's <i>r</i> = 0.27; <i>p</i> = 0.03 in both cases). Among nearest genes to identified differentially methylated regions (DMRs), 46% of metal-DMR genes overlapped with atherosclerosis-DMR genes (<i>p</i> < 0.001). Pathway enrichment analysis of atherosclerosis-DMR genes showed a role in inflammatory, metabolic and transport pathways. In <i>in silico</i> protein-to-protein interaction networks among proteins encoded by 162 and 108 genes attributed to atherosclerosis- and metal-DMRs, respectively, with proteins known to have a role in atherosclerosis pathways, we observed hub proteins in the network associated with both atherosclerosis and metal-DMRs (e.g. <i>SMAD3</i> and <i>NOP56</i>), and also hub proteins associated with metal-DMRs only but with relevant connections with atherosclerosis effectors (e.g. <i>SSTR5</i>, <i>HDAC4</i>, <i>AP2A2</i>, <i>CXCL12</i> and <i>SSTR4</i>). Our integrative <i>in silico</i> analysis demonstrates the feasibility of identifying epigenomic regions linked to environmental exposures and potentially involved in relevant pathways for human diseases. While our results support the hypothesis that metal exposures can influence health due to epigenetic changes, larger studies are needed to confirm our pilot results.This article is part of a discussion meeting issue ‘Frontiers in epigenetic chemical biology’

Electronic Supplementary Material 2: Summary of 407 genes associated to subclinical atherosclerosis-DMRs, including gene location, reported interactions with BED effector proteins and associated ANN scores from <i>In silico</i> epigenetics of metal exposure and subclinical atherosclerosis in middle age men: pilot results from the Aragon Workers Health Study

We explored the association of metal levels with subclinical atherosclerosis and epigenetic changes in relevant biological pathways. Whole blood DNA Infinium Methylation 450 K data were obtained from 23 of 73 middle age men without clinically evident cardiovascular disease who participated in the Aragon Workers Health Study in 2009 (baseline visit) and had available baseline urinary metals and subclinical atherosclerosis measures obtained in 2010–2013 (follow-up visit). The median metal levels were 7.36 µg g⁻¹, 0.33 µg g⁻¹, 0.11 µg g⁻¹ and 0.07 µg g⁻¹, for arsenic (sum of inorganic and methylated species), cadmium, antimony and tungsten, respectively. Urine cadmium and tungsten were associated with femoral and carotid intima-media thickness, respectively (Pearson's <i>r</i> = 0.27; <i>p</i> = 0.03 in both cases). Among nearest genes to identified differentially methylated regions (DMRs), 46% of metal-DMR genes overlapped with atherosclerosis-DMR genes (<i>p</i> < 0.001). Pathway enrichment analysis of atherosclerosis-DMR genes showed a role in inflammatory, metabolic and transport pathways. In <i>in silico</i> protein-to-protein interaction networks among proteins encoded by 162 and 108 genes attributed to atherosclerosis- and metal-DMRs, respectively, with proteins known to have a role in atherosclerosis pathways, we observed hub proteins in the network associated with both atherosclerosis and metal-DMRs (e.g. <i>SMAD3</i> and <i>NOP56</i>), and also hub proteins associated with metal-DMRs only but with relevant connections with atherosclerosis effectors (e.g. <i>SSTR5</i>, <i>HDAC4</i>, <i>AP2A2</i>, <i>CXCL12</i> and <i>SSTR4</i>). Our integrative <i>in silico</i> analysis demonstrates the feasibility of identifying epigenomic regions linked to environmental exposures and potentially involved in relevant pathways for human diseases. While our results support the hypothesis that metal exposures can influence health due to epigenetic changes, larger studies are needed to confirm our pilot results.This article is part of a discussion meeting issue ‘Frontiers in epigenetic chemical biology’

Electronic Supplementary Material 1: Supplementary Tables and Figures from <i>In silico</i> epigenetics of metal exposure and subclinical atherosclerosis in middle age men: pilot results from the Aragon Workers Health Study

We explored the association of metal levels with subclinical atherosclerosis and epigenetic changes in relevant biological pathways. Whole blood DNA Infinium Methylation 450 K data were obtained from 23 of 73 middle age men without clinically evident cardiovascular disease who participated in the Aragon Workers Health Study in 2009 (baseline visit) and had available baseline urinary metals and subclinical atherosclerosis measures obtained in 2010–2013 (follow-up visit). The median metal levels were 7.36 µg g⁻¹, 0.33 µg g⁻¹, 0.11 µg g⁻¹ and 0.07 µg g⁻¹, for arsenic (sum of inorganic and methylated species), cadmium, antimony and tungsten, respectively. Urine cadmium and tungsten were associated with femoral and carotid intima-media thickness, respectively (Pearson's <i>r</i> = 0.27; <i>p</i> = 0.03 in both cases). Among nearest genes to identified differentially methylated regions (DMRs), 46% of metal-DMR genes overlapped with atherosclerosis-DMR genes (<i>p</i> < 0.001). Pathway enrichment analysis of atherosclerosis-DMR genes showed a role in inflammatory, metabolic and transport pathways. In <i>in silico</i> protein-to-protein interaction networks among proteins encoded by 162 and 108 genes attributed to atherosclerosis- and metal-DMRs, respectively, with proteins known to have a role in atherosclerosis pathways, we observed hub proteins in the network associated with both atherosclerosis and metal-DMRs (e.g. <i>SMAD3</i> and <i>NOP56</i>), and also hub proteins associated with metal-DMRs only but with relevant connections with atherosclerosis effectors (e.g. <i>SSTR5</i>, <i>HDAC4</i>, <i>AP2A2</i>, <i>CXCL12</i> and <i>SSTR4</i>). Our integrative <i>in silico</i> analysis demonstrates the feasibility of identifying epigenomic regions linked to environmental exposures and potentially involved in relevant pathways for human diseases. While our results support the hypothesis that metal exposures can influence health due to epigenetic changes, larger studies are needed to confirm our pilot results.This article is part of a discussion meeting issue ‘Frontiers in epigenetic chemical biology’