Τα αυξημένα επίπεδα εμβρυϊκής αιμοσφαιρίνης (HbF) μετριάζουν την βαρύτητα των διαταραχών που αφορούν στην β-σφαιρίνη, δηλαδή τη δρεπανοκυτταρική αναιμία (SCD) και την β-μεσογειακή αναιμία, που αποτελούν σημαντικές αιτίες παγκόσμιας νοσηρότητας και θνησιμότητας. Γι’ αυτό το λόγο είναι μακροχρόνιο το ενδιαφέρον για την ανάπτυξη θεραπευτικών προσεγγίσεων για την επαγωγή της παραγωγής HbF. Η αναζήτηση μορίων που ρυθμίζουν την μετάβαση από την έκφραση της εμβρυϊκής (HbF) στην έκφραση της αιμοσφαιρίνης των ενηλίκων (HbA) και που συντελούν στην διατήρηση της αποσιώπησης ή αντίθετα στην ενεργοποίηση της έκφρασης της HbF στους ανθρώπους αποτελεί πολυετές αντικείμενο έρευνας με σκοπό την στόχευση αυτών των παραγόντων για την επαγωγή της HbF (Sankaran et al. 2011). Έτσι, εκτός από τα cis-ρυθμιστικά στοιχεία, έχουν εντοπιστεί και trans-ρυθμιστικά στοιχεία, τα οποία αποτελούν κυρίως μεταγραφικούς παράγοντες. Όμως υπάρχουν και γονιδιακοί τόποι εκτός του β-συμπλέγματος που φαίνεται να επιδούν στην ρύθμιση της έκφρασης των γονιδίων του β-γονιδιακού τόπου. Τέτοιοι είναι οι τόποι που συνδέονται με «ποσοτικά γνωρίσματα» (Quantitative trait loci-QTL). Η χρωμοσωμική περιοχή 6q23 έχει σε διάφορες μελέτες προσδιοριστεί ως QTL, που συνδέεται με την μεταβολή των επιπέδων της HbF σε ασθενείς με SCD. (Close et al. 2004, Thein et al. 2007, Wyszynski et al. 2004). Ο σκοπός της παρούσας μελέτης είναι διττός: A. Ο εντοπισμός μονονουκλεοτιδικών πολυμορφισμών (SNP) εντός των γονιδίων MAP3K5 και PDE7B του QTL στην 6q23 χρωμοσωμική περιοχή, που να σχετίζονται με αυξημένα επίπεδα HbF. B. Η αξιολόγηση των SNP αυτών ως φαρμακογονιδιωματικών δεικτών, που να σχετίζονται με την μεταβλητότητα των επιπέδων της HbF ως απόκριση στη θεραπεία με HU. / Hemoglobinopathies, particularly β-thalassemia and sickle cell disease (SCD), are major health problems, in which quantitative or qualitative defects in hemoglobin production occur, respectively. Under normal circumstances, different types of hemoglobin (Hb) are produced during embryonic, fetal, and adult life. At birth, fetal hemoglobin (HbF), in particular, composes 80–90% of the total hemoglobin synthesized, but it gradually decreases to approximately 1% by 10 months in infancy as its synthesis is restricted to a small subset of erythrocytes termed ‘F cells’ [Patrinos&Grosveld, 2008]. The first studies searching for regulators of HbF expression were conducted on individuals with heterocellular hereditary persistence of HbF (HPFH) – i.e. increase of HbF levels unevenly distributed among ‘F cells’ – and suggested the absence of linkage between the determinant of the HbF levels and the β-globin gene cluster, back then named “non-α globin cluster” [Gianni et al., 1983]. Later, while seeking for genetic elements associated with elevated HbF levels in healthy adults, several cis-acting variants on the β-globin gene complex were unraveled, including the XmnI-Gγ (HBG2) gene promoter polymorphism [Gilman et al., 1985]. Ιn addition, variants unlinked to the β-locus (trans-acting), such as quantitative trait loci (QTLs) on Xp22 [Dover et al., 1992] and 6q23 [Craig et al., 1996] became known soon after. Initially, a study on an extensive, inbred kindred of Asian Indian origin with heterocellular HPFH revealed that a key locus controlling HPFH resides on chromosome 6q, which was fine-mapped to 6q22.3–23.1 [Craig et al., 1996]. Among the first positional candidate genes in the 6q23 region, assumed to possibly explain this QTL, were the MYB proto-oncogene and the eukaryotic release factor-similar HBS1L, as well as the mitogen-activated protein kinase kinase kinase 5 (MAP3K5) [Game et al., 2000]. In addition, genes within this region are associated with response to hydroxyurea (HU) treatment based on elevated HbF levels, in SCD patients; however, the mechanism by which this chromosome 6q22-23 QTL influences HbF levels in the context of HU treatment remains unknown [Ma et al., 2007] and very few, if any, studies have addressed this question. In continuing the global effort of scrutinizing the 6q23 region for variants accounting for the modulation of HbF production, we investigated a possible association of SNPs residing within the MAP3K5 and PDE7B genes with elevated HbF levels in β-thalassemia intermediate or major patients and normal (non-thalassemic) individuals. We also examined a cohort of 38 heterozygous SCD/β-thalassemia patients who had undergone HU therapy, in order to clarify whether there is a correlation of these SNPs with HU treatment response in patients of Hellenic origin.

INTRODUCTION AND AIM - Cardiovascular disease is globally a major risk factor for morbidity and mortality. It is caused by various factors, one of which is an abnormal haemostatic process. Fibrinogen is a haemostatic factor that is considered to be an independent risk factor for cardiovascular disease. Elevated fibrinogen can be caused by environmental and genetic factors which increase the risk of the occurrence of thrombosis. The fibrinogen y' chain, which is one of the three chains of fibrinogen, has two different variants, the yA and y’. The presence of the fibrinogen y’ chain has been associated with thrombotic disorders. Many studies have investigated the fibrinogen variables in Caucasian individuals, but only a few such studies have been conducted on non-Caucasian individuals. The genetic diversity of ethnic groups differs and could cause differences in the fibrinogen variables between these groups. Fibrinogen is known to increase with age; therefore to explain changes over time in fibrinogen concentrations it was also important to investigate whether genetic determinants and possible gene–environment interactions influenced fibrinogen over time. In this study the main aim was to determine the change in the fibrinogen variables over a five-year period within a black South African cohort subdivided according to genotypes associated with fibrinogen variables, and to determine whether the observed changes were modulated by environmental factors. PARTICIPANTS AND METHODS - Data [baseline (n=2010) and follow-up (n=1288)] were collected in the Prospective Urban and Rural Epidemiology (PURE) study during 2005 and 2010 from apparently healthy black men and women aged between 35 and 65 years and residing in rural or urban settlements. Experimental methods included analysis of fibrinogen and fibrinogen y’ concentrations, single nucleotide polymorphisms (SNPs) and determination of environmental factors associated with the fibrinogen variables. RESULTS - The fibrinogen variables increased significantly from 2005 to 2010 in both the rural and urban participants, as well as in both men and women. The major environmental factors that affected the fibrinogen variables were C-reactive protein (CRP), interleukin-6 (IL-6), body mass index (BMI), glycated haemoglobin (HbA1c), age, blood lipids, human immunodeficiency virus (HIV) and tobacco use. Fibrinogen increased consistently from 2005 to 2010 in the respective genotypes of all SNPs analysed, except in the FGG 9340 T>C homozygous mutant carriers. Fibrinogen y’ also increased in general in most genotypes from 2005 to 2010, except in the FGG 10034 C>T mutant allele carriers, where a decrease was observed. It was determined that CRP was the only environmental factor that influenced the change in fibrinogen over time and that FGG 10034 C>T was the only SNP that influenced the change in fibrinogen y’ over the five years. Four gene–environment interactions also influenced fibrinogen on a cross-sectional level, i.e. FGA 2224 G>A with age, FGB Arg448Lys with HIV status, FGB 1643 C>T with urbanisation and FGB 1038 G>A with HbA1c. Only the FGG 9340 T>C with HbA1c interaction was found to predict change in fibrinogen concentrations over the five years. CONCLUSION - Both environmental and genetic factors significantly influenced the fibrinogen variables cross-sectionally as well as prospectively. It was clear that the influence of the environmental factors was mediated by genetic polymorphisms and vice versa, as can be seen by the gene–environment interactions found in this study. An important finding of this study was that the interaction of HbA1c with two SNPs on fibrinogen variables may explain the known inconsistent relationship found between fibrinogen concentrations and diabetes. / MSc (Dietetics), North-West University, Potchefstroom Campus, 2014

Le dihydrofolate réductase (DHFR) est la principale cible du méthotrexate, un important composant du traitement de la leucémie lymphoblastique aiguë (LLA). Une association des polymorphismes du promoteur de DHFR avec l’issue de la LLA a été mise en évidence au laboratoire. Une survie sans événement (EFS) réduite corrélait avec les allèles A -317 et C -1610, et l’haplotype *1, défini par ces allèles. L’haplotype *1 était aussi associé à une expression élevée du DHFR. Dans cette étude, nous étendons l’analyse à la région régulatrice adjacente, d’environ 400 pb, correspondant au transcrit mineur non-codant du DHFR, qui joue un rôle essentiel dans la régulation de la transcription au niveau du promoteur majeur. Six polymorphismes ont été identifiés, parmi lesquels 5 étaient des SNPs et un polymorphisme de longueur composé d’un nombre variable d’éléments de 9 pb et d’une insertion/délétion de 9 pb. L’analyse d’haplotype, incluant tous les polymorphismes promoteurs, a révélé une diversification de l’haploytpe *1 en 5 sous-types (*1a à *1e). Les variations du promoteur majeur et les sous-types de l’haplotype *1 ont été par la suite analysés pour l’association avec l’issue de LLA. Un EFS réduit corrélait avec l’allèle A du polymorphisme G308A (p=0,02) et avec l’haplotype *1 (p=0,01). Des niveaux élevées d’ARNm étaient trouvés chez les porteurs de l’haplotype *1b (p=0,005) et pas pour les autres sous-types de l’haplotype *1. Alors, la mauvaise issue de LLA associée avec l'haplotype *1 est en effet déterminée par le sous-type *1b. Cette étude donne un nouvel aperçu des polymorphismes régulateurs du DHFR définissant plus précisément les variations du DHFR prédisposant un événement. / Dihydrofolate reductase (DHFR) is the major target of methotrexate, a key component in childhood acute lymphoblastic leukemia (ALL) treatment. We recently reported an association of DHFR promoter polymorphisms with ALL outcome. Lower event free survival (EFS) correlated with the alleles A -317 and C -1610, and with haplotype *1, defined by these alleles. Haplotype*1 was also associated higher DHFR expression. Here we extended the analysis to adjacent 400bp regulatory region corresponding to non-coding minor DHFR transcript which plays an essential role in the regulation of transcription from the major promoter. Six polymorphisms were identified, of which 5 were SNPs and one length polymorphism composed of variable number of 9bp elements and 9bp insertion/deletion. Haplotype analysis including all promoter polymorphisms revealed diversification of haplotype *1 into 5 subtypes (*1a to *1e). Major promoter variations and haplotype *1 subtypes were subsequently analyzed for the association with ALL outcome. Lower EFS correlated with an A allele of G308A polymorphism (p=0.02) and with *1b haplotype (p=0.01). Higher mRNA levels were found in the carriers of *1b haplotype (p=0.005) and not for remaining haplotype *1 subtypes. So, the worse ALL outcome associated with haplotype *1 is actually determined by the subtype *1b. The study provides a new insight into DHFR regulatory polymorphisms defining more precisely event–predisposing DHFR variations.

DNA damage response is a complex system responsible for protection of a cell against internal and external DNA damaging agents and in maintaining genome integrity. Many of genes participating in DNA damage response pathways are polymorphic. Genetic polymorphisms in coding and regulatory regions may have impact on the function of proteins encoded by the genes. Phenotypic effect of single nucleotide polymorphisms (SNPs) is subject of investigation in connection with the ability of a cell to manage genotoxic stress and subsequently, in relation to cancer susceptibility. The aim of this thesis was to evaluate the association between SNPs in DNA repair genes (hOGG1, XRCC1, XPC) and cell cycle genes (TP53, p21CDKN1A , BCL2 and BAX) and their mRNA expression in peripheral blood lymphocytes from individuals occupationally exposed to styrene and control individuals. The aim was extended to analyses of relationships between mRNA expression levels of the above-mentioned genes and markers of exposure to styrene (concentration of styrene in blood and in air), markers of DNA damage (single strand breaks - SSBs, and endonuclease III specific sites - Endo III sites) and the base excision repair (BER) capacity, by means of γ-irradiation specific DNA repair rates and oxidative repair. Study on the group of healthy...

Objectif : Ce travail s’inscrit dans le cadre d’une recherche étiologique sur les leucémies aigues (LA) de l’enfant, pathologies dont les facteurs de risque sont peu connus. Plusieurs hypothèses ont été testées : 1) le rôle protecteur d’une supplémentation maternelle en acide folique avant et pendant la grossesse et l’investigation par une approche gène-candidat du rôle de polymorphismes communs rs1801133 et 1801131 de MTHFR et rs1801394 et rs1532268 de MTRR supposés modifier le métabolisme des folates, 2) l’association entre la profession et des expositions professionnelles maternelles lors de la grossesse, 3) l’existence d’un lien positif avec l’exposition des enfants au trafic routier. Matériel et méthodes : Les données analysées proviennent d’une étude cas-témoins française, ESCALE, basée sur le Registre National des Hémopathies malignes de l’Enfant et réalisée en population générale sur la période 2003-2004. L’échantillon comportait 648 cas de leucémie aiguë lymphoblastique [LAL], 116 cas de leucémie aiguë non lymphoblastique [LANL], et 1681 témoins de moins de 15 ans. L’échantillonnage a été stratifié sur l’âge et le sexe. Les données ont été recueillies auprès des mères à l’aide d’un questionnaire téléphonique standardisé, identique pour les cas et les témoins. Les génotypes ont été obtenus par génotypage à haut débit, pangénomique pour les cas et à façon pour les témoins, et par imputation pour les polymorphismes non génotypés. Le géocodage des adresses et des indicateurs dérivés de données d’émission de trafic ont permis d’estimer l’exposition des enfants au trafic routier. Les odds ratios (OR) ont été estimés à l’aide de modèles de régression logistique non conditionnelle, incluant les facteurs de confusion potentiels. Résultats : Le risque de LA était significativement inversement associé à une supplémentation maternelle en acide folique avant ou pendant la grossesse (OR=0.4 [0.3-0.6]). Aucun des polymorphismes génétiques de MTHFR et de MTRR n’était associé au risque de LA. Cependant, le fait d’être à la fois porteur homozygote des allèles variants de l’un des polymorphismes de MTHFR, et porteur de deux allèles variants des polymorphismes de MTRR était positivement associé au risque de LA (OR=1.6 [0.9-3.1]). Nous n’observions pas d’interaction entre MTHFR, MTRR et une supplémentation maternelle en acide folique. Des associations positives et significatives on été mises en évidence entre le risque de LA et des expositions auto-déclarées professionnelles maternelles pendant la grossesse, aux teintures de cheveux (OR=3.0 [1.7-5.2]), à des peintures ou vernis et/ou colles (OR=1.5 [1.1-2.2]), et à des rayonnements ionisants (OR=2.4 [1.3-4.6]). Cependant, ces associations étaient limitées aux fréquences d’exposition de moins d’1 heure par semaine et peuvent refléter une sur-déclaration chez les cas. L’exposition maternelle professionnelle aux pesticides n’était pas associée aux LA dans notre étude.Les LA étaient significativement associées à des concentrations élevées de NO2 de fond liées au trafic estimées au lieu de résidence (OR=1.2 [1.0-1.5]) et avec la présence de routes à fort trafic dans un rayon de 500 m centré sur ce lieu (OR=2.0 [1.0-3.6]). Nous observions une association significative entre les LA et une densité élevée de routes à fort trafic dans un rayon de 500 m (OR=2.2 [1.1-4.2]), avec une tendance linéaire positive significative de l’association des LAL avec la longueur totale de routes à fort trafic dans un rayon de 500 m.Conclusion : Cette thèse apporte des arguments en faveur du rôle protecteur d’une supplémentation maternelle périconceptionnelle en acide folique. Elle suggère également un rôle des polymorphismes de MTHFR et MTRR dans le risque des LA, sans interaction toutefois avec la supplémentation. Enfin, elle renforce l’hypothèse que vivre près de routes à fort trafic pourrait augmenter le risque de LA. / Objectives: This work investigated three hypotheses related to the etiology of childhood acute leukemia (AL):1) maternal folic acid supplementation before or during pregnancy reduces AL risk, accounting for the SNPs rs1801133 (C677T) and rs1801131 (A1298C) in MTHFR and rs1801394 (A66G) and rs1532268 (C524T) in MTRR, assumed to modify folate metabolism, 2) maternal occupation and occupational exposure during pregnancy may be link to LA 3) traffic is a source of environmental exposures, including benzene, which may be related to childhood leukemia. Methods: The data were obtained from the national registry-based case-control study ESCALE, carried out in France in 2003-2004. The ESCALE study included 764 cases and 1681 controls less than 15 years old and the controls were frequency matched with the cases on age and gender. The data were collected by a standardized telephone interview of the mothers. Various indicators of exposure to traffic and pollution were determined using the geocoded addresses at the time of diagnosis for the cases and of interview for the controls. The genotypes were obtained using high-throughput platforms and imputation for untyped polymorphisms. CITP-68 classification was used to classify maternal occupation during pregnancy. Indicators of the distance from, and density of, main roads and background traffic NO2 concentrations data were used. Odds ratio (OR) were estimated using unconditional regression models adjusted for potential confounders. Results: AL was significantly inversely associated with maternal folic acid supplementation before and during pregnancy (OR=0.4 [0.3–0.6]). MTHFR and MTRR genetic polymorphisms were not associated with AL. However, AL was positively associated with homozygosity for any of the MTHFR polymorphisms and carriership of both MTRR variant alleles (OR=1.6 [0.9–3.1]). No interaction was observed between MTHFR, MTRR, and maternal folate supplementation. Significant positive associations were observed between childhood AL and self-reported maternal occupational exposures during pregnancy to hair dye (OR=3.0 [1.7-5.2]), to paints or polishs and/or glues (OR=1.5 [1.1-2.2]), and to ionising radiations (OR=2.4 [1.3-4.6]). However, these associations were limited to exposure frequencies of less than 1 hour by week. Maternal occupational exposure to pesticides during pregnancy was not related to AL. AL was significantly associated with high estimates of traffic NO2 concentration at the place of residence(OR=1.2 [1.0-1.5]) and with the presence of a heavy-traffic road within 500 meters compared to the absence of a heavy-traffic road in the same area (OR=2.0 [1.0-3.6]). There was a significant association between AL and a high density of heavy-traffic roads within 500 meters in comparison to the reference category with no heavy-traffic road within 500 meters (OR=2.2 [1.1-4.2]), with a significant positive linear trend of the association of AL with the total length of heavy-traffic road within 500m. Conclusion: The study findings support the hypothesis that maternal folic acid supplementation may reduce the risk of childhood AL. The findings also suggest that the homozygous genotype for any of the MTHFR variants and carrying both MTRR variants could be a risk factor for AL. Finally, the results support the hypothesis that living close to heavy-traffic roads may increase the risk of childhood leukemia.

A depressão pós-parto (DPP) tem sido associada com alterações no funcionamento do eixo hipotálamo-hipófise-adrenal (HHA), mas pouco se sabe do envolvimento de estruturas cerebrais, ou outros mecanismos subjacentes a estas alterações. Uma hipótese fundamental é que o estresse inerente ao período puerperal, vulnerabilidade individual e, principalmente, as alterações hormonais decorrentes do final da gravidez desempenham um importante papel causal nas alterações do eixo HHA e na incidência da DPP. Estudos sobre o transtorno depressivo maior mostram que alterações funcionais em áreas cerebrais como o giro do cíngulo anterior (GCA) estão relacionadas com humor deprimido, e outros pesquisadores investigaram a relação entre a neuroquímica do GCA e a atividade do eixo HHA. Pesquisas sobre genes de interesse do eixo HHA também têm reportado associações entre polimorfismos nestes genes e alterações nos níveis de cortisol. O presente trabalho testou a hipótese de que mulheres deprimidas no puerpério remoto apresentariam atenuação no funcionamento do eixo HHA, medido pelos níveis de cortisol em 30 minutos após o despertar (CAR) e ao longo da variação diurna (VD); e também que polimorfismos em um gene do eixo HHA, o gene promotor do receptor do tipo 1 do hormônio liberador de corticotrofina (CRHR1), estariam associados com sintomas depressivos no puerpério para prever os níveis de cortisol; e finalmente que as alterações verificadas no funcionamento do eixo HHA de puérperas deprimidas teriam relação com a neuroquímica do GCA. Os resultados indicaram que (1) ao redor do sexto mês após o parto, o CAR e a VD estavam atenuados em puérperas deprimidas comparadas com puérperas eutímicas, e com controles saudáveis não-puérperas; (2) os metabólitos presentes no GCA tinham correlação com as medidas do eixo HHA nas puérperas deprimidas; e (3) a presença de sintomas depressivos em associação com polimorfismos do CRHR1 previram alterações nos níveis de cortisol. No geral, estes resultados sugerem que as alterações do eixo HHA de puérperas deprimidas no puerpério tardio estão associadas com fatores genéticos e com a neuroquímica funcional do GCA / Postpartum depression (PPD) has been associated with changes in the functioning of the hypothalamic-pituitary-adrenal (HPA) axis, but little is known about the involvement of brain structures, or other mechanisms underlying these changes. A key assumption is that stress inherent to the puerperal period, individual vulnerability, and especially the hormonal changes resulting from the end of pregnancy play an important causal role in the alterations of the HPA axis and in the incidence of PPD. Studies on major depressive disorder show that functional changes in brain areas, such as the anterior cingulate gyrus (ACG), are related to depressed mood, and other researchers investigated the relation between the neurochemistry of the ACG and the activity of the HPA axis. Research on the HPA axis genes of interest have also reported associations between polymorphisms in these genes and changes in cortisol levels. The present study tested the hypothesis that depressed women in the remote postpartum period would show attenuation in the functioning of the HPA axis, measured by cortisol levels 30 minutes after awakening (cortisol awakening response, CAR) and by diurnal variation (DV) throughout the day; and also that polymorphisms in a gene of the HPA axis, the promoter gene of the corticotropin releasing hormone receptor type 1 (CRH-R1), would present association with depressive symptoms in the postpartum period to predict the levels of cortisol; and finally that the changes in the functioning of the HPA axis of postpartum depressed women have a relationship with the neurochemistry of the ACG. Results indicated that (1) around the sixth month after delivery, CAR and DV were attenuated in depressed postpartum women compared with euthymic postpartum women and with non-postpartum healthy control women; (2) metabolites present in the ACG showed correlation with measures of the HPA axis in depressed postpartum women; and (3) the presence of depressive symptoms in association with CRHR1 polymorphisms predicted changes in cortisol levels. Overall, these results suggest that changes in the functioning of the HPA axis of depressed postpartum women in the remote postpartum period are associated with genetic factors and with the functional neurochemistry of the ACG

A nefropatia diabética (ND) é uma das principais causas de nefropatia crônica, o que torna o diabetes mellitus (DM) responsável por 44% da prevalência de doença renal crônica (DRC) no mundo. O papel do estresse oxidativo na patogênese da ND está bem estabelecido e genes pertencentes a vias pró- e antioxidantes são possíveis candidatos a conferirem susceptibilidade genética a essa e a outras complicações crônicas. Além do estresse oxidativo, o transporte intracelular de glicose, mediado por transportadores específicos, também parece exercer influência sobre a ND e outras complicações. O objetivo deste trabalho foi avaliar a associação entre ND e alguns polimorfismos de um único nucleotídeo (SNPs) em genes que codificam proteínas transportadoras de glicose (GLUT2 [SLC2A2]), proteínas pró-oxidantes (p22phox [CYBA] e NOX-2 [CYBB]) e proteínas antioxidantes (glutationa peroxidase-4 [GPX4] e catalase [CAT]) em uma coorte brasileira (n=453; 45,8% de pacientes com ND) e três coortes francesas (SURGENE [n=340; 17,7% de pacientes com ND na fase basal], GENEDIAB [n=313; 66,7% de pacientes com ND] e GENESIS [n=636; 49,7% de pacientes com ND]) de pacientes portadores de DM tipo 1. Os SNPs foram genotipados com o uso da técnica de reação em cadeia da polimerase (PCR) em tempo real e os resultados expressos em odds ratio (OR) ou hazard ratio (HR), com seus respectivos intervalos de confiança (IC), determinados em modelos ajustados de regressão logística politômica ou regressão de risco proporcional de Cox, respectivamente. A razão albumina/creatinina urinária (ACR) ou a taxa de excreção urinária de albumina (EUA) foram utilizadas para definir os estágios de ND e os pacientes foram classificados de acordo com a presença ou ausência de ND incipiente (ACR 30 - 300 mg/g de creatinina ou EUA 20 - 200 ?g/min ou 20 - 200 mg/L) e creatinina plasmática <1,7 mg/dL), ND estabilizada (ACR >300 mg/g de creatinina ou EUA > 200 ug/min ou > 200 mg/L e creatinina plasmática < 1,7 mg/dL ) ou ND avançada (ACR > 300 mg/g de creatinina ou EUA > 200 ug/min ou > 200 mg/L e creatinina plasmática > 1,7 mg/dL ou qualquer terapia de reposição renal) e também foram avaliadas associações dos SNPs com a taxa de filtração glomerular estimada (TFGe). O alelo raro A do SNP rs6610650 no gene CYBB foi associado com valores baixos de TFGe em mulheres na coorte brasileira e com a prevalência de ND estabilizada/avançada em mulheres da coorte francesa (OR 1,75; IC 95% 1,11 - 2,78; p=0,016). O alelo raro T do SNP rs713041 no gene GPX4 foi inversamente associado com a prevalência de ND estabilizada/avançada em homens na coorte brasileira (OR 0,30, IC95% 0,13 - 0,68, p=0,004) e com valores elevados de TFGe em homens na coorte francesa. O alelo raro A do SNP rs7947841 no gene CAT foi associado com a prevalência de ND incipiente (OR 2,79; IC95% 1,21 - 6,24; p=0,01) e ND estabilizada/avançada (OR 5,72; IC95% 1,62 - 22,03; p=0,007), bem como com a incidência de eventos renais, definidos como novos casos de microalbuminúria ou progressão para um estágio mais grave de ND durante o seguimento de estudo, na coorte SURGENE (HR 1,82; IC95% 1,13 - 2,81; p=0,01). O mesmo alelo de risco associou-se com a prevalência de ND incipiente (OR 3,13; IC95% 1,42 - 7,24; p=0,004) e com a incidência de insuficiência renal crônica terminal (IRCT) na coorte GENEDIAB (HR 2,11; IC95% 1,23 - 3,60; p=0,008) e com a prevalência de ND incipiente (OR 2,16; IC95% 1,14 - 4,10, p=0,02) e ND estabilizada/avançada (OR 2,71; IC95% 1,38 - 5,42; p=0,004) na coorte brasileira. O alelo raro T do SNP rs9932581 no gene CYBA foi inversamente associado com a prevalência de ND estabilizada/avançada (OR 0,60; IC95% 0,46 - 0,78; p=0,0001) e com valores mais baixos de TFGe nos pacientes de descendência europeia da coorte GENESIS/GENEDIAB. Este mesmo alelo foi associado com a incidência de eventos renais e de IRCT nas coortes SURGENE (HR 0,63; IC95% 0,46 - 0,86; p=0,003) e GENESIS/GENEDIAB (HR 0,51; IC95% 0,31 - 0,78; p=0,002), respectivamente. Entretanto estes resultados não foram replicados na coorte brasileira. O alelo raro T do SNP rs11924032 no gene SLC2A2 foi inversamente associado com a perda da TFGe ao logo do tempo (0,02%/ano vs 2,18%/ano para os pacientes portadores do genótipo GG; p=0,005), na coorte SURGENE. Este mesmo alelo foi inversamente associado com a incidência de IRCT nas coortes GENESIS/GENEDIAB (HR 0,53; IC95% 0,29 - 0,89; p=0,01). Os resultados observados para o gene SLC2A2 não forneceram fortes indícios para afirmarmos que este gene exerça um papel relevante no desenvolvimento da ND nos pacientes com DM tipo 1 nas coortes francesas estudadas. Em contrapartida, os SNPs nos genes que codificam as proteínas pró-oxidantes CYBA e CYBB e as proteínas antioxidantes GPX-4 e CAT foram capazes de modular o risco para doença renal em pacientes portadores de DM tipo 1, sendo que os SNPs presentes nos genes CYBB, GPX4 e CAT tiveram seus resultados replicados em coortes independentes, o que corrobora a importância destes genes e, consequentemente, do estresse oxidativo, na patogênese da ND / Diabetic nephropathy (DN) is a major cause of chronic nephropathy, with diabetes mellitus (DM) accounting for 44% of the prevalence of chronic kidney disease (CKD) in the world. The role of oxidative stress in the pathogenesis of DN is well established and genes belonging to pro- and antioxidant pathways are possible candidates to confer genetic susceptibility to this and other chronic complications. Besides oxidative stress, intracellular glucose transport mediated by specific transporters, also appears to influence DN and other complications. The aim of this study was to evaluate the association between DN and some single nucleotide polymorphisms (SNPs) present in genes encoding glucose transport proteins (GLUT2 [SLC2A2]), pro- (p22phox [CYBA] and NOX-2 [CYBB]) and antioxidants (glutathione peroxidase-4 [GPX4] and catalase [CAT]) proteins, in a Brazilian cohort [n= 453; 45.8% f patients with DN], and three French cohorts (SURGENE [n=340; 17.7% of patients with DN at baseline], GENEDIAB [n=313; 66.7% of patients with DN], and GENESIS [n=636; 49.7% of patients with DN]) of patients with type 1 DM. The SNPs were genotyped using the technique of real time polymerase chain reaction (PCR) and results expressed as odds ratio (OR) and hazard ratio (HR), with their respectively 95% confidence intervals (CI), determined by adjusted models of polytomic logistic regression and Cox proportional hazard regression, respectively. The albumin/creatinine ratio (ACR) or the urinary albumin excretion (UAE) rate were used to define the DN stages and the patients were classified according to the presence or absence of incipient DN (ACR 30 - 300 mg/g of creatinine or UAE 20 - 200 ug/min or 20 - 200 mg/L) and plasmatic creatinine < 1,7 mg/dL), established DN (ACR > 300 mg/g of creatinine or EUA > 200 ug/min or > 200 mg/L and plasmatic creatinine <1,7 mg/dL) or advanced DN (ACR >300 mg/g of creatinine or UAE > 200 ug/min or > 200 mg/L and plasmatic creatinine > 1,7 mg/dL or any renal replacement therapy). Associations for the estimated glomerular filtration rate (eGFR) were also evaluated. The rare allele A of the SNP rs6610650 in CYBB gene was associated with low values of eGFR in women in the Brazilian cohort and with the prevalence of established/advanced DN in women in the French cohort (OR 1.75, 95%CI 1.11 - 2.78, p=0.016). The rare allele T of the SNP rs713041 in GPX4 gene was inversely associated with the prevalence of established/advanced DN in men in the Brazilian cohort (OR 0.30, 95%CI 0.13 - 0.68, p=0.004) and with higher values of eGFR in men in the French cohort. The rare allele A of the SNP rs7947841 in CAT gene was associated with the prevalence of incipient DN (OR 2.79, 95%CI 1.21 - 6.24, p=0.01) and established/advanced DN (OR 5.72; 95%CI 1.62 - 22.03, p=0.007) as well as the incidence of renal events, defined as new cases of microalbuminuria or progression to a more severe stage during the follow-up study, in SURGENE cohort (HR 1.82, 95%CI 1.13 - 2.81, p=0.01). The same risk allele was associated with the prevalence of incipient DN (OR 3.13, 95%CI 1.42 - 7.24, p=0.004), the incidence of end-stage renal disease (ESRD) in the cohort GENEDIAB (HR 2.11, 95%CI 1.23 - 3.60, p=0.008) and with the prevalence of incipient DN (OR 2.16, 95%CI 1.14 - 4.10, p=0.02) and established/advanced DN (OR 2.71, 95%CI 1.38 - 5.42, p=0.004) in the Brazilian cohort. The rare T allele of the SNP rs9932581 in CYBA gene was inversely associated with the prevalence of established/advanced DN (OR: 0.60, 95%CI: 0.46 - .78, p=0.0001) and associated with lower values of eGFR in patients of GENESIS/GENEDIAB cohort. The same allele was inversely associated with the incidence of renal events and ESRD in SURGENE (HR 0.63, 95%CI 0.46 - 0.86, p=0.003) and GENESIS/GENEDIAB (HR 0.51, 95%CI 0.31 - 0.78, p=0.002) cohorts. However, these results were not replicated in the Brazilian cohort. The rare T allele of the SNP rs11924032 in SLC2A2 gene was inversely associated with the loss of eGFR during the follow-up (0.02%/year vs. 2.18%/year for patients with the GG genotype, p=0.005) in the SURGENE cohort. The same allele was inversely associated with the incidence of ESRD in the GENESIS/GENEDIAB cohorts (HR 0.53, 95%CI 0.29 - 0.89, p=0.01). The results observed for the SLC2A2 gene, in this study, did not provide strong evidence to state that this gene exerts a relevant role in the development of DN in patients with type 1 DM in the studied cohorts. However, SNPs in genes encoding the pro-oxidant proteins CYBA and CYBB, and the antioxidants proteins GPX-4 and CAT were able to modulate the risk of renal disease in patients with type 1 DM. The studied SNPs in CYBB, GPX4 and CAT genes had their results replicated in independent cohorts, which confirms the importance of these genes and, hence, of the oxidative stress in the pathogenesis of DN

INTRODUÇÃO: acredita-se que o controle glicêmico inadequado, a duração do diabetes melito (DM) e a presença de hipertensão arterial e dislipidemia sejam os fatores de risco mais importantes para o desenvolvimento das complicações microvasculares no DM, contudo, existem inúmeras evidências sugerindo que uma predisposição genética participe da suscetibilidade para o desenvolvimento dessas complicações. Vários genes relacionados aos mecanismos dos danos induzidos pela hiperglicemia têm sido investigados. O papel do estresse oxidativo na patogênese das complicações crônicas do DM vem sendo demonstrado e os genes que codificam enzimas que participam dos mecanismos antioxidantes são candidatos a conferirem suscetibilidade ou proteção contra as complicações crônicas. A glutationa é um dos mais importantes antioxidantes endógenos; no entanto, a associação entre polimorfismos em genes que codificam enzimas que participam desse sistema e complicações crônicas do DM foi pouco explorada na literatura. OBJETIVOS: avaliar a associação de polimorfismos em três genes que codificam enzimas relacionadas ao metabolismo da glutationa com o desenvolvimento de nefropatia e retinopatia em pacientes diabéticos tipo 1. Foram estudados: o polimorfismo -129C/T do gene GCLC, o número de repetições do trinucleotídeo GCG no exon 1 do gene GPX1 e o polimorfismo -65T/C do gene GPX3. CASUÍSTICA E MÉTODOS: 299 pacientes (139 do gênero masculino e 160 do gênero feminino) com DM tipo 1 com mais de 15 anos de diagnóstico e mau controle glicêmico foram divididos conforme presença ou ausência das seguintes complicações: nefropatia diabética (ND) avançada, ND, doença renal crônica (DRC) estágios 3 a 5 e retinopatia diabética proliferativa (RDP). Em cada grupo foram avaliadas as freqüências das variantes alélicas dos três genes estudados. RESULTADOS: a distribuição dos genótipos na população estudada foi consistente com o equilíbrio de Hardy-Weinberg para os três genes analisados. A presença de pelo menos um alelo T do polimorfismo -129C/T do gene GCLC conferiu risco independente para a presença de ND avançada (OR = 2,82 ; IC 95% = 1,13 - 7,05; p = 0,026), para ND (OR = 3,64; IC 95% = 1,27 10,36; p = 0,016) e para DRC estágios 3 a 5 (OR = 5,74; IC 95% = 2,17 15,1; p < 0,001) e a presença de pelo menos um alelo C do polimorfismo -65 T/C conferiu risco independente para a presença de ND avançada (OR = 2,62; IC 95% = 1,19 -5,72, p = 0,022) na população estudada. Não houve associação do número de repetições do trinucleotídeo GCG do gene GPX1 com nenhuma das complicações estudadas. O haplótipo CC_TT, composto pelos alelos selvagens dos genes GCLC e GPX3, foi negativamente associado com ND avançada (OR = 0,32, IC 95% = 0,15 0,66; p = 0,002) e DRC (OR = 0,25; IC 95% = 0,11 - 0,55; p = 0,001). CONCLUSÕES: a presença de pelo menos um alelo T do polimorfismo -129 C/T do gene GCLC e de pelo menos um alelo C do polimorfismo -65 T/C do gene GPX3, ambos associados a uma menor atividade transcricional do respectivo gene, conferiram risco para a presença de complicações renais na população de pacientes estudada. / INTRODUCTION: glycemic control, diabetes duration, systemic hypertension and dyslipidemia have been implicated as main risk factors for the development of diabetic microangiopathy, however there is evidence suggesting that genetic predisposition plays a role in the susceptibility to microvascular complications. Based on underlying pathogenesis, polymorphisms of several genes belonging to multiple pathways have been investigated, like the genes related to mechanisms of hyperglycemia-induced damage. The role of oxidative stress in the pathogenesis of diabetic complication has been increasingly demonstrated and genes coding enzymes involved in antioxidant defense are candidates to confer susceptibility or protection against these complications. Glutathione is one the most important endogen antioxidants, however, the association between polymorphisms in genes related to glutathione metabolism and diabetic complications has not been deeply investigated. OBJECTIVES: to study the association between polymorphisms in three genes which code enzymes related to glutathione metabolism and the development of nephropathy and retinopathy in type 1 diabetic patients: the polymorphism -129 C/T of GCLC, the number of trinucleotide GCG repeats at exon 1 of GPX1 and the polymorphism -65 T/C of GPX3. CASUISTIC AND METHODS: 299 type 1 diabetic patients (139 male and 160 female) with at least 15 years from diagnosis and poor glycemic control were studied. The patients were divided in two groups according to the presence or absence of diabetic complications: with and without diabetic nephropathy (DN), advanced DN, chronic kidney disease (CKD) stages 3 to 5 and proliferative diabetic retinopathy (PDR). RESULTS: The allelic distribution of the three studied polymorphisms was consistent with Hardy-Weinberg equilibrium. The presence of at least one T allele of GCLC 129 C/T was an independent risk factor for advanced DN (OR = 2.82 ; CI 95% = 1.13 -7.05; p = 0.026), for DN (OR = 3.64; CI 95% = 1.27 10.36; p = 0.016) and for CKD stages 3 to 5 (OR = 5.74; CI 95% = 2.17 15.1; p < 0.001) and the presence of at least one C allele of GPX3 -65 T/C was an independent risk factor for advanced DN (OR = 2.62; IC 95% = 1.19 -5.72, p = 0.022) in the studied population. There were no associations between GCG trinucleotide repeats of GPX1 and diabetic complications. The haplotype CC_TT, composed by GCLC and GPX3 wild type alleles, was negatively related to advanced DN (OR = 0.32, CI 95% = 0.15 0.66; p = 0.002) and CKD (OR = 0.25; CI 95% = 0.11 0.55; p = 0.001). CONCLUSIONS: the presence of at least one T allele of -129C/T polymorphism of GCLC and one C allele of -65 T/C polymorphism of GPX3, both associated to a lower transcriptional activity of its genes, conferred risk for renal complications in the studied population.

O uso do ácido acetilsalicílico, universalmente aceito na prevenção e tratamento da doença aterosclerótica, pode resultar em respostas terapêuticas variáveis classificando os pacientes em respondedores (normais) ou baixo respondedores (resistentes). A dose de 100mg/dia pode inibir de forma insuficiente a agregação plaquetária. Por isso, doses maiores têm sido utilizadas ou, eventualmente, associadas a outros antiplaquetários visando à redução do número de baixo respondedores. O objetivo do presente estudo foi avaliar os efeitos do ácido acetilsalicílico na redução da agregação plaquetária in vitro. Para tal, 152 indivíduos de ambos os sexos, não aparentados, de qualquer etnia, sem limite de faixa etária, portadores de doença arterial coronária crônica, atendidos no Ambulatório de Coronariopatias do Instituto Dante Pazzanese de Cardiologia, foram incluídos no estudo. Na primeira consulta, todos foram submetidos à anamnese e exame clínico completo e estavam em uso prévio de ácido acetilsalicílico 100mg/dia, por um período superior a 30 dias. Os pacientes selecionados foram divididos em dois grupos: grupo AAS 200, composto por aqueles que foram tratados com ácido acetilsalicílico na dose de 200mg/dia e grupo Ômega 3 composto por aqueles tratados com ácido acetilsalicílico 100mg/dia em associação a ácido graxo ômega 3 na dose de 1.000mg/dia. A agregação plaquetária foi mensurada, in vitro, pelo agregômetro Multiplate, na primeira e segunda consulta. Adicionalmente, amostras de sangue foram obtidas para análise bioquímica e identificação dos polimorfismos nos genes COX-1 (rs 384787; rs 3842798) relacionado com a atividade do ácido acetilsalicílico e ITGB 3 relacionado com a codificação da glicoproteína IIIa (GPIIIa-rs 5918), subunidade do receptor de fibrogênio. Análise da função plaquetária (Aspitest) foi realizada em ambos os grupos. Os valores de corte para o Aspitest, relativo à medida de inibição da ciclooxigenase 1 pelo ácido acetilsalicílico, foram: <= 30 AUC para os muito bons respondedores, > 30 e 40 AUC para os não respondedores. Para a dosagem plasmática de tromboxane, utilizou-se o kit Elisa e, para sua qualificação, a espectrofotometria. Para o DNA genômico, extraído do sangue total periférico, utilizou-se o sistema automatizado QIA cube seguido pela amplificação, pela PCR, da região do DNA que continha o polimorfismo. Os dados foram analisados pelo software SPSS-20 e o nível de significância adotado de 5% (p < 0,05). O teste paramétrico t Student foi utilizado para os dados com distribuição normal (teste Kolmogorov-Smirnov) e os testes não paramétricos Mann-Whitney ou Wilcoxon para os demais dados. A frequência das variáveis qualitativas foi determinada pelo teste do qui-quadrado. A redução dos níveis séricos de VLDL e triglicérides foram semelhantes nos dois grupos e a redução do número de monócidos foi estatisticamente maior no grupo ômega 3. Dos 152 pacientes incluídos no estudo, 38 (25,2%) não eram respondedores ao tratamento prévio com o ácido acetilsalicílico, na posologia de 100mg/dia. A frequência genotípica e alélica dos polimorfismos e a presença do alelo raro foram semelhantes nos grupos respondedor e não respondedor ao ácido acetilsalicílico. A função plaquetária e a produção de tromboxane foram semelhantes nos grupos AAS 200 e Ômega 3. A redução do Aspitest foi observada apenas no grupo não respondedor. A presença do alelo raro do polimorfismo rs 3842787 (gene PTGS1) associou-se à pior resposta do Aspitest e o alelo raro do polimorfismo rs 5918 (gene ITGB3) associou-se à pior resposta à concentração de tromboxane, após 30 dias de tratamento. Em conclusão, 1) os resultados desse estudo mostraram que a associação do ácido acetilsalicílico na dose de 100mg/dia e ômega3 na dose de 1.000mg/dia não reduziu a agregação plaquetária in vitro; 2) os pacientes não respondedores que fizeram uso de ácido acetilsalicílico na dose de 200mg/dia, após 30 dias, tiveram redução no Aspitest e passaram a ser considerados respondedores; 3) A presença dos alelos rs 384787 (COX1-PTGS1) foi responsável pela pior resposta ao Aspitest e a do alelo rs 5918 pela pior resposta ao tromboxane B2. A farmacogenética abre novas perspectivas para o tratamento clínico personalizado da antiagregação plaquetária. / The use of acetylsalicylic acid for the prevention and treatment of atherosclerotic disease may result in different therapeutic responses. Based on that, patients are classified in responders (normal) or low responders (resistant). In clinical practice, the use of 100mg/daily of acetylsalicylic acid may be insufficient for platelet aggregation inhibition, therefore either higher doses or combination with other antiplatelet agents have been used in order to reduce the rates of low responders. The aim of the present study was to evaluate the effects of acetylsalicylic acid in reducing platelet aggregation in vitro. One hundred, fifty-two subjects of both genders, unrelated, of any ethnicity, at any age, and with diagnosis of chronic coronary artery disease followed at the Coronary Artery Disease Section of Dante Pazzanese Institute of Cardiology were included in the study. All patients underwent anamnesis and clinical examination at first consultation. All subjects were on aspirin use (100mg/daily) for at least 30 days before inclusion. Patients were divided into two groups: group ASA, composed by those treated with 200mg of acetylsalicylic acid only and group Omega 3, composed by those treated with 100mg of acetylsalicylic acid in addition to 1.000 mg of omega 3 fatty acid. Platelet aggregation was measured by Multiplate aggregometer at first and second visits. In each visit, blood samples were obtained for biochemical analysis and identification of gene polymorphisms of COX-1 (RS 384 787; rs 3842798) related to the activity of acetylsalicylic acid and of 3 ITGB related glycoprotein IIIa (GPIIIa RS-5918) coding. Platelet function was also analyzed. Cut-off values for Aspitest related to inhibition of cyclooxygenase 1 by acetylsalicylic acid were <= 30 AUC for very good responders, > 30 <= 40 AUC for good responders, and > 40 AUC for non-responders. Elisa test was used for thromboxane plasmatic dosage assessment whereas spectrophotometry was used for its quality evaluation. For genomic DNA extracted from peripheral whole blood, we used QIA cube automated system followed by the amplification by PCR of the region of DNA containing the polymorphism. Data was analyzed by SPSS-20 software. P values < 0.05 were considered statistically significant. Parametric Student t test was used for data with normal distribution (Kolmogorov-Smirnov test) and non-parametric Mann-Whitney or Wilcoxon for other data. The frequency of qualitative variables was analyzed using chi-square test. There was a reduction of VLDL and triglycerides serum levels in both groups, however, the reduction of monocytes was statistically higher in Omega 3 group. Out of 152 patients included in the study, 38 (25.2%) were non responders to prior treatment with acetylsalicylic acid (100mg/daily). Genotypic and allelic polymorphism frequencies and the presence of the rare allele were similar in the responder and non-responder groups. Platelet function and thromboxane production were similar between groups ASA and Ômega 3. Aspitest reduction was observed only in the non-responder group. The presence of rare allele of rs 3842787 polymorphism (PTGS1 gene) was associated with worse response to Aspitest whereas the presence of rare allele of rs 5918 polymorphism (ITGB3 gene) was associated with poor response to thromboxane concentration. In conclusion, 1) the combination of aspirin 100mg/daily and omega 3 1.000 mg/daily did not reduce platelet aggregation in vitro; 2) Non-responders who received aspirin 200mg/daily presented reduction in the Aspitest and were considered responsive after 30 days of treatment; 3) The presence of the alleles rs 384,787 (COX1-PTGS1) was associated with worse response to Aspitest and the presence of the allele rs 5918 was associated with worst response to thromboxane B2. More data is needed to confirm our results. The pharmacogenetics will be an important tool for clinicians in order to customize specific treatment for each patient in platelet aggregation.

A esclerose múltipla (EM), doença neurodegenerativa do sistema nervoso central (SNC) com característica auto-imune e inflamatória, com eventos iniciais, bem como a evolução da EM. É uma doença heterogênea (três principais formas clínicas) e multifatoriais. A imunidade inata demonstrou recentemente ser um fator importante na EM e as variantes genéticas dos componentes do inflamassoma têm sido associadas a doenças autoimunes e neurodegenerativas, com isso hipotetizamos que o inflamassoma e suas citocinas IL-1Beta e IL-18, podem representar importantes contribuintes na patogênese da EM e eventualmente explicar, pelo menos em parte, a heterogeneidade observada em pacientes com EM. Fizemos uma análise multivariada que foi realizada com base na forma clínica (recorrente remitente/RR, primária progressivo /PP ou secundário progressiva /SP, índice de gravidade (EDSS) e índice de progressão (IP). Os monócitos do sangue periférico (PBMC) dos pacientes foram examinados para ativação do inflamassoma (Produção de IL-1Beta e IL-18, clivagem de caspase-1). Com os objetivos de avaliar a contribuição do inflamassoma na EM, em termos de (a) efeito genético sobre o desenvolvimento, gravidade e / ou prognóstico, e (b) ativação complexa de células de sangue periférico como uma forma de avaliar a inflamação sistêmica. Para isso, utilizamos variantes genéticas funcionais em componentes do inflamassoma, que foram analisadas em uma coorte de pacientes com EM, pelo uso de ensaios específicos de alelos e qPCR. A analise multivariada resultou em associação com a variante -511C / T IL1B ganho de função, sendo essa mais frequente em formas progressivas (especialmente SP) do que em RR. A variante de ganho de função NLRP3 Q705K resultou mais frequente em pacientes com EDSS > 3 do que em pacientes com EDSS < 3 e, consequentemente, esse SNP está associado a um IP mais elevado. A análise de PBMC mostrou que as células de indivíduos EM, são mais propensas a responder a um estímulo NLRP3 clássico (isto é, LPS) do que as dos doadores saudáveis. Em conjunto, esses achados indicaram que os pacientes com EM apresentam uma desregulação no inflamassoma NLRP3, podendo ser avaliada no sangue periférico facilitando um prognóstico, e que esse perfil pode ser secundário a um mecanismo genético pró-inflamassoma / The multiple sclerosis (MS), neurodegenerative disease of the central nervous system (CNS) with autoimmune and inflammatory characteristics, with initial events, as well as the evolution of MS, are heterogeneous (three main clinical forms) and multifactorial. Innate immunity has recently been shown to be an important factor in MS and the genetic variants of the components of inflammassoma have been associated with autoimmune and neurodegenerative diseases, thereby hypothesizing that the inflammassoma and its IL-1Beta and IL-18 cytokines may represent important contributors in the pathogenesis of MS and possibly explain, at least in part, the heterogeneity observed in MS patients. We performed a multivariate analysis that was performed based on clinical form (recurrent recurrent / RR, progressive primary / PP or progressive secondary / SP, EDSS and progression index.) Peripheral blood mononuclear cells (PBMC) of patients were examined for inflammatory activation (IL-1Beta and IL-18 production, caspase-1 cleavage). With the objectives of evaluating the contribution of inflammassoma in MS in terms of (a) genetic effect on development, severity and / or prognosis, and (b) complex activation of peripheral blood cells as a way of assessing systemic inflammation. For this, we used functional genetic variants in components of the inflammassoma, which were analyzed in a cohort of MS patients, through the use of specific allele and qPCR assays. For this, we used functional genetic variants in components of the inflammassoma, which were analyzed in a cohort of MS patients, through the use of specific allele and qPCR assays. Multivariate analysis resulted in association with the -511C / T IL1B function gain, which is more frequent in progressive forms (especially SP) than in RR. The gain variant of NLRP3 Q705K function was more frequent in patients with EDSS > 3 than in patients with EDSS < 3 and, consequently, this SNP is associated with a higher PI. PBMC analysis showed that cells from MS individuals are more likely to respond to a classical NLRP3 (ie LPS) stimulus than healthy donors. Taken together, these findings indicated that patients with MS have a dysregulation in the NLRP3 inflammassoma and can be evaluated in the peripheral blood facilitating a prognosis and that this profile may be secondary to a pro-inflammatory genetic mechanism