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BACKGROUND: In Colombia and worldwide, breast cancer (BC) is the most frequently diagnosed neoplasia and the leading cause of death from cancer among women. Studies predominantly involve hereditary and familial cases, demonstrating a gap in the literature regarding the identification of germline mutations in unselected patients from Latin-America. Identification of pathogenic/likely pathogenic (P/LP) variants is important for shaping national genetic analysis policies, genetic counseling, and early detection strategies. The present study included 400 women with unselected breast cancer (BC), in whom we analyzed ten genes, using Whole Exome Sequencing (WES), know to confer risk for BC, with the aim of determining the genomic profile of previously unreported P/LP variants in the affected population. Additionally, Multiplex Ligation-dependent Probe Amplification (MLPA) was performed to identify Large Genomic Rearrangements (LGRs) in the BRCA1/2 genes. To ascertain the functional impact of a recurrent intronic variant (ATM c.5496 + 2_5496 + 5delTAAG), a minigene assay was conducted. RESULTS: We ascertained the frequency of P/LP germline variants in BRCA2 (2.5%), ATM (1.25%), BRCA1 (0.75%), PALB2 (0.50%), CHEK2 (0.50%), BARD1 (0.25%), and RAD51D (0.25%) genes in the population of study. P/LP variants account for 6% of the total population analyzed. No LGRs were detected in our study. We identified 1.75% of recurrent variants in BRCA2 and ATM genes. One of them corresponds to the ATM c.5496 + 2_5496 + 5delTAAG. Functional validation of this variant demonstrated a splicing alteration probably modifying the Pincer domain and subsequent protein structure. CONCLUSION: This study described for the first time the genomic profile of ten risk genes in Colombian women with unselected BC. Our findings underscore the significance of population-based research, advocating the consideration of molecular testing in all women with cancer.
Assuntos
Proteína BRCA2 , Neoplasias da Mama , Predisposição Genética para Doença , Mutação em Linhagem Germinativa , Humanos , Feminino , Mutação em Linhagem Germinativa/genética , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Neoplasias da Mama/epidemiologia , Colômbia/epidemiologia , Pessoa de Meia-Idade , Adulto , Proteína BRCA2/genética , Proteína BRCA1/genética , Sequenciamento do Exoma , Idoso , Testes Genéticos/métodos , Proteínas Mutadas de Ataxia Telangiectasia/genéticaRESUMO
Purpose: Breast Cancer (BC) is the main female cancer diagnosed worldwide, and it has been described that few genes, such as BRCA1, have a high penetrance for this type of cancer. In this manuscript, we were interested in evaluating the effect of 3'UTR variants on BRCA1 expression. Patients and Methods: To accomplish this objective, Whole Exome Sequencing (WES) data of 400 patients with unselected BC was used to filter variants located in the region of interest of BRCA1 gene, finding two of them (c.*36C>G and c.*369_373del). miRGate and miRanda in silico tools were used to predict microRNA (miRNA) interaction. Results: The two variants (c.*36C>G, c.*369_373del) were predicted to affect miRNA interaction. After cloning of BRCA1 3'UTR into pMIR-Report vector, the construct was transfected into two BC cell lines (MDA-MB-231 and MCF-7), and the variant c.*36C>G evidenced overexpression of reporter gene luciferase, showing that the transcript was not being degraded by the miRNA in MDA-MB-231 cells. Conclusion: The variant seems to protect against Triple Negative BC probably due to the expression level of miRNA in this particular cell line (MDA-MB-231). This is consistent with the clinical history of the patients who harbor BC Hormone Receptors positive (HR+).
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Congenital long QT syndrome (LQTS) is a cardiac channelopathy characterized by a prolongation of the QT interval and T-wave abnormalities, caused, in most cases, by mutations in KCNQ1, KCNH2, and SCN5A. Although the predominant pattern of LQTS inheritance is autosomal dominant, compound heterozygous mutations in genes encoding potassium channels have been reported, often with early disease onset and more severe phenotypes. Since the molecular mechanisms underlying severe phenotypes in carriers of compound heterozygous mutations are unknown, it is possible that these compound mutations lead to synergistic or additive alterations to channel structure and function. In this study, all-atom molecular dynamic simulations of KCNQ1 and hERG channels were carried out, including wild-type and channels with compound mutations found in two patients with severe LQTS phenotypes and limited family history of the disease. Because channels can likely incorporate different subunit combinations from different alleles, there are multiple possible configurations of ion channels in LQTS patients. This analysis allowed us to establish the structural impact of different configurations of mutant channels in the activated/open state. Our data suggest that channels with these mutations show moderate changes in folding energy (in most cases of stabilizing character) and changes in channel mobility and volume, differentiating them from each other and from WT. This would indicate possible alterations in K+ ion flow. Hetero-tetrameric mutant channels showed intermediate structural and volume alterations vis-à-vis homo-tetrameric channels. These findings support the hypothesis that hetero-tetrameric channels in patients with compound heterozygous mutations do not necessarily lead to synergistic structural alterations.
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Canalopatias/genética , Canal de Potássio ERG1/metabolismo , Canal de Potássio KCNQ1/metabolismo , Síndrome do QT Longo/genética , Simulação de Dinâmica Molecular , Criança , Pré-Escolar , Canal de Potássio ERG1/genética , Humanos , Canal de Potássio KCNQ1/genética , MasculinoRESUMO
This study aimed at determining the effect of kinesio-taping (KT) on muscle performance and delayed onset muscle soreness (DOMS) after exercise induced muscle damaged. Sixty-six healthy men volunteered to participate (age:18-25 y/o), who performed 200 isokinetic lengthening contractions of the dominant quadriceps. Then subjects were randomized to either control (no treatment), sham (no tape tension), or KT (10% tape tension) groups. Muscle performance was assessed by peak torque and muscular work during maximal isometric and concentric isokinetic contractions. DOMS intensity was assessed using a visual analog scale. Measurements were taken pre-exercise (Pre), 48 h and 96 h post-exercise. Repeated measures ANOVA was used for comparisons within group, and ANCOVA for comparisons among groups. Muscle damage was confirmed in all participants by an increase in CK activity level (p<0.01). Decrease in isometric and isokinetic peak torque was detected at 48 h in the control and sham groups (p<0.01). Muscular work decreased in all groups at 48 h (p<0.01). No differences between groups were detected in muscular performance variables. Increase in DOMS intensity was determined in all groups at 48 h. Comparisons between groups showed lower DOMS intensity in the KT group at 48 h. KT decreased DOMS intensity perception after exercise-induced muscle damage; however, it did not impact muscular performance.
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Desempenho Atlético/fisiologia , Fita Atlética , Mialgia/prevenção & controle , Músculo Quadríceps/fisiologia , Adolescente , Adulto , Creatina Quinase/sangue , Exercício Físico/fisiologia , Humanos , Contração Isométrica/fisiologia , Masculino , Contração Muscular/fisiologia , Percepção da Dor/fisiologia , Músculo Quadríceps/lesões , Músculo Quadríceps/metabolismo , Fatores de Tempo , Torque , Adulto JovemRESUMO
Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are rare severe cutaneous adverse reactions to drugs. Granulysin (GNLY) plays a key role in keratinocyte apoptosis during SJS/TEN pathophysiology. To determine if GNLY-encoding mutations might be related to the protein's functional disturbances, contributing to SJS/TEN pathogenesis, we performed direct sequencing of GNLY's coding region in a group of 19 Colombian SJS/TEN patients. A GNLY genetic screening was implemented in a group of 249 healthy individuals. We identified the c.11G > A heterozygous sequence variant in a TEN case, which creates a premature termination codon (PTC) (p.Trp4Ter). We show that a mutant protein is synthesised, possibly due to a PTC-readthrough mechanism. Functional assays demonstrated that the mutant protein was abnormally located in the nuclear compartment, potentially leading to a toxic effect. Our results argue in favour of GNLY non-synonymous sequence variants contributing to SJS/TEN pathophysiology, thereby constituting a promising, clinically useful molecular biomarker.
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Antígenos de Diferenciação de Linfócitos T/genética , Queratinócitos/patologia , Proteínas Mutantes/metabolismo , Mutação , Necrose , Síndrome de Stevens-Johnson/genética , Síndrome de Stevens-Johnson/patologia , Adolescente , Adulto , Apoptose , Biomarcadores/metabolismo , Estudos de Casos e Controles , Criança , Pré-Escolar , Feminino , Predisposição Genética para Doença , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Proteínas Mutantes/genética , Adulto JovemRESUMO
BACKGROUND: Human reproductive disorders consist of frequently occurring dysfunctions including a broad range of phenotypes affecting fertility and women's health during pregnancy. Several female-related diseases have been associated with hypofertility/infertility phenotypes, such as recurrent pregnancy loss (RPL). Other occurring diseases may be life-threatening for the mother and foetus, such as preeclampsia (PE) and intra-uterine growth restriction (IUGR). FOXD1 was defined as a major molecule involved in embryo implantation in mice and humans by regulating endometrial/placental genes. FOXD1 mutations in human species have been functionally linked to RPL's origin. METHODS: FOXD1 gene mutation screening, in 158 patients affected by PE, IUGR, RPL and repeated implantation failure (RIF), by direct sequencing and bioinformatics analysis. Plasmid constructs including FOXD1 mutations were used to perform in vitro gene reporter assays. RESULTS: Nine non-synonymous sequence variants were identified. Functional experiments revealed that p.His267Tyr and p.Arg57del led to disturbances of promoter transcriptional activity (C3 and PlGF genes). The FOXD1 p.Ala356Gly and p.Ile364Met deleterious mutations (previously found in RPL patients) have been identified in the present work in women suffering PE and IUGR. CONCLUSIONS: Our results argue in favour of FOXD1 mutations' central role in RPL, RIF, IUGR and PE pathogenesis via C3 and PlGF regulation and they describe, for the first time, a functional link between FOXD1 and implantation/placental diseases. FOXD1 could therefore be used in clinical environments as a molecular biomarker for these diseases in the near future.
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Aborto Habitual/genética , Retardo do Crescimento Fetal/genética , Fatores de Transcrição Forkhead/genética , Predisposição Genética para Doença/genética , Pré-Eclâmpsia/genética , Estudos de Coortes , Feminino , Fatores de Transcrição Forkhead/metabolismo , Humanos , Mutação/genética , Polimorfismo de Nucleotídeo Único/genética , Gravidez , Regiões Promotoras Genéticas/genéticaRESUMO
La litiasis biliar está asociada desarrollo de pancreatitis aguda en el 40% de los casos, sin embargo la sensibilidad diagnóstica de la ecografía abdominal y la tomografía es limitada para hallar esta etiología. En el presente caso se ejemplifica el uso de las enzimas hepáticas aminotransferasas para la predicción del origen biliar en paciente con colecistectomía y ecografía abdominal negativa para colelitiasis y coledocolitiasis. Se presenta un paciente de 55 años, con un cuadro característico de pancreatitis aguda de origen biliar, a quien se hizo el diagnóstico y el tratamiento endoscópico recomendado.
Gallstones are associated with development of acute pancreatitis in 40% of cases, however the diagnostic sensitivities of abdominal ultrasound and CT scans for finding this etiology are limited. The case presented here exemplifies the predictive use of aminotransferase liver enzymes in patients with biliary cholecystectomy but abdominal ultrasound that is negative for cholelithiasis and choledocholithiasis. The case is a 55 year old patient whose clinical picture was consistent with acute biliary pancreatitis. The diagnosis was made and endoscopic treatment was recommended.