RESUMO
Thalidomide is widely used for several diseases; however, it causes malformations in embryos exposed during pregnancy. The complete understanding of the mechanisms by which thalidomide affects the embryo development has not yet been obtained. The phenotypic similarity makes TE a phenocopy of syndromes caused by mutations in ESCO2, SALL4 and TBX5 genes. Recently, SALL4 and TBX5 were demonstrated to be thalidomide targets. To understand if these genes act in the TE development, we sequenced them in 27 individuals with TE; we verified how thalidomide affect them in human pluripotent stem cells (hPSCs) through a differential gene expression (DGE) analysis from GSE63935; and we evaluated how these genes are functionally related through an interaction network analysis. We identified 8 variants in ESCO2, 15 in SALL4 and 15 in TBX5. We compared allelic frequencies with data from ExAC, 1000 Genomes and ABraOM databases; eight variants were significantly different (p < 0.05). Eleven variants in SALL4 and TBX5 were previously associated with cardiac diseases or malformations; however, in TE sample there was no association. Variant effect prediction tools showed 97% of the variants with potential to influence in these genes regulation. DGE analysis showed a significant reduction of ESCO2 in hPSCs after thalidomide exposure.
Assuntos
Acetiltransferases/genética , Proteínas Cromossômicas não Histona/genética , Predisposição Genética para Doença , Proteínas com Domínio T/genética , Teratogênese/genética , Talidomida/efeitos adversos , Fatores de Transcrição/genética , Anormalidades Múltiplas/induzido quimicamente , Anormalidades Múltiplas/genética , Brasil , Linhagem Celular , Anormalidades Craniofaciais/induzido quimicamente , Anormalidades Craniofaciais/genética , Conjuntos de Dados como Assunto , Síndrome da Retração Ocular/induzido quimicamente , Síndrome da Retração Ocular/genética , Ectromelia/induzido quimicamente , Ectromelia/genética , Feminino , Perfilação da Expressão Gênica , Frequência do Gene , Cardiopatias Congênitas/induzido quimicamente , Cardiopatias Congênitas/genética , Comunicação Interatrial/induzido quimicamente , Comunicação Interatrial/genética , Humanos , Hipertelorismo/induzido quimicamente , Hipertelorismo/genética , Hanseníase/tratamento farmacológico , Deformidades Congênitas das Extremidades Inferiores/induzido quimicamente , Deformidades Congênitas das Extremidades Inferiores/genética , Masculino , Mutação , Células-Tronco Pluripotentes , Polimorfismo de Nucleotídeo Único , Gravidez , Complicações na Gravidez/tratamento farmacológico , Mapas de Interação de Proteínas/genética , Teratogênese/efeitos dos fármacos , Deformidades Congênitas das Extremidades Superiores/induzido quimicamente , Deformidades Congênitas das Extremidades Superiores/genéticaRESUMO
Although the thalidomide tragedy occurred more than 50 years ago, the medication is still being used worldwide for different reasons, and several aspects regarding its teratogenicity remain unsolved. Despite the strict regulation implemented, new cases of thalidomide embryopathy (TE) are still being registered in Brazil. Furthermore, the molecular processes that lead to malformations when the embryo is exposed to thalidomide have not yet been fully identified. In this article, we perform a critical analysis of thalidomide's history in Brazil, highlighting aspects of the occurrence of TE over the decades. Finally, we present the main perspectives and challenges for ongoing surveillance and prevention of TE in Brazil. The effective control of dispensing thalidomide, especially in areas where leprosy is endemic, is one of the most important and challenging points. Furthermore, the emergence of thalidomide analogues is fast approaching, and their availability would pose additional concerns. The understanding of the molecular mechanisms and targets of thalidomide in both experimental and human models is essential for generating new insights into teratogenic mechanisms, so that safer thalidomide analogues can be developed.
Assuntos
Anormalidades Múltiplas/fisiopatologia , Doenças Fetais/fisiopatologia , Hanseníase/fisiopatologia , Talidomida/efeitos adversos , Anormalidades Múltiplas/induzido quimicamente , Anormalidades Múltiplas/epidemiologia , Brasil , Anormalidades Congênitas/epidemiologia , Anormalidades Congênitas/fisiopatologia , Doenças Fetais/induzido quimicamente , Doenças Fetais/epidemiologia , Humanos , Hanseníase/complicações , Hanseníase/tratamento farmacológico , Teratogênese/efeitos dos fármacos , Teratogênicos/toxicidadeRESUMO
INTRODUCTION: Thalidomide causes congenital defects in children, such as limb reduction defects. Currently, it is used for a few indications; in Brazil, where leprosy is endemic, thalidomide is used for the treatment of erythema nodosum leprosum, and recent cases of thalidomide embryopathy have been reported. METHODS: We analyzed the frequency of births with phenotypes consistent with thalidomide embryopathy (TEP) and correlated this with the distribution of thalidomide and the prevalence of leprosy between 2005 and 2010 in Brazil. RESULTS: A total of 5,889,210 thalidomide tablets were distributed; the prevalence of limb reduction defects was 1.60 (CI95%: 1.54-1.66) and TEP was 0.11 (CI95%: 0.10-0.13) per 10,000 births. Poisson regression showed an increase in cases of TEP and limb reduction defects per 100,000 tablets dispensed. Clusters and geographical isolates were identified in several regions. CONCLUSIONS: There is a correlation between thalidomide and TEP showing that thalidomide embryopathy should be monitored in countries where this medication is available.
Assuntos
Anormalidades Induzidas por Medicamentos/epidemiologia , Talidomida/efeitos adversos , Brasil/epidemiologia , Prescrições de Medicamentos/estatística & dados numéricos , Feminino , Humanos , Recém-Nascido , Hanseníase/tratamento farmacológico , Hanseníase/epidemiologia , Fenótipo , Gravidez , PrevalênciaRESUMO
Thalidomide is one of the most potent teratogens known to humans. It is currently used for many clinical situations such as treatment of leprosy reactions and multiple myeloma. However, the teratogenic mechanisms by which it produces morphological defects still remain unclear. One of the hypotheses is the blockage of angiogenesis by reduction of nitric oxide (NO). In this study, we evaluated two functional polymorphisms of the endothelial nitric oxide synthase (eNOS) gene which is a constitutively expressed enzyme responsible for production of NO. The promoter -786T>C exon 7 (896G>T) polymorphisms were genotyped using real-time PCR for 28 individuals with thalidomide embryopathy (TE), 27 first-degree relatives of these individuals, and 68 individuals from the general population. Their allele, genotypic, and haplotypic frequencies were compared. A significant difference was observed in the -786T>C polymorphism genotypes (p=0.03) between the groups affected by TE and those unaffected (non-relatives). The TT genotype of the 896G>T polymorphism was observed in 10.7% of those affected and 2.9% of those unaffected, but the difference was not statistically significant (p=0.09). The haplotypic analysis indicated that the wild haplotype -786T/896G was distributed differently in the affected and unaffected groups (p=0.004). These results indicate that the individuals with TE have a higher frequency of alleles associated with lower expression of eNOS, indicating that this may be a genotype susceptible to TE.
Assuntos
Doenças Fetais/induzido quimicamente , Doenças Fetais/genética , Óxido Nítrico Sintase Tipo III/genética , Polimorfismo de Nucleotídeo Único/genética , Talidomida/efeitos adversos , Frequência do Gene , HumanosRESUMO
The thalidomide tragedy of the 1960s resulted in thousands of children being born with severe limb reduction defects (LRD), among other malformations. In Brazil, there are still babies born with thalidomide embryopathy (TE) because of leprosy prevalence, availability of thalidomide, and deficiencies in the control of drug dispensation. Our objective was to implement a system of proactive surveillance to identify birth defects compatible with TE. Along one year, newborns with LRD were assessed in the Brazilian hospitals participating in the Latin-American Collaborative Study of Congenital Malformations (ECLAMC). A phenotype of LRD called thalidomide embryopathy phenotype (TEP) was established for surveillance. Children with TEP born between the years 2000-2008 were monitored, and during the 2007-2008 period we clinically investigated in greater detail all cases with TEP (proactive period). The period from 1982 to 1999 was defined as the baseline period for the cumulative sum statistics. The frequency of TEP during the surveillance period, at 3.10/10,000 births (CI 95%: 2.50-3.70), was significantly higher than that observed in the baseline period (1.92/10,000 births; CI 95%: 1.60-2.20), and not uniformly distributed across different Brazilian regions. During the proactive surveillance (2007-2008), two cases of suspected TE were identified, although the two mothers had denied the use of the drug during pregnancy. Our results suggest that TEP has probably increased in recent years, which coincides with the period of greater thalidomide availability. Our proactive surveillance identified two newborns with suspected TE, proving to be a sensitive tool to detect TE. The high frequency of leprosy and the large use of thalidomide reinforce the need for a continuous monitoring of TEP across Brazil.