Sickle Cell Disease – Current Treatment and New Therapeutical Approaches

Sickle cell disease (SCD) is one of the most common genetic disorders worldwide. It is caused by a point mutation that changes glutamic acid (Glu6) to valine (Val6) in the β chain of hemoglobin. Vaso-occlusion is the most well-known problem associated with SCD. Despite recent advances in understanding the disease at the molecular level, few therapeutic strategies are available. Hydroxyurea is the only drug currently approved by the U.S. Food and Drug Administration for the disease, and it has serious adverse effects and lack of efficacy in some patients. However, new therapeutic approaches are under investigation in the hope of discovering new drugs to treat SCD. These include agents that: a) increase nitric oxide bioavailability; b) modify the rheological properties of the blood; c) bind covalently to hemoglobin; d) prevent hemoglobin dehydration; e) reduce iron overload; and f) induce the expression of gamma globin and fetal hemoglobin. In this chapter, we discuss the current treatment of SCD and the advances made in medicinal chemistry to find new drugs to treat this neglected hematological disease

Uso de compostos derivados ftalimídicos e/ou sulfonamídicos no tratamento de doenças em que há a necessidade de diminuição dos níveis do fator TNF-α e a necessidade de uma fonte exógena de óxido nítrico, compostos derivados ftalimídicos, compostos derivados sulfonamídicos, método de obtenção de um composto derivado sulfonamídico

Em 15/12/2016: Anuidade de pedido de patente de invenção no prazo ordinário.DepositadaA presente invenção se refere ao uso de compostos derivados ftalimídicos e/ou sulfonamídicos com propriedades doadoras de óxido nítrico, os quais apresentam importantes atividades no aumento da expressão gênica de gama globina e atividades anti-inflamatórias e analgésicas, potenciais ao tratamento de doenças hematológicas em que há a necessidade de diminuição dos níveis do fator TNF-α e a necessidade de uma fonte exógena de óxido nítrico. Mais particularmente, a presente invenção descreve o uso de tais derivados ftalimídicos e/ou sulfonamídicos para o tratamento de anemia falciforme. A invenção também apresenta como característica inovadora, a descrição de novos derivados ftalimídicos funcionalizados, desenhados a partir dos protótipos talidomida e hidroxiuréia e planejados racionalmente através da estratégia de hibridação molecular para o tratamento das doenças citadas. A invenção apresenta ainda um novo método de obtenção de um derivado sulfonamídico específico que pode ser utilizado na preparação de um medicamento para tratamento de doenças em que há a necessidade de diminuição dos níveis do fator TNF-α e a necessidade de uma fonte exógena de óxido nítrico

Association Of Nitric Oxide Synthase And Matrix Metalloprotease Single Nucleotide Polymorphisms With Preeclampsia And Its Complications.

Preeclampsia is one of the leading causes of maternal and neonatal morbidity and mortality in the world, but its appearance is still unpredictable and its pathophysiology has not been entirely elucidated. Genetic studies have associated single nucleotide polymorphisms in genes encoding nitric oxide synthase and matrix metalloproteases with preeclampsia, but the results are largely inconclusive across different populations. To investigate the association of single nucleotide polymorphisms (SNPs) in NOS3 (G894T, T-786C, and a variable number of tandem repetitions VNTR in intron 4), MMP2 (C-1306T), and MMP9 (C-1562T) genes with preeclampsia in patients from Southeastern Brazil. This prospective case-control study enrolled 77 women with preeclampsia and 266 control pregnant women. Clinical data were collected to assess risk factors and the presence of severe complications, such as eclampsia and HELLP (hemolysis, elevated liver enzymes, and low platelets) syndrome. We found a significant association between the single nucleotide polymorphism NOS3 T-786C and preeclampsia, independently from age, height, weight, or the other SNPs studied, and no association was found with the other polymorphisms. Age and history of preeclampsia were also identified as risk factors. The presence of at least one polymorphic allele for NOS3 T-786C was also associated with the occurrence of eclampsia or HELLP syndrome among preeclamptic women. Our data support that the NOS3 T-786C SNP is associated with preeclampsia and the severity of its complications.10e013669

Analysis of cyclic necleotides and their associations with the production of fetal hemoglobin in the in vitro erythropoesis of CD34+ cells from patients with sickles cell disease

Orientador: Fernando Ferreira CostaTese (doutorado) - Universidade Estadual de Campinas, Universidade Estadual de Campinas, Faculdade de Ciencias MedicasResumo: A anemia falciforme (AF) é causada pela substituição de uma única base no códon 6 do gene da globina beta (GAG - GTG), levando à produção de hemoglobina S. Estudos recentes têm mostrado que a ativação de guanilato ciclase (GCs) em linhagens de células eritrocíticas e em cultura primária de precursores da série vermelha, causa um aumento na transcrição do gene gama globina e conseqüentemente aumento na produção de hemoglobina fetal (Hb F) (Ikuta et al., 2001). Os nucleotídeos cíclicos, AMPc e GMPc são segundos mensageiros responsáveis por vários sinais extracelulares, incluindo hormônios e neurotransmissores e provavelmente têm papel importante na anemia falciforme regulando a produção de HbF. Este trabalho teve como objetivo, primeiramente, estabelecer um sistema de cultura de células CD 34+ e posteriormente investigar a existência de uma via NO-GMPc/AMPc envolvida na produção de Hb F nos eritrócitos de pacientes com anemia falciforme e verificar se haveria aumento da expressão de GCs e de gama globina nestes mesmos pacientes. Amostras de sangue foram coletadas de voluntários sadios e pacientes com anemia falciforme. Foi utilizado cultura de células CD34+ hematopoéticas de sangue periférico, e ao longo da diferenciação eritróide (7°, 10° e 13° dia) foram coletadas células para citometria de fluxo, citospin, dosagem de GMPc e AMPc e avaliação da expressão gênica de gama globina, GCs alfa e beta subunidades, eNOS, PDE3B, CD36, CD29B e CD49. Observamos que gama globina encontra-se significativamente mais expressa nas culturas falciformes no 13° dia de diferenciação eritróide em relação ao mesmo ponto das culturas controle (2,945 ± 0,303 x 1,445 ± 0,165, respectivamente ,p=0,0095, n=6). A expressão de GCs alfa e beta subunidades, eNOS, PDE3B, CD36, CD29B e CD49 não apresentou diferenças significativas durante a diferenciação eritróide nas culturas falciformes e controles. Os níveis de GMPc nas culturas de pacientes com anemia falciforme estavam significativamente diminuídos ao longo da eritropoese in vitro em comparação com as culturas controles (dia 7: 0,2765 ± 0,0527 x 0,639 ± 0,1221 (p=0,0260); dia 10: 0,1166 ± 0,0219 x 0,268 ± 0,1181; dia 13: 0,037 ± 0,0061 x 0,093 ± 0,0236 (p=0,0152), respectivamente), enquanto o AMPc, encontrava-se elevado no dia 7 das culturas falciformes em relação às culturas controles (dia 7: 72,017 ± 14,849 x 24,987 ± 13,497 (p=0,048); dia 10: 36,3384 ± 16,708 x 8,663 ± 4,591; dia 13: 1,7138 ± 1,093 x 0,680 ± 0,370, respectivamente). O tratamento com BAY 41-2272 (ativador de GCs) em única cultura controle aumentou aproximadamente 3x os níveis de gama globina após 72 h de incubação. Nossos resultados sugerem que as células tronco hematopoéticas dos pacientes com AF são diferentes às dos controles, uma vez que ambas, sob as mesmas condições in vitro expressaram níveis significativamente diferentes de gama globina. Um cross talk entre o GMPc e o AMPc poderia regular a expressão de gama globina ao longo da diferenciação eritróide. Estes dados resultam no primeiro trabalho a demonstrar a expressão basal de gama globina ao longo da diferenciação eritróide de pacientes com anemia falciforme. Desta maneira, o esclarecimento desta via pode contribuir com entendimento da fisiopatologia da anemia falciforme e identificar novos alvos para a regulação farmacológica de hemoglobina fetalAbstract: Sickle cell disease (SCD) is caused by the substitution of a single base in codon 6 of the beta globin gene (GAG-GTG), causing the production of S hemoglobin. Recent studies have demonstrated that the activation of soluble guanylate cyclase (sGC) in erythrocytic cell lines and in primary erythroid precursor culture causes an increase in the transcription of the gamma globin gene and, consequently, an augmented production of fetal hemoglobin (HbF) (Ikuta et al., 2001). The cyclic nucleotides, cAMP and cGMP, are second messengers that are responsible for mediating the signaling of various extracellular signals, including those of hormones and neurotransmitters and, probably, have an important role in SCD by regulating the production of HbF. This study aimed to investigate the existence of a NO-cAMP/cAMP pathway involved in the production of HbF in the erythroid cells of patients with SCD and to determine whether there is an increase in the expression of sGC and of gamma globin in these same patients. Blood samples were collected from healthy volunteers and SCD patients. Peripheral blood hematopoietic CD34+ cell culture was used for the investigation; cells were differentiated into erythroid lineage and cells collected (7th, 10th and 13th day) for flow cytometry, citospin, measurement of cGMP/cAMP and evaluation of the gene expression of gamma globin, the sGC alpha and beta subunits, eNOS, PDE3B, CD36, CD29B and CD49. We demonstrate that gamma globin was significantly more expressed in the cultures of SCD cells on the 13th day of erythroid differentiation compared to the same time point in control cells (2.95 ± 0.30 vs. 1.45 ± 0.17, respectively, p=0.0095, n=6). The expression of sGC alpha and beta subunits, eNOS, PDE3B, CD36, CD29B and CD49 did not present significant differences during erythroid differentiation in control or SCD cell cultures. Levels of cGMP were significantly decreased during erythropoiesis in the SCD cultures compared to control cultures (day 7: 0.277 ± 0.053 vs. 0.639 ± 0.122 (p=0.026); day 10: 0.117 ± 0.022 vs. 0.268 ± 0,118; day 13: 0.037 ± 0.006 vs. 0.093 ± 0.024 (p=0.015), respectively). In contrast, levels of cAMP were significantly increased in the SCD cultures compared to the control cultures (day 7: 72.02 ± 14.85 vs. 24.99 ± 13.50 (p=0.048); day 10: 36.34 ± 16.71 vs. 8.66 ± 4.59; day 13: 1.74 ± 1.09 vs. 0.680 ± 0.370, respectively). Treatment with BAY 41-2272 (sGC inducer) in a single control cell culture increased the levels of gamma globin by approximately 3 times after 72h of incubation. Results suggest that the hematopoietic stem cells of SCD patients are significantly and constitutively different to those of controls, since under the same in vitro culture conditions, differentiation of the two cell types results in consistently significant different levels of gamma globin. A cross talk between cGMP and cAMP may regulate the expression of gamma globin during erythroid differentiation in patients with SCD. Thus, further investigation of this pathway may contribute to the understanding of the physiopathology of the disease and identify new targets for the pharmacological regulation of HbF productionDoutoradoMedicina ExperimentalDoutor em Fisiopatologia Medic

Indicadores Bioquímicos do Infarto Agudo do Miocárdio/Biochemical Indicators of Acute Myocardial Infarction

Este artigo é uma revisão sobre os indicadores bioquímicos mais utilizados para diagnosticar Infarto Agudo do Miocárdio. Aborda-se rapidamente as doenças cardiovasculares, dando ênfase ao Infarto Agudo do Miocárdio, seus fatores de risco, prevenção e tratamento. Apresentam-se os marcadores cardíacos mais indicados, qualificando-os conforme suas especificidade e sensibilidade para o diagnóstico, concluindo que é importante a dosagem dos diferentes marcadores no diagnóstico do infarto agudo do miocárdio, em diferentes horários, pois estes se elevam conforme sua sensibilidade e especificidade. No entanto, a sintomatologia clínica e outros exames, como eletrocardiogramas devem estar presentes, possibilitando um diagnóstico mais seguro e um melhor monitoramento do paciente infartado.  Palavras Chave: marcadores cardíacos, indicadores bioquímicos, infarto agudo do miocárdio.  This article is an overview of the most widely used biochemical markers to diagnose acute myocardial infarction. Covers quickly cardiovascular diseases, with emphasis on acute myocardial infarction, its risk factors, prevention and treatment. Presents the most appropriate cardiac markers, qualifying them according to their specificity and sensitivity for the diagnosis, concluding that it is important to the dosage of different markers in the diagnosis of acute myocardial infarction at different times, because they rise as their sensitivity and specificity. However, the clinical symptoms and other tests such as electrocardiograms must be present, providing a more secure diagnosis and better patient monitoring myocardium.  Key words: cardiac markers, biochemical markers, acute myocardial infarction

Clinical relevance of heterozygosis for aceruloplasminemia

Aceruloplasminemia is a rare form of brain iron overload of autosomal recessive inheritance that results from mutations in the CP gene, encoding the iron oxidase ceruloplasmin. Homozygous aceruloplasminemia causes progressive neurodegenerative disease, anemia, and diabetes, and is usually diagnosed late in life upon investigation of anemia, high ferritin, or movement disorders, but its heterozygous state is less characterized and believed to be silent. Here we report two heterozygotes for new mutations causing aceruloplasminemia from whom peripheral blood samples were collected for complete blood counts, iron studies, and genotyping by automated sequencing. We then performed a systematic review of preview reports of heterozygotes with data on genotype and clinical findings. Heterozygosity for aceruloplasminemia invariably causes reduced ceruloplasmin levels, and similarly to previews reports in the literature, our cases did not present with anemia. Mild hyperferritinemia was found only in two reports. Nevertheless, 5 out of 11 variants have been associated with significant neurological symptoms despite the presence of one wild-type alelle. This review contributes to better genetic counseling of heterozygotes for CP gene variants and supports that measuring ceruloplasmin levels may be useful when investigating patients with movement disorders or rare cases of unexplained high ferritin1804266271FAPESP – Fundação de Amparo à Pesquisa Do Estado De São Paulo2014/00984-3; 2016/08072-