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1.
Cells ; 8(4)2019 04 19.
Artigo em Inglês | MEDLINE | ID: mdl-31010158

RESUMO

Parkinson's disease (PD) is the second most common degenerative disorder. Although the disease was described more than 200 years ago, its pathogenetic mechanisms have not yet been fully described. In recent years, the discovery of the association between mutations of the GBA gene (encoding for the lysosomal enzyme glucocerebrosidase) and PD facilitated a better understating of this disorder. GBA mutations are the most common genetic risk factor of the disease. However, mutations of this gene can be found in different phenotypes, such as Gaucher's disease (GD), PD, dementia with Lewy bodies (DLB) and rapid eye movements (REM) sleep behavior disorders (RBDs). Understanding the pathogenic role of this mutation and its different manifestations is crucial for geneticists and scientists to guide their research and to select proper cohorts of patients. Moreover, knowing the implications of the GBA mutation in the context of PD and the other associated phenotypes is also important for clinicians to properly counsel their patients and to implement their care. With the present review we aim to describe the genetic, clinical, and therapeutic features related to the mutation of the GBA gene.


Assuntos
Doença de Gaucher/genética , Glucosilceramidase/genética , Doença de Parkinson/genética , Doença de Gaucher/terapia , Humanos , Doença por Corpos de Lewy/genética , Mutação , Doença de Parkinson/terapia , Fenótipo , Fatores de Risco
2.
J Pediatr Endocrinol Metab ; 32(5): 533-536, 2019 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-31026225

RESUMO

Background Gaucher disease (GD) is a lysosomal storage disorder caused by autosomal recessive mutations in the glucocerebrosidase (GBA) gene, which encodes acid ß-glucosidase. GD type 3c is a rare group characterised by cardiovascular involvement, and homozygous D448H is the most frequent mutation. Case presentation We describe two patients who had homozygous D448H mutations. The index patient had hepatosplenomegaly, liver insufficiency and cardiac involvement and her sister had severe cardiac involvement with cardiomyopathy and diffuse aortic calcification. The index case's liver was transplanted at the age of 6 months from a related donor and her sister who had severe cardiovascular disease died at the age of 12 years. Conclusions Our patients had clinical variability. We need to discuss whether liver involvement could be the initial signs in patients with GD type 3c.


Assuntos
Doenças Cardiovasculares/patologia , Doença de Gaucher/complicações , Glucosilceramidase/genética , Hepatopatias/patologia , Mutação , Doenças Cardiovasculares/etiologia , Criança , Feminino , Doença de Gaucher/enzimologia , Doença de Gaucher/genética , Homozigoto , Humanos , Lactente , Hepatopatias/etiologia , Prognóstico , Irmãos
3.
Neurobiol Dis ; 127: 563-569, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30981829

RESUMO

Bi-allelic mutations in the glucocerebrosidase gene (GBA1) cause Gaucher's disease, the most common human lysosomal storage disease. We previously reported a marked increase in miR-155 transcript levels and early microglial activation in a zebrafish model of Gaucher's disease (gba1-/-). miR-155 is a master regulator of inflammation and has been implicated in a wide range of different neurodegenerative disorders. The observed miR-155 upregulation preceded the subsequent development of widespread pathology with marked neuroinflammation, closely resembling human Gaucher's disease pathology. We now report similar increases of miR-155 expression in mammalian models of GD, confirming that miR-155 upregulation is a shared feature in glucocerebrosidase (GCase) deficiency across different species. Using CRISPR/Cas9 mutagenesis we then generated a miR-155 mutant zebrafish line (miR-155-/-) with completely abolished miR-155 expression. Unexpectedly, loss of miR-155 did not mitigate either the reduced lifespan or the robust inflammatory phenotypes of gba1-/- mutant zebrafish. Our data demonstrate that neither neuroinflammation nor disease progression in GCase deficiency are dependent on miR-155 and suggest that miR-155 inhibition would not be a promising therapeutic target in Gaucher's disease.


Assuntos
Encefalite/metabolismo , Doença de Gaucher/metabolismo , MicroRNAs/metabolismo , Degeneração Neural/metabolismo , Animais , Animais Geneticamente Modificados , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Citocinas/metabolismo , Modelos Animais de Doenças , Progressão da Doença , Encefalite/genética , Encefalite/patologia , Doença de Gaucher/genética , Doença de Gaucher/patologia , Glucosilceramidase/genética , Glucosilceramidase/metabolismo , Camundongos , MicroRNAs/genética , Mutação , Degeneração Neural/genética , Degeneração Neural/patologia , Neurônios/metabolismo , Neurônios/patologia , Regulação para Cima , Peixe-Zebra
4.
J Gastrointestin Liver Dis ; 28(1): 121-123, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30851181

RESUMO

Gaucher's disease and alpha-1 antitrypsin deficiency are genetic diseases that can cause different kinds of liver damage, but are rarely associated with cirrhosis. Here, we describe the case of a patient with both diseases who presented with cirrhosis, followed by liver failure and death. Although the interaction between these two diseases remains unclear, we suspect the involvement of an epigenetic factor in the evolution of the aggressive liver disease.


Assuntos
Epigênese Genética , Doença de Gaucher/genética , Cirrose Hepática/genética , Falência Hepática/genética , Deficiência de alfa 1-Antitripsina/genética , Progressão da Doença , Terapia de Reposição de Enzimas , Evolução Fatal , Feminino , Doença de Gaucher/complicações , Doença de Gaucher/diagnóstico , Predisposição Genética para Doença , Humanos , Cirrose Hepática/diagnóstico , Falência Hepática/diagnóstico , Pessoa de Meia-Idade , Fenótipo , Fatores de Risco , Resultado do Tratamento , Deficiência de alfa 1-Antitripsina/complicações , Deficiência de alfa 1-Antitripsina/diagnóstico
5.
BMC Med Genet ; 20(1): 31, 2019 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-30764785

RESUMO

BACKGROUND: Gaucher disease is a rare pan-ethnic, lysosomal storage disorder resulting due to beta-Glucosidase (GBA1) gene defect. This leads to the glucocerebrosidase enzyme deficiency and an increased accumulation of undegraded glycolipid glucocerebroside inside the cells' lysosomes. To date, nearly 460 mutations have been described in the GBA1 gene. With the aim to determine mutations spectrum and molecular pathology of Gaucher disease in India, the present study investigated one hundred unrelated patients (age range: 1 day to 31 years) having splenomegaly, with or without hepatomegaly, cytopenia and bone abnormality in some of the patients. METHODS: The biochemical investigation for the plasma chitotriosidase enzyme activity and ß-Glucosidase enzyme activity confirmed the Gaucher disease. The mutations were identified by screening the patients' whole GBA gene coding region using bidirectional Sanger sequencing. RESULTS: The biochemical analysis revealed a significant reduction in the ß-Glucosidase activity in all patients. Sanger sequencing established 71 patients with homozygous mutation and 22 patients with compound heterozygous mutation in GBA1 gene. Lack of identification of mutations in three patients suggests the possibility of either large deletion/duplication or deep intronic variations in the GBA1 gene. In four cases, where the proband died due to confirmed Gaucher disease, the parents were found to be a carrier. Overall, the study identified 33 mutations in 100 patients that also covers four missense mutations (p.Ser136Leu, p.Leu279Val, p.Gly383Asp, p.Gly399Arg) not previously reported in Gaucher disease patients. The mutation p.Leu483Pro was identified as the most commonly occurring Gaucher disease mutation in the study (62% patients). The second common mutations identified were p.Arg535Cys (7% patients) and RecNcil (7% patients). Another complex mutation Complex C was identified in a compound heterozygous status (3% patients). The homology modeling of the novel mutations suggested the destabilization of the GBA protein structure due to conformational changes. CONCLUSIONS: The study reports four novel and 29 known mutations identified in the GBA1 gene in one-hundred Gaucher patients. The given study establishes p.Leu483Pro as the most prevalent mutation in the Indian patients with type 1 Gaucher disease that provide new insight into the molecular basis of Gaucher Disease in India.


Assuntos
Grupo com Ancestrais do Continente Europeu/genética , Doença de Gaucher/genética , Glucosilceramidase/genética , Mutação , Análise de Sequência de DNA/métodos , Adolescente , Adulto , Substituição de Aminoácidos , Criança , Pré-Escolar , Éxons , Feminino , Doença de Gaucher/metabolismo , Glucosilceramidase/química , Glucosilceramidase/metabolismo , Humanos , Índia , Lactente , Recém-Nascido , Masculino , Modelos Moleculares , Homologia Estrutural de Proteína , Adulto Jovem
6.
Adv Protein Chem Struct Biol ; 114: 315-339, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30635084

RESUMO

Gaucher's disease (GD) is the most commonly known lysosomal disorder that occurs due to mutations in the ß-glucocerebrosidase (GBA) protein. Our previous findings (Thirumal Kumar, Eldous, Mahgoub, George Priya Doss, Zayed, 2018) and other reports concluded that the mutations N370S and L444P are the most significant mutations that could cause disruptions in protein stability and structure. These disruptions lead to protein misfolding and result in a diseased condition. Enzyme Replacement Therapy (ERT) and Pharmacological chaperone therapy (PCT) are currently used to treat GD caused by mutations in the GBA protein. The extreme disparity in cost between ERT and chaperone therapy, shifted the attention toward chaperone therapy. The most common chaperones in the market and trial phases to treat GD are Isofagomine, Miglustat, Eliglustat, NN-DNJ, and Ambroxol. In the era of personalized medicine, it is often necessary to understand the drug likeliness of each chaperone. In this context, the present study utilized molecular docking analysis to understand the interaction behavior of the chaperone toward the native and the two mutants N370S and L444P. The molecular dynamics simulation analyses performed on chaperones (NN-DNJ and Ambroxol) interaction showed that the chaperone NN-DNJ possesses better affinity toward the protein with N370S mutation whereas chaperone Ambroxol showed better activity against both the significant mutations (N370S and L444P). This study is expected to serve as a platform for drug repurposing.


Assuntos
1-Desoxinojirimicina/análogos & derivados , Ambroxol/farmacologia , Doença de Gaucher/tratamento farmacológico , Doença de Gaucher/genética , Glucosilceramidase/antagonistas & inibidores , Chaperonas Moleculares/farmacologia , Mutação , 1-Desoxinojirimicina/química , 1-Desoxinojirimicina/farmacologia , Ambroxol/química , Glucosilceramidase/genética , Glucosilceramidase/metabolismo , Humanos , Modelos Moleculares , Chaperonas Moleculares/química
7.
Clin Chim Acta ; 492: 1-6, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30695688

RESUMO

BACKGROUND: Chitotriosidase (ChT) is used as a biomarker for the follow-up of patients with Gaucher disease (GD), once his activity is extremely elevated and declines during ERT. However, some variants in the CHIT1 gene affect ChT activity. METHODS: To assess association between ChT genotype, and clinical/biochemical features of GD were performed CHIT1 genotyping for: c.1049_1072dup24, p.Gly102Ser, p.Gly354Arg, c.1155_1156 + 2delGAGT, c.1156 + 5_1156 + 8delGTAA, p.Ala442Val/Gly and the rearrangement delE/I-10. RESULTS: Were evaluated 42 patients with GD from Southern Brazil. Pretreatment ChT activity was available for 32 patients. Allelic frequencies found for dup24, p.Gly102Ser and p.Ala442Gly were 0.14, 0.32 and 0.12, respectively. Only one patient presented reduced ChT activity (dup24 homozygous). Comparison between wild homozygous and heterozygous for dup24 showed that both differ in relation to the ChT activity before (15,230 vs 6936 nmol/h/mL, p < .001), but not after treatment (5212 vs 3045 nmol/h/mL, p = .227). CONCLUSIONS: Pretreatment ChT activity was not correlated with clinical/biochemical features. There was a reduction of 63% in the ChT activity after 12 months on treatment (p < .001). There is no evidence that higher ChT levels are associated with a more severe symptomatology in untreated GD patients. The pretreatment ChT levels appear to be mainly dependent on the presence/absence of the dup24 allele.


Assuntos
Doença de Gaucher/enzimologia , Doença de Gaucher/genética , Genótipo , Hexosaminidases/genética , Fenótipo , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Feminino , Frequência do Gene , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem
8.
Mol Genet Genomic Med ; 7(3): e564, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30637984

RESUMO

BACKGROUND: Mutations in GBA cause Gaucher disease when biallelic and are strong risk factors for Parkinson's disease when heterozygous. GBA analysis is complicated by the nearby pseudogene. We aimed to design and validate a method for sequencing GBA using long reads. METHODS: We sequenced GBA on the Oxford Nanopore MinION as an 8.9 kb amplicon from 102 individuals, including patients with Parkinson's and Gaucher diseases. We used NanoOK for quality metrics, NGMLR to align data (after comparing with GraphMap), Nanopolish and Sniffles to call variants, and WhatsHap for phasing. RESULTS: We detected all known missense mutations in these samples, including the common p.N409S (N370S) and p.L483P (L444P) in multiple samples, and nine rarer ones, as well as a splicing and a truncating mutation, and intronic SNPs. We demonstrated the ability to phase mutations, confirm compound heterozygosity, and assign haplotypes. We also detected two known risk variants in some Parkinson's patients. Rare false positives were easily identified and filtered, with the Nanopolish quality score adjusted for the number of reads a very robust discriminator. In two individuals carrying a recombinant allele, we were able to detect and fully define it in one carrier, where it included a 55-base pair deletion, but not in another one, suggesting a limitation of the PCR enrichment method. Missense mutations were detected at the correct zygosity, except for the case where the RecNciI one was missed. CONCLUSION: The Oxford Nanopore MinION can detect missense mutations and an exonic deletion in this difficult gene, with the added advantages of phasing and intronic analysis. It can be used as an efficient research tool, but additional work is required to exclude all recombinants.


Assuntos
Doença de Gaucher/genética , Testes Genéticos/métodos , Glucosilceramidase/genética , Mutação de Sentido Incorreto , Análise de Sequência de DNA/métodos , Doença de Gaucher/diagnóstico , Testes Genéticos/normas , Humanos , Kit de Reagentes para Diagnóstico/normas , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Análise de Sequência de DNA/normas
9.
Stem Cell Res ; 35: 101336, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30606667

RESUMO

Gaucher disease is the most common autosomal recessive lysosomal storage disorder, caused by mutations in the ß-glucocerebrosidase gene GBA. Here we describe generation of iPSC from skin-derived fibroblasts from two unrelated individuals with neuronopathic forms of Gaucher disease. The donor for line iPSC-GBA-1, a 21 month old girl, carried the recurring GBA mutation c.1448 T > C, p.Leu483Pro at homozygous state; fibroblasts for line iPS-GBA-2 were obtained from a 4 year old girl compound heterozygous for the GBA mutations c.667 T > C, p.Trp223Arg and c.1226A > G, p.Asn409Ser. iPSCs were developed using integration free episomal vectors (OCT4, KLF4; L-MYC, SOX2 (OSKM) and LIN28). Resource table.


Assuntos
Linhagem Celular , Doença de Gaucher/metabolismo , Células-Tronco Pluripotentes Induzidas/metabolismo , Alelos , Pré-Escolar , Feminino , Doença de Gaucher/genética , Doença de Gaucher/patologia , Glucosilceramidase/metabolismo , Homozigoto , Humanos , Células-Tronco Pluripotentes Induzidas/patologia , Lactente , Mutação
10.
Biochem J ; 476(2): 261-274, 2019 01 25.
Artigo em Inglês | MEDLINE | ID: mdl-30578288

RESUMO

Gaucher disease (GD) is a rare lysosomal storage disorder caused by mutations in the GBA1 gene, encoding the lysosome-resident glucocerebrosidase enzyme involved in the hydrolysis of glucosylceramide. The discovery of an association between mutations in GBA1 and the development of synucleinopathies, including Parkinson disease, has directed attention to glucocerebrosidase as a potential therapeutic target for different synucleinopathies. These findings initiated an exponential growth in research and publications regarding the glucocerebrosidase enzyme. The use of various commercial and custom-made glucocerebrosidase antibodies has been reported, but standardized in-depth validation is still not available for many of these antibodies. This work details the evaluation of several previously reported glucocerebrosidase antibodies for western blot analysis, tested on protein lysates of murine gba+/+ and gba-/- immortalized neurons and primary human wild-type and type 2 GD fibroblasts.


Assuntos
Anticorpos/química , Western Blotting , Fibroblastos/enzimologia , Doença de Gaucher/enzimologia , Glucosilceramidase/metabolismo , Doença de Parkinson/enzimologia , Animais , Linhagem Celular Transformada , Fibroblastos/patologia , Doença de Gaucher/genética , Doença de Gaucher/patologia , Glucosilceramidase/genética , Humanos , Camundongos , Camundongos Knockout , Doença de Parkinson/genética , Doença de Parkinson/patologia
11.
Int J Hematol ; 109(3): 361-365, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30456712

RESUMO

Gaucher disease (GD) is caused by a hereditary deficiency of glucocerebrosidase, resulting in accumulation of glucosylceramide and potentially manifesting as hepatosplenomegaly. We report the case of a 15-month-old boy with chronic neuronopathic GD. The patient had prolonged anemia despite continued iron supplementation for 3 months. White blood count (WBC), hemoglobin (Hb), platelet count, and corrected reticulocyte count were 3,300 /µL, 8.7 g/dL, 90,000 /µL, and 0.55, respectively. The patient had microcytic hypochromic anemia with mildly elevated ferritin. Physical examination revealed hepatosplenomegaly. Bone-marrow aspiration showed sheets of Gaucher cells. Glucocerebrosidase activity in monocytes was significantly lower than normal. Genetic analysis revealed a homozygous L444P mutation of GBA, and he was diagnosed with type 1 GD. Enzyme replacement treatment (ERT) consisting of imiglucerase was initiated and was effective; WBC, Hb, and platelet count gradually normalized and the hepatosplenomegaly improved. However, when the patient entered elementary school, he showed mild impaired cognitive function, and supranuclear gaze palsy occurred the same year. He was ultimately diagnosed with type 3 GD and continued ERT. Pediatric hemato-oncologists should be aware of GD, especially when patients exhibit anemia refractory to iron therapy, radiologic bone deformity, neurologic signs or symptoms, and growth retardation.


Assuntos
Anemia Hipocrômica , Terapia de Reposição de Enzimas , Doença de Gaucher , Glucosilceramidase/uso terapêutico , Substituição de Aminoácidos , Anemia Hipocrômica/sangue , Anemia Hipocrômica/diagnóstico , Anemia Hipocrômica/tratamento farmacológico , Anemia Hipocrômica/genética , Contagem de Células Sanguíneas , Medula Óssea/metabolismo , Doença de Gaucher/sangue , Doença de Gaucher/diagnóstico , Doença de Gaucher/tratamento farmacológico , Doença de Gaucher/genética , Glucosilceramidase/genética , Glucosilceramidase/metabolismo , Hemoglobinas/metabolismo , Humanos , Lactente , Masculino , Mutação de Sentido Incorreto
12.
J Neurol ; 266(1): 92-101, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30382391

RESUMO

BACKGROUND: Patients with Gaucher Disease (GD) exhibit three phenotypes, including type 1 (non-neuronopathic), type 2 (acute neuronopathic), and type 3 (subacute neuronopathic). AIM: Identifying which GBA changes represent benign polymorphisms and which may result in disease-causing mutations is essential for diagnosis and genotype/phenotype correlations but is often challenging. RESULTS: Here, we describe a patient with type 3 GD, presenting with drug-resistant epilepsy, who bears a set of GBA polymorphic variants including the novel c.363A > G (Gly82Gly) synonymous mutation. In silico predictions, mRNA and functional studies revealed that the new Gly82Gly mutation causes skipping of GBA exon 4, leading to a severe reduction of the wild type GBA mRNA. This is the first report of a synonymous change causing GD through loss of an exonic splicing enhancer sequence. The synonymous mutation is in trans with the Asn188Ser missense mutation, thus making the Asn188Ser responsible for the patient's phenotype and strengthening the association of Asn188Ser with the particular neurological phenotype of type 3 GD. CONCLUSION: We strengthen the association of Asn188Ser with the type 3 GD phenotype and progressive myoclonus epilepsy. Our data confirm that in silico predictions and mRNA analysis are mandatory in discriminating pathological mutations from the background of harmless polymorphisms, especially synonymous changes.


Assuntos
Epilepsia Resistente a Medicamentos/genética , Doença de Gaucher/genética , Glucosilceramidase/genética , Mutação , Epilepsias Mioclônicas Progressivas/genética , Adolescente , Encéfalo/fisiopatologia , Epilepsia Resistente a Medicamentos/enzimologia , Epilepsia Resistente a Medicamentos/fisiopatologia , Éxons , Feminino , Fibroblastos/enzimologia , Doença de Gaucher/enzimologia , Doença de Gaucher/fisiopatologia , Humanos , Epilepsias Mioclônicas Progressivas/enzimologia , Epilepsias Mioclônicas Progressivas/fisiopatologia , Fenótipo
13.
Eur J Neurol ; 26(3): 388-e29, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30315684

RESUMO

The association between Gaucher disease (GD) and Parkinson disease (PD) has been described for almost two decades. In the biallelic state (homozygous or compound heterozygous) mutations in the glucocerebrosidase gene (GBA) may cause GD, in which glucosylceramide, the sphingolipid substrate of the glucocerebrosidase enzyme (GCase), accumulates in visceral organs leading to a number of clinical phenotypes. In the biallelic or heterozygous state, GBA mutations increase the risk for PD. Mutations of the GBA allele are the most significant genetic risk factor for idiopathic PD, found in 5%-20% of idiopathic PD cases depending on ethnicity. The neurological consequences of GBA mutations are reviewed and the proposition that GBA mutations result in a disparate but connected range of clinically and pathologically related neurological features is discussed. The literature relating to the clinical, biochemical and genetic basis of GBA PD, type 1 GD and neuronopathic GD is considered highlighting commonalities and distinctions between them. The evidence for a unifying disease mechanism is considered.


Assuntos
Doença de Gaucher/genética , Doença de Gaucher/fisiopatologia , Glucosilceramidase/genética , Doença de Parkinson/genética , Doença de Parkinson/fisiopatologia , Humanos
14.
Mov Disord ; 34(1): 9-21, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30589955

RESUMO

Glucocerebrosidase is a lysosomal enzyme. The characterization of a direct link between mutations in the gene coding for glucocerebrosidase (GBA1) with the development of Parkinson's disease and dementia with Lewy bodies has heightened interest in this enzyme. Although the mechanisms through which glucocerebrosidase regulates the homeostasis of α-synuclein remains poorly understood, the identification of reduced glucocerebrosidase activity in the brains of patients with PD and dementia with Lewy bodies has paved the way for the development of novel therapeutic strategies directed at enhancing glucocerebrosidase activity and reducing α-synuclein burden, thereby slowing down or even preventing neuronal death. Here we reviewed the current literature relating to the mechanisms underlying the cross talk between glucocerebrosidase and α-synuclein, the GBA1 mutation-associated clinical phenotypes, and ongoing therapeutic approaches targeting glucocerebrosidase. © 2018 International Parkinson and Movement Disorder Society.


Assuntos
Glucosilceramidase/genética , Lisossomos/genética , Medicina de Precisão , /genética , Doença de Gaucher/genética , Humanos , Lisossomos/metabolismo , Medicina de Precisão/métodos , alfa-Sinucleína/genética , alfa-Sinucleína/metabolismo
16.
EBioMedicine ; 38: 142-153, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30497978

RESUMO

BACKGROUND: Gaucher Disease is caused by mutations of the GBA gene which encodes the lysosomal enzyme acid beta-glucosidase (GCase). GBA mutations commonly affect GCase function by perturbing its protein homeostasis rather than its catalytic activity. Heat shock proteins are well known cytoprotective molecules with functions in protein homeostasis and lysosomal function and their manipulation has been suggested as a potential therapeutic strategy for GD. The investigational drug arimoclomol, which is in phase II/III clinical trials, is a well-characterized HSP amplifier and has been extensively clinically tested. Importantly, arimoclomol efficiently crosses the blood-brain-barrier presenting an opportunity to target the neurological manifestations of GD, which remains without a disease-modifying therapy. METHODS: We used a range of biological and biochemical in vitro assays to assess the effect of arimoclomol on GCase activity in ex vivo systems of primary fibroblasts and neuronal-like cells from GD patients. FINDINGS: We found that arimoclomol induced relevant HSPs such as ER-resident HSP70 (BiP) and enhanced the folding, maturation, activity, and correct cellular localization of mutated GCase across several genotypes including the common L444P and N370S mutations in primary cells from GD patients. These effects where recapitulated in a human neuronal model of GD obtained by differentiation of multipotent adult stem cells. INTERPRETATION: These data demonstrate the potential of HSP-targeting therapies in GCase-deficiencies and strongly support the clinical development of arimoclomol as a potential therapeutic option for the neuronopathic forms of GD. FUNDING: The research was funded by Orphazyme A/S, Copenhagen, Denmark.


Assuntos
Glucosilceramidase/química , Glucosilceramidase/metabolismo , Hidroxilaminas/farmacologia , Lisossomos/metabolismo , Redobramento de Proteína/efeitos dos fármacos , Linhagem Celular , Retículo Endoplasmático/metabolismo , Ativação Enzimática/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Doença de Gaucher/genética , Doença de Gaucher/metabolismo , Glucosilceramidase/genética , Complexo de Golgi/metabolismo , Proteínas de Choque Térmico/metabolismo , Humanos , Mutação , Neurônios , Processamento de Proteína Pós-Traducional , Transporte Proteico
17.
Medicine (Baltimore) ; 97(47): e13161, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30461613

RESUMO

RATIONALE: Gaucher disease (GD), characterized by glucosylceramide accumulation in the macrophage-monocyte system, is caused by glucosidase b acid (GBA) gene mutations which lead to the deficiency of lysosomal enzyme glucocerebrosidase. The mutation spectrum of GBA in Chinese patients is quite different from those seen in Jewish and non-Jewish Caucasian patients. Thus, it is relatively hard to diagnose GD in Chinese. PATIENT CONCERNS: A 24-year-old Chinese female with intermittent abdominal distension and progressive decrease in strength but without neurologic symptoms was initially referred for femoral head necrosis on the right feet. Laboratory examinations results indicated panhematopenia. Bone marrow aspiration smear and biopsy specimen found typical "wrinkled" Gaucher cells. Molecular-genetic testing of GBA gene revealed 3 mutations including R159W (c. 475 C > T), V1230G (c. 689T > G), and G241A (c. 721G > A). DIAGNOSES: On the basis of these findings and clinical manifestations, the final diagnosis of type 1 GD was made. INTERVENTIONS: Enzyme replacement therapy (ERT) with velaglucerase α was carried out after the diagnosis of type 1 GD. OUTCOMES: The platelet and hemoglobin levels were restored by ERT. LESSONS: To our knowledge, this is the first report of GD patient carrying 3 mutations in Chinese. These mutations in GBA in the present case imply a potential pool of patients with GD with this mutation in Chinese.


Assuntos
Grupo com Ancestrais do Continente Asiático/genética , Doença de Gaucher/genética , Glucosilceramidase/genética , Mutação , Terapia de Reposição de Enzimas , Éxons , Feminino , Doença de Gaucher/complicações , Doença de Gaucher/tratamento farmacológico , Glucosilceramidase/uso terapêutico , Humanos , Debilidade Muscular/etiologia , Adulto Jovem
18.
Expert Rev Clin Pharmacol ; 11(12): 1183-1194, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30444430

RESUMO

INTRODUCTION: Gaucher disease (GD) is an autosomal recessive disorder resulting from the deficiency of the lysosomal enzyme glucocerebrosidase (b-glucosidase), associated with varying degrees of visceral, bone and central nervous system pathology, leading to wide phenotypic diversity. Response to therapy and clinical outcomes are very different between the three clinical subtypes - non-neuronopathic, acute neuronopathic, and chronic neuronopathic forms; hence a definitive clinical diagnosis is essential. The availability of two therapeutic options, i.e. enzyme replacement and substrate reduction, has transformed the natural course of the disease. As pre-treatment disease severity clearly impacts results of therapy, early diagnosis and initiation of treatment especially in the pediatric population are keys to achieving an optimal outcome. Areas covered: We reviewed the literature concerning the treatment of GD focusing on pediatric presentations, various pharmacological treatment options and recommendations for management goals. A PubMed literature search was performed for relevant publications between 1991 and September 2018. Expert commentary: The approval of enzyme replacement therapy (ERT) for GD in the pediatric age group has significantly altered the course of the disease, especially for non-neuronopathic and chronic neuronopathic forms, as ERT does not cross the blood-brain barrier. Early diagnosis, regular follow-up and early initiation of treatment can thus prevent some irreversible complications and improve patient quality of life.


Assuntos
Terapia de Reposição de Enzimas/métodos , Doença de Gaucher/tratamento farmacológico , Glucosilceramidase/administração & dosagem , Criança , Doença de Gaucher/genética , Doença de Gaucher/fisiopatologia , Glucosilceramidase/deficiência , Humanos , Qualidade de Vida
19.
BMC Med Genet ; 19(1): 178, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30285649

RESUMO

BACKGROUND: Gaucher disease is a rare pan-ethnic disorder which occurs due to an increased accumulation of undegraded glycolipid glucocerebroside inside the cells' lysosomes. A beta-Glucosidase (GBA) gene defect results in glucocerebrosidase enzyme deficiency. Though the disease is mainly diagnosed in childhood, the adult manifestation is often missed or identified late due to the failure to recognize the heterogeneous clinical presentation. The present study includes seven unrelated Indian adult patients (age range: 20-40 years) having splenomegaly, with or without hepatomegaly, cytopenia and bone abnormality. METHODS: The biochemical investigation implicated measuring plasma chitotriosidase enzyme activity followed by confirmatory test of ß-Glucosidase enzyme activity from the leukocytes. The molecular characterization involved patients' initial screening for the common Gaucher mutation (Leu444Pro). Later, all patients were subjected to whole GBA gene coding region study using bidirectional Sanger sequencing. The population screening for common Gaucher disease mutation (Leu444Pro) was executed in 1200 unrelated and healthy Indian subjects by Restriction Fragment Length Polymorphism-Polymerase Chain Reaction technique. The allele frequency was calculated using Hardy-Weinberg formula. RESULTS: The biochemical analysis revealed a significant reduction in the ß-Glucosidase activity in all the patients. Also, an elevated level of plasma Chitotriosidase activity in five patients supported their diagnosis of Gaucher disease. Sanger sequencing established four patients with homozygous variation and three patients with compound heterozygous variation in GBA gene. This study uncovers two missense variants (Ala448Thr and Val17Gly) not previously reported in Gaucher disease patients. Also the known mutations like Leu444Pro, Arg329Cys, Asp315Asn, Ser125Arg, and Arg395Cys were identified in these patients. The homology modeling suggested the destabilization of the protein structure due to novel variants. The Leu444Pro mutation screening in the Indian population spotted two people as a carrier. This emerged the carrier frequency of 1:600 along with wild-type allele frequency 0.97113 and mutant allele frequency 0.02887. CONCLUSIONS: The study reports novel and known variants identified in the GBA gene in seven adult patients. The given study is the first report on the carrier frequency of the Leu444Pro mutant allele in an Indian population which will help understanding the burden and susceptibility of Gaucher disease to affect next generation in India.


Assuntos
Doença de Gaucher/genética , Hepatomegalia/genética , Mutação , Esplenomegalia/genética , beta-Glucosidase/genética , Adulto , Alelos , Sequência de Aminoácidos , Sequência de Bases , Portador Sadio , Criança , Análise Mutacional de DNA , Éxons , Feminino , Doença de Gaucher/diagnóstico , Doença de Gaucher/enzimologia , Doença de Gaucher/patologia , Expressão Gênica , Frequência do Gene , Glucosilceramidas/metabolismo , Hepatomegalia/diagnóstico , Hepatomegalia/enzimologia , Hepatomegalia/patologia , Hexosaminidases/sangue , Hexosaminidases/genética , Humanos , Índia , Lisossomos/enzimologia , Lisossomos/patologia , Masculino , Estrutura Secundária de Proteína , Índice de Gravidade de Doença , Esplenomegalia/diagnóstico , Esplenomegalia/enzimologia , Esplenomegalia/patologia , beta-Glucosidase/química , beta-Glucosidase/metabolismo
20.
Nat Rev Dis Primers ; 4(1): 27, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30275469

RESUMO

Lysosomal storage diseases (LSDs) are a group of over 70 diseases that are characterized by lysosomal dysfunction, most of which are inherited as autosomal recessive traits. These disorders are individually rare but collectively affect 1 in 5,000 live births. LSDs typically present in infancy and childhood, although adult-onset forms also occur. Most LSDs have a progressive neurodegenerative clinical course, although symptoms in other organ systems are frequent. LSD-associated genes encode different lysosomal proteins, including lysosomal enzymes and lysosomal membrane proteins. The lysosome is the key cellular hub for macromolecule catabolism, recycling and signalling, and defects that impair any of these functions cause the accumulation of undigested or partially digested macromolecules in lysosomes (that is, 'storage') or impair the transport of molecules, which can result in cellular damage. Consequently, the cellular pathogenesis of these diseases is complex and is currently incompletely understood. Several LSDs can be treated with approved, disease-specific therapies that are mostly based on enzyme replacement. However, small-molecule therapies, including substrate reduction and chaperone therapies, have also been developed and are approved for some LSDs, whereas gene therapy and genome editing are at advanced preclinical stages and, for a few disorders, have already progressed to the clinic.


Assuntos
Doenças por Armazenamento dos Lisossomos/genética , Doença de Fabry/epidemiologia , Doença de Fabry/genética , Doença de Gaucher/epidemiologia , Doença de Gaucher/genética , Doença de Depósito de Glicogênio Tipo II/epidemiologia , Doença de Depósito de Glicogênio Tipo II/genética , Humanos , Leucodistrofia Metacromática/epidemiologia , Leucodistrofia Metacromática/genética , Doenças por Armazenamento dos Lisossomos/epidemiologia , Proteínas/análise
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