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1.
Int J Mol Sci ; 24(22)2023 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-38003660

RESUMO

Calcified aortic valve disease in its final stage leads to aortic valve stenosis, limiting cardiac function. To date, surgical intervention is the only option for treating calcific aortic valve stenosis. This study combined controlled drug delivery by nanoparticles (NPs) and active targeting by antibody conjugation. The chelating agent diethylenetriaminepentaacetic acid (DTPA) was covalently bound to human serum albumin (HSA)-based NP, and the NP surface was modified using conjugating antibodies (anti-elastin or isotype IgG control). Calcification was induced ex vivo in porcine aortic valves by preincubation in an osteogenic medium containing 2.5 mM sodium phosphate for five days. Valve calcifications mainly consisted of basic calcium phosphate crystals. Calcifications were effectively resolved by adding 1-5 mg DTPA/mL medium. Incubation with pure DTPA, however, was associated with a loss of cellular viability. Reversal of calcifications was also achieved with DTPA-coupled anti-elastin-targeted NPs containing 1 mg DTPA equivalent. The addition of these NPs to the conditioned media resulted in significant regression of the valve calcifications compared to that in the IgG-NP control without affecting cellular viability. These results represent a step further toward the development of targeted nanoparticular formulations to dissolve aortic valve calcifications.


Assuntos
Estenose da Valva Aórtica , Nanopartículas , Humanos , Animais , Suínos , Elastina/metabolismo , Estenose da Valva Aórtica/tratamento farmacológico , Estenose da Valva Aórtica/metabolismo , Valva Aórtica/metabolismo , Ácido Pentético , Imunoglobulina G/metabolismo
2.
Int J Mol Sci ; 23(2)2022 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-35055176

RESUMO

Cytokines and their receptors have a vital function in regulating various processes such as immune function, inflammation, haematopoiesis, cell growth and differentiation. The interaction between a cytokine and its specific receptor triggers intracellular signalling cascades that lead to altered gene expression in the target cell and consequent changes in its proliferation, differentiation, or activation. In this review, we highlight the role of the soluble type I cytokine receptor CRLF1 (cytokine receptor-like factor-1) and the Interleukin (IL)-6 cytokine CLCF1 (cardiotrophin-like cytokine factor 1) during development in physiological and pathological conditions with particular emphasis on Crisponi/cold-induced sweating syndrome (CS/CISS) and discuss new insights, challenges and possibilities arising from recent studies.


Assuntos
Citocinas/metabolismo , Deformidades Congênitas da Mão/genética , Hiperidrose/genética , Receptores de Citocinas/metabolismo , Trismo/congênito , Animais , Morte Súbita , Modelos Animais de Doenças , Fácies , Desenvolvimento Humano , Humanos , Transdução de Sinais , Trismo/genética
3.
Arterioscler Thromb Vasc Biol ; 41(1): 35-47, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33176451

RESUMO

Arterial calcification is a common phenomenon in the elderly, in patients with atherosclerosis or renal failure and in diabetes. However, when present in very young individuals, it is likely to be associated with an underlying hereditary disorder of arterial calcification. Here, we present an overview of the few monogenic disorders presenting with early-onset cardiovascular calcification. These disorders can be classified according to the function of the respective disease gene into (1) disorders caused by an altered purine and phosphate/pyrophosphate metabolism, (2) interferonopathies, and (3) Gaucher disease. The finding of arterial calcification in early life should alert the clinician and prompt further genetic work-up to define the underlying genetic defect, to establish the correct diagnosis, and to enable appropriate therapy.


Assuntos
Doenças da Aorta/genética , Artérias/metabolismo , Hipoplasia do Esmalte Dentário/genética , Hereditariedade , Erros Inatos do Metabolismo/genética , Metacarpo/anormalidades , Doenças Musculares/genética , Odontodisplasia/genética , Osteogênese/genética , Osteoporose/genética , Calcificação Vascular/genética , Animais , Doenças da Aorta/complicações , Doenças da Aorta/metabolismo , Artérias/patologia , Hipoplasia do Esmalte Dentário/complicações , Hipoplasia do Esmalte Dentário/metabolismo , Doença de Gaucher/complicações , Doença de Gaucher/genética , Doença de Gaucher/metabolismo , Predisposição Genética para Doença , Humanos , Erros Inatos do Metabolismo/complicações , Erros Inatos do Metabolismo/metabolismo , Metacarpo/metabolismo , Doenças Musculares/complicações , Doenças Musculares/metabolismo , Odontodisplasia/complicações , Odontodisplasia/metabolismo , Osteoporose/complicações , Osteoporose/metabolismo , Fenótipo , Medição de Risco , Fatores de Risco , Calcificação Vascular/complicações , Calcificação Vascular/metabolismo , Calcificação Vascular/patologia
4.
Stem Cell Res ; 46: 101855, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32512309

RESUMO

Crisponi syndrome/cold-induced sweating syndrome type 2 (CS/CISS2) is a rare disease with severe dysfunctions of thermoregulatory processes. CS/CISS2 individuals suffer from recurrent episodes of hyperthermia in the neonatal period and paradoxical sweating at cold ambient temperatures in adolescence. Variants in CLCF1 (cardiotrophin-like-cytokine 1) cause CS/CISS2. Here, we summarize the generation of three clones of one stem cell line (iPSC) of a CS/CISS2 individual carrying the CLCF1 variant c.321C>G on both alleles. These patient derived iPSC clones show a normal karyotype, several pluripotency markers, and the ability to differentiate into the three germ layers.


Assuntos
Células-Tronco Pluripotentes Induzidas , Adolescente , Reprogramação Celular , Células Clonais , Morte Súbita , Fácies , Fibroblastos , Deformidades Congênitas da Mão , Humanos , Hiperidrose , Recém-Nascido , Receptores de Citocinas/genética , Trismo/congênito
5.
Stem Cell Res ; 46: 101820, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32492556

RESUMO

Cytokine receptor like factor 1 (CRLF1) is the gene implicated, when mutated, in Crisponi syndrome/cold-induced sweating syndrome type 1 (CS/CISS1). Here, we report the establishment of induced pluripotent stem cell lines (iPSCs) from fibroblasts of a Turkish CS/CISS1 individual with a homozygous variant in CRLF1 (c.708_709delinsT; p.[Pro238Argfs*6]). This variant is the most frequent variant associated to CS/CISS1 in the Turkish population. These patient derived iPSC lines show all pluripotency markers, a normal karyotype and the ability to differentiate into the three germ layers.


Assuntos
Deformidades Congênitas da Mão , Hiperidrose , Células-Tronco Pluripotentes Induzidas , Fácies , Humanos , Sudorese
6.
Clin Genet ; 97(1): 209-221, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31497877

RESUMO

Crisponi/cold-induced sweating syndrome (CS/CISS) is an autosomal recessive disease characterized by hyperthermia, camptodactyly, feeding and respiratory difficulties often leading to sudden death in the neonatal period. The affected individuals who survived the first critical years of life, develop cold-induced sweating and scoliosis in early childhood. The disease is caused by variants in the CRLF1 or in the CLCF1 gene. Both proteins form a heterodimeric complex that acts on cells expressing the ciliary neurotrophic factor receptor (CNTFR). CS/CISS belongs to the family of "CNTFR-related disorders" showing a similar clinical phenotype. Recently, variants in other genes, including KLHL7, NALCN, MAGEL2 and SCN2A, previously linked to other diseases, have been associated with a CS/CISS-like phenotype. Therefore, retinitis pigmentosa and Bohring-Optiz syndrome-like (KLHL7), Congenital contractures of the limbs and face, hypotonia, and developmental delay syndrome (NALCN), Chitayat-Hall/Schaaf-Yang syndrome (MAGEL2), and early infantile epileptic encephalopathy-11 syndrome (SCN2A) all share an overlapping phenotype with CS/CISS, especially in the neonatal period. This review aims to summarize the existing literature on CS/CISS, focusing on the current state of differential diagnosis, pathogenesis and treatment concepts in order to achieve an accurate and rapid diagnosis. This will improve patient management and enable specific treatments for the affected individuals.


Assuntos
Craniossinostoses/diagnóstico , Citocinas/genética , Deformidades Congênitas da Mão/diagnóstico , Hiperidrose/diagnóstico , Deficiência Intelectual/diagnóstico , Receptores de Citocinas/genética , Trismo/congênito , Subunidade alfa do Receptor do Fator Neutrófico Ciliar/genética , Craniossinostoses/genética , Craniossinostoses/patologia , Morte Súbita/patologia , Diagnóstico Diferencial , Fácies , Deformidades Congênitas da Mão/patologia , Deformidades Congênitas da Mão/terapia , Humanos , Hiperidrose/patologia , Hiperidrose/terapia , Deficiência Intelectual/genética , Deficiência Intelectual/patologia , Retinose Pigmentar/diagnóstico , Retinose Pigmentar/genética , Retinose Pigmentar/patologia , Escoliose/diagnóstico , Trismo/diagnóstico , Trismo/patologia , Trismo/terapia
7.
Clin Genet ; 95(5): 607-614, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30859550

RESUMO

Crisponi/cold-induced sweating syndrome (CS/CISS) is a rare autosomal recessive disorder characterized by a complex phenotype (hyperthermia and feeding difficulties in the neonatal period, followed by scoliosis and paradoxical sweating induced by cold since early childhood) and a high neonatal lethality. CS/CISS is a genetically heterogeneous disorder caused by mutations in CRLF1 (CS/CISS1), CLCF1 (CS/CISS2) and KLHL7 (CS/CISS-like). Here, a whole exome sequencing approach in individuals with CS/CISS-like phenotype with unknown molecular defect revealed unpredicted alternative diagnoses. This approach identified putative pathogenic variations in NALCN, MAGEL2 and SCN2A. They were already found implicated in the pathogenesis of other syndromes, respectively the congenital contractures of the limbs and face, hypotonia, and developmental delay syndrome, the Schaaf-Yang syndrome, and the early infantile epileptic encephalopathy-11 syndrome. These results suggest a high neonatal phenotypic overlap among these disorders and will be very helpful for clinicians. Genetic analysis of these genes should be considered for those cases with a suspected CS/CISS during neonatal period who were tested as mutation negative in the known CS/CISS genes, because an expedited and corrected diagnosis can improve patient management and can provide a specific clinical follow-up.


Assuntos
Sequenciamento do Exoma , Deformidades Congênitas da Mão/diagnóstico , Deformidades Congênitas da Mão/genética , Hiperidrose/diagnóstico , Hiperidrose/genética , Trismo/congênito , Morte Súbita , Fácies , Feminino , Humanos , Lactente , Masculino , Linhagem , Fenótipo , Trismo/diagnóstico , Trismo/genética
8.
Exp Mol Med ; 50(10): 1-12, 2018 10 29.
Artigo em Inglês | MEDLINE | ID: mdl-30369595

RESUMO

Generalized arterial calcification of infancy (GACI) is associated with widespread arterial calcification and stenoses and is caused by mutations in ENPP1. ENPP1 encodes for ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1), which cleaves ATP to generate inorganic pyrophosphate (PPi) and adenosine monophosphate (AMP) extracellularly. The current study was designed to define the prevalence of arterial stenoses in GACI individuals and to identify the mechanism through which ENPP1 deficiency causes intimal proliferation. Furthermore, we aimed to effectively prevent and treat neointima formation in an animal model of GACI through the systemic administration of recombinant human (rh)ENPP1-Fc protein. Based on a literature review, we report that arterial stenoses are present in at least 72.4% of GACI cases. We evaluated the effect of rhENPP1-Fc on ENPP1-silenced human vascular smooth muscle cells (VSMCs) and on induced intimal proliferation in Enpp1-deficient ttw/ttw mice treated with carotid ligation. We demonstrate that silencing ENPP1 in VSMCs resulted in a tenfold increase in proliferation relative to that of cells transfected with negative control siRNA. The addition of rhENPP1-Fc, AMP or adenosine restored the silenced ENPP1-associated proliferation. In contrast, neither PPi nor etidronate, a current off-label treatment for GACI, had an effect on VSMC proliferation. Furthermore, subcutaneous rhENPP1-Fc protein replacement was effective in preventing and treating intimal hyperplasia induced by carotid ligation in an animal model of GACI. We conclude that ENPP1 inhibits neointima formation by generating  AMP. RhENPP1-Fc may serve as an approach for the effective prevention and treatment of arterial stenoses in GACI.


Assuntos
Monofosfato de Adenosina/biossíntese , Fragmentos Fc das Imunoglobulinas/farmacologia , Neointima/metabolismo , Neointima/patologia , Diester Fosfórico Hidrolases/farmacologia , Pirofosfatases/farmacologia , Proteínas Recombinantes de Fusão/farmacologia , Calcificação Vascular/metabolismo , Calcificação Vascular/patologia , Trifosfato de Adenosina/sangue , Trifosfato de Adenosina/metabolismo , Animais , Proliferação de Células/efeitos dos fármacos , Modelos Animais de Doenças , Feminino , Técnicas de Inativação de Genes , Humanos , Fragmentos Fc das Imunoglobulinas/genética , Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Pluripotentes Induzidas/metabolismo , Masculino , Camundongos , Camundongos Knockout , Modelos Biológicos , Miócitos de Músculo Liso/citologia , Miócitos de Músculo Liso/metabolismo , Neointima/etiologia , Neointima/prevenção & controle , Diester Fosfórico Hidrolases/genética , Pirofosfatases/genética , RNA Interferente Pequeno/genética , Ratos , Proteínas Recombinantes de Fusão/genética , Calcificação Vascular/tratamento farmacológico , Calcificação Vascular/etiologia
10.
J Interferon Cytokine Res ; 37(5): 214-219, 2017 05.
Artigo em Inglês | MEDLINE | ID: mdl-28475458

RESUMO

In 1973, Singleton and Merten described a new syndrome in 2 female probands with aortic and cardiac valve calcifications, early loss of secondary dentition, and widened medullary cavities of the phalanges. In 1984, Aicardi and Goutières defined a phenotype resembling congenital viral infection with basal ganglia calcification and increased protein content in the cerebrospinal fluid. Between 2006 and 2012, mutations in 6 different genes were described to be associated with Aicardi-Goutières syndrome, specifically-TREX1, RNASEH2A, RNASEH2B, RNASEH2C, ADAR, and SAMHD1. More recently, mutations in IFIH1 were reported in a variety of neuroimmunological phenotypes, including Aicardi-Goutières syndrome, while a specific Arg822Gln mutation in IFIH1 was described in 3 discrete families with Singleton-Merten syndrome (SMS). IFIH1 encodes for melanoma differentiation-associated gene 5 (MDA5), and all mutations identified to date have been associated with an enhanced interferon response in affected individuals. In this study, we present a male child demonstrating recurrent febrile episodes, spasticity, and basal ganglia calcification suggestive of Aicardi-Goutières syndrome, who carries the same Arg822Gln mutation in IFIH1 previously associated with SMS. We conclude that both diseases are part of the interferonopathy grouping and that the Arg822Gln mutation in IFIH1 can cause a spectrum of disease, including neurological involvement.


Assuntos
Doenças da Aorta/imunologia , Hipoplasia do Esmalte Dentário/imunologia , Inflamação/imunologia , Interferon Tipo I/imunologia , Helicase IFIH1 Induzida por Interferon/imunologia , Metacarpo/anormalidades , Doenças Musculares/imunologia , Odontodisplasia/imunologia , Osteoporose/imunologia , Calcificação Vascular/imunologia , Doenças da Aorta/genética , Criança , Hipoplasia do Esmalte Dentário/genética , Humanos , Helicase IFIH1 Induzida por Interferon/genética , Masculino , Metacarpo/imunologia , Doenças Musculares/genética , Mutação , Odontodisplasia/genética , Osteoporose/genética , Calcificação Vascular/genética
11.
Am J Hum Genet ; 99(1): 236-45, 2016 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-27392078

RESUMO

Crisponi syndrome (CS)/cold-induced sweating syndrome type 1 (CISS1) is a very rare autosomal-recessive disorder characterized by a complex phenotype with high neonatal lethality, associated with the following main clinical features: hyperthermia and feeding difficulties in the neonatal period, scoliosis, and paradoxical sweating induced by cold since early childhood. CS/CISS1 can be caused by mutations in cytokine receptor-like factor 1 (CRLF1). However, the physiopathological role of CRLF1 is still poorly understood. A subset of CS/CISS1 cases remain yet genetically unexplained after CRLF1 sequencing. In five of them, exome sequencing and targeted Sanger sequencing identified four homozygous disease-causing mutations in kelch-like family member 7 (KLHL7), affecting the Kelch domains of the protein. KLHL7 encodes a BTB-Kelch-related protein involved in the ubiquitination of target proteins for proteasome-mediated degradation. Mono-allelic substitutions in other domains of KLHL7 have been reported in three families affected by a late-onset form of autosomal-dominant retinitis pigmentosa. Retinitis pigmentosa was also present in two surviving children reported here carrying bi-allelic KLHL7 mutations. KLHL7 mutations are thus associated with a more severe phenotype in recessive than in dominant cases. Although these data further support the pathogenic role of KLHL7 mutations in a CS/CISS1-like phenotype, they do not explain all their clinical manifestations and highlight the high phenotypic heterogeneity associated with mutations in KLHL7.


Assuntos
Alelos , Autoantígenos/genética , Deformidades Congênitas da Mão/complicações , Deformidades Congênitas da Mão/genética , Hiperidrose/complicações , Hiperidrose/genética , Mutação , Retinose Pigmentar/complicações , Retinose Pigmentar/genética , Trismo/congênito , Sequência de Aminoácidos , Autoantígenos/química , Criança , Pré-Escolar , Morte Súbita , Fácies , Feminino , Humanos , Lactente , Masculino , Modelos Moleculares , Linhagem , Fenótipo , Síndrome , Trismo/complicações , Trismo/genética
12.
Hum Mutat ; 37(11): 1190-1201, 2016 11.
Artigo em Inglês | MEDLINE | ID: mdl-27467858

RESUMO

Ectonucleotide pyrophosphatase/phosphodiesterase family member 1 (E-NPP1), encoded by ENPP1, is a plasma membrane protein that generates inorganic pyrophosphate (PPi ), a physiologic inhibitor of hydroxyapatite formation. In humans, variants in ENPP1 are associated with generalized arterial calcification of infancy, an autosomal-recessive condition causing premature onset of arterial calcification and intimal proliferation resulting in stenoses. ENPP1 variants also cause pseudoxanthoma elasticum characterized by ectopic calcification of soft connective tissues. To determine the functional impact of ENPP1 missense variants, we analyzed 13 putative pathogenic variants in vitro regarding their functional properties, that is, activity, localization, and PPi generation. Transfection of eight of the 13 variants led to complete loss of NPP activity, whereas four mutants (c.1412A > G, p.Tyr471Cys; c.1510A > C, p.Ser504Arg; c.1976A > G, p.Tyr659Cys; c.2330A > G, p.His777Arg) showed residual activity compared with wild-type E-NPP1. One putative pathologic variant (c.2462 G > A, p.Arg821His) showed normal activity. The five mutants with normal or residual E-NPP1 enzyme activity were still able to generate PPi and localized in the plasma membrane. In this study, we identified a functional ENPP1 polymorphism, which was expected to be pathogenic till now. Furthermore, we identified four mutants (p.Tyr471Cys, p.Ser504Arg, p.Tyr659Cys, p.His777Arg) with residual E-NPP1 function, which would be potential therapeutical targets for conformational-stabilizing agents.


Assuntos
Mutação de Sentido Incorreto , Diester Fosfórico Hidrolases/genética , Diester Fosfórico Hidrolases/metabolismo , Pirofosfatases/genética , Pirofosfatases/metabolismo , Calcificação Vascular/genética , Animais , Células COS , Membrana Celular/metabolismo , Chlorocebus aethiops , Difosfatos/metabolismo , Regulação para Baixo , Células HEK293 , Humanos , Mutagênese Sítio-Dirigida
13.
J Cell Mol Med ; 20(8): 1523-33, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27061115

RESUMO

The rare inborn cblF defect of cobalamin metabolism is caused by mutations in the limb region 1 (LMBR1) domain containing 1 gene (LMBRD1). This defect is characterized by massive accumulation of free cobalamin in lysosomes and loss of mitochondrial succinyl-CoA synthesis and cytosolic methionine synthesis. Affected children suffer from heart defects, developmental delay and megaloblastic anemia. LMBRD1 encodes for LMBD1, a predicted lysosomal cobalamin transport protein. In this study, we determine the physiological function of LMBRD1 during embryogenesis by generating Lmbrd1 deficient mice using the Cre/LoxP system. Complete loss of Lmbrd1 function is accompanied by early embryonic death in mice. Whole mount in situ hybridization studies against bone morphogenetic protein 4 and Nodal show that initial formation of the proximal-distal axis is unaffected in early embryonic stages whereas the initiation of gastrulation is disturbed shown by the expression pattern of even skipped homeotic gene 1 and fibroblast growth factor 8 in Lmbrd1 deficient mice. We conclude that intact function of LMBD1 is essential for the initiation of gastrulation.


Assuntos
Gastrulação , Proteínas de Transporte Nucleocitoplasmático/metabolismo , Animais , Biomarcadores/metabolismo , Perda do Embrião/genética , Perda do Embrião/patologia , Embrião de Mamíferos/metabolismo , Embrião de Mamíferos/patologia , Feminino , Marcação de Genes , Fator 3-beta Nuclear de Hepatócito/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Células-Tronco Embrionárias Murinas/metabolismo , Transfecção
14.
Cytokine Growth Factor Rev ; 29: 101-7, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-26993858

RESUMO

Type I interferonopathies are a relatively new class of inherited autoimmune disorders associated with an inborn elevated interferon response. Activation of cytosolic receptors which recognize viral double stranded RNA including the RIG-I (retinoic acid-inducible gene I) like receptors RIG-I and MDA5 (melanoma differentiation-associated gene 5) has been shown to induce the transcription of type I interferon genes. Within recent years, with the help of next generation sequencing techniques in syndromic families, mutations in the genes encoding for RIG-I and MDA5 have been identified to cause rare diseases including Aicardi-Goutières syndrome, Systemic Lupus Erythematosus in certain individuals as well as classic and atypical Singleton-Merten syndrome. Patients carrying mono-allelic mutations in MDA5 and RIG-I show constitutive activation of the RIG-I receptors and downstream signalling associated with increased type I interferon production. Although differing in the degree of phenotypic expression and severity, the phenotype of these "novel" diseases shows a considerable overlap reflecting their common pathogenetic pathway.


Assuntos
Doenças Autoimunes/genética , Proteína DEAD-box 58/genética , Interferon Tipo I/genética , Mutação , Transdução de Sinais/genética , Animais , Doenças Autoimunes/imunologia , Doenças Autoimunes/patologia , Proteína DEAD-box 58/imunologia , Humanos , Interferon Tipo I/imunologia , Helicase IFIH1 Induzida por Interferon/genética , Helicase IFIH1 Induzida por Interferon/imunologia , Receptores Imunológicos , Transdução de Sinais/imunologia
15.
Am J Hum Genet ; 96(2): 275-82, 2015 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-25620204

RESUMO

Singleton-Merten syndrome (SMS) is an infrequently described autosomal-dominant disorder characterized by early and extreme aortic and valvular calcification, dental anomalies (early-onset periodontitis and root resorption), osteopenia, and acro-osteolysis. To determine the molecular etiology of this disease, we performed whole-exome sequencing and targeted Sanger sequencing. We identified a common missense mutation, c.2465G>A (p.Arg822Gln), in interferon induced with helicase C domain 1 (IFIH1, encoding melanoma differentiation-associated protein 5 [MDA5]) in four SMS subjects from two families and a simplex case. IFIH1 has been linked to a number of autoimmune disorders, including Aicardi-Goutières syndrome. Immunohistochemistry demonstrated the localization of MDA5 in all affected target tissues. In vitro functional analysis revealed that the IFIH1 c.2465G>A mutation enhanced MDA5 function in interferon beta induction. Interferon signature genes were upregulated in SMS individuals' blood and dental cells. Our data identify a gain-of-function IFIH1 mutation as causing SMS and leading to early arterial calcification and dental inflammation and resorption.


Assuntos
Doenças da Aorta/genética , RNA Helicases DEAD-box/genética , Hipoplasia do Esmalte Dentário/genética , Metacarpo/anormalidades , Modelos Moleculares , Doenças Musculares/genética , Odontodisplasia/genética , Osteoporose/genética , Fenótipo , Calcificação Vascular/genética , Sequência de Aminoácidos , Artérias/patologia , Sequência de Bases , Calcinose/genética , Calcinose/patologia , RNA Helicases DEAD-box/química , RNA Helicases DEAD-box/metabolismo , Exoma/genética , Genes Dominantes/genética , Humanos , Imuno-Histoquímica , Helicase IFIH1 Induzida por Interferon , Interferon beta/metabolismo , Dados de Sequência Molecular , Mutação de Sentido Incorreto/genética , Linhagem , Análise de Sequência de DNA , Anormalidades Dentárias/genética , Anormalidades Dentárias/patologia
16.
J Nutr Biochem ; 26(4): 327-36, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25595097

RESUMO

Lipid accumulation in macrophages contributes to atherosclerosis. Within macrophages, lipids are stored in lipid droplets (LDs); perilipin-2 and perilipin-3 are the main LD-associated proteins. Postprandial triglyceride (TG)-rich lipoproteins induce LD accumulation in macrophages. The role of postprandial lipoproteins in perilipin-2 and perilipin-3 regulation was studied. TG-rich lipoproteins (TRLs) induced the levels of intracellular TGs, LDs and perilipin-2 protein expression in THP-1 macrophages and in Apoe(-/-) mice bone-marrow-derived macrophages with low and high basal levels of TGs. Perilipin-3 was only synthesized in mice macrophages with low basal levels of TGs. The regulation was dependent on the fatty acid composition of the lipoproteins; monounsaturated and polyunsaturated fatty acids (PUFAs) more strongly attenuated these effects compared with saturated fatty acids. In THP-1 macrophages, immunofluorescence microscopy and freeze-fracture immunogold labeling indicated that the lipoproteins translocated perilipin-3 from the cytoplasm to the LD surface; only the lipoproteins that were rich in PUFAs suppressed this effect. Chemical inhibition showed that lipoproteins induced perilipin-2 protein expression through the peroxisome proliferator-activated nuclear receptor (PPAR) PPARα and PPARγ pathways. Overall, our data indicate that postprandial TRLs may be involved in atherosclerotic plaque formation through the regulation of perilipin-2 and perilipin-3 proteins in macrophages. Because the fatty acid composition of the lipoproteins is dependent on the type of fat consumed, the ingestion of olive oil, which is rich in monounsaturated fatty acids, and fish oil, which is rich in omega-3 fatty acids, can be considered a good nutritional strategy to reduce the risk of atherosclerosis by LD-associated proteins decrease.


Assuntos
Proteínas de Transporte/metabolismo , Lipoproteínas/sangue , Macrófagos/metabolismo , Proteínas de Membrana/metabolismo , Período Pós-Prandial , Triglicerídeos/sangue , Adulto , Animais , Manteiga , Proteínas de Transporte/genética , Linhagem Celular , Estudos Cross-Over , Ácidos Graxos/administração & dosagem , Ácidos Graxos/sangue , Ácidos Graxos Monoinsaturados/administração & dosagem , Ácidos Graxos Monoinsaturados/sangue , Ácidos Graxos Insaturados/administração & dosagem , Ácidos Graxos Insaturados/sangue , Óleos de Peixe/administração & dosagem , Regulação da Expressão Gênica , Humanos , Masculino , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Azeite de Oliva/administração & dosagem , PPAR alfa/genética , PPAR alfa/metabolismo , PPAR gama/genética , PPAR gama/metabolismo , Perilipina-2 , Perilipina-3 , Adulto Jovem
17.
Hum Mutat ; 35(4): 424-33, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24488861

RESUMO

Crisponi syndrome (CS) and cold-induced sweating syndrome type 1 (CISS1) share clinical characteristics, such as dysmorphic features, muscle contractions, scoliosis, and cold-induced sweating, with CS patients showing a severe clinical course in infancy involving hyperthermia associated with death in most cases in the first years of life. To date, 24 distinct CRLF1 mutations have been found either in homozygosity or in compound heterozygosity in CS/CISS1 patients, with the highest prevalence in Sardinia, Turkey, and Spain. By reporting 11 novel CRLF1 mutations, here we expand the mutational spectrum of CRLF1 in the CS/CISS1 syndrome to a total of 35 variants and present an overview of the different molecular and clinical features of all of them. To catalog all the 35 mutations, we created a CRLF1 mutations database, based on the Leiden Open (source) Variation Database (LOVD) system (https://grenada.lumc.nl/LOVD2/mendelian_genes/variants). Overall, the available functional and clinical data support the fact that both syndromes actually represent manifestations of the same autosomal-recessive disorder caused by mutations in the CRLF1 gene. Therefore, we propose to rename the two overlapping entities with the broader term of Crisponi/CISS1 syndrome.


Assuntos
Morte Súbita/patologia , Febre/genética , Febre/patologia , Deformidades Congênitas da Mão/genética , Deformidades Congênitas da Mão/patologia , Mutação , Receptores de Citocinas/genética , Trismo/congênito , Criança , Pré-Escolar , Subunidade alfa do Receptor do Fator Neutrófico Ciliar/genética , Bases de Dados Genéticas , Morte Súbita/epidemiologia , Fácies , Feminino , Febre/epidemiologia , Variação Genética , Deformidades Congênitas da Mão/epidemiologia , Humanos , Hiperidrose , Masculino , Contração Muscular/genética , Reação em Cadeia da Polimerase , Trismo/epidemiologia , Trismo/genética , Trismo/patologia
18.
Nat Genet ; 44(10): 1152-5, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22922874

RESUMO

Inherited disorders of vitamin B12 (cobalamin) have provided important clues to how this vitamin, which is essential for hematological and neurological function, is transported and metabolized. We describe a new disease that results in failure to release vitamin B12 from lysosomes, which mimics the cblF defect caused by LMBRD1 mutations. Using microcell-mediated chromosome transfer and exome sequencing, we identified causal mutations in ABCD4, a gene that codes for an ABC transporter, which was previously thought to have peroxisomal localization and function. Our results show that ABCD4 colocalizes with the lysosomal proteins LAMP1 and LMBD1, the latter of which is deficient in the cblF defect. Furthermore, we show that mutations altering the putative ATPase domain of ABCD4 affect its function, suggesting that the ATPase activity of ABCD4 may be involved in intracellular processing of vitamin B12.


Assuntos
Transportadores de Cassetes de Ligação de ATP/genética , Anormalidades Múltiplas/genética , Erros Inatos do Metabolismo/genética , Mutação , Vitamina B 12/metabolismo , Transportadores de Cassetes de Ligação de ATP/metabolismo , Anormalidades Múltiplas/enzimologia , Estudos de Casos e Controles , Células Cultivadas , Análise Mutacional de DNA , Fibroblastos/metabolismo , Expressão Gênica , Genes Recessivos , Estudos de Associação Genética , Humanos , Recém-Nascido , Proteínas de Membrana Lisossomal/metabolismo , Erros Inatos do Metabolismo/enzimologia , Proteínas de Transporte Nucleocitoplasmático/metabolismo , Estrutura Terciária de Proteína , Transporte Proteico
19.
Cold Spring Harb Protoc ; 2011(5): pdb.prot5612, 2011 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-21536764

RESUMO

INTRODUCTION: The human monocytic leukemia cell line THP-1 is a widely used model for investigating monocyte and macrophage biology. Successful transfection of THP-1 monocytes with subsequent phorbol 12-myristate 13-acetate (PMA)-induced differentiation into macrophages is not a trivial matter, because according to previous transfection protocols, cell viability is lost almost completely within 24 h of PMA treatment following transfection. This protocol constitutes an optimized version of a previously published protocol by our group. It describes a procedure for transfecting premature THP-1 macrophages, which subsequently can be further differentiated into mature macrophages by PMA without a loss of cell viability. Transfection of THP-1 cells with plasmids or small interfering RNA (siRNA) is achieved by electroporation using the Lonza Nucleofector technology (Basel, Switzerland). This technique allows for the efficient nonviral delivery of plasmids, DNA, RNA, or siRNA into primary cells or cell lines even if the cells are not or are only slowly proliferating. Such cells are usually rather difficult to transfect by nonviral approaches. This means that only viral approaches would be left, which are expensive and labor-intensive and require laboratories complying with the respective safety regulations. The protocol described here is an efficient and convenient alternative.


Assuntos
Macrófagos/fisiologia , Transfecção/métodos , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Eletroporação/métodos , Humanos , Plasmídeos/metabolismo , RNA Interferente Pequeno/metabolismo , Acetato de Tetradecanoilforbol/metabolismo
20.
Atherosclerosis ; 217(2): 371-8, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21612780

RESUMO

OBJECTIVE: The scavenger receptor SR-PSOX/CXCL16, which is identical to the chemokine CXCL16, is thought to be involved in atherogenesis. However, the presence and function of SR-PSOX/CXCL16 in the endothelium of atherosclerotic arteries has not been substantiated. METHODS AND RESULTS: In rabbit aorta immunocytochemistry revealed SR-PSOX/CXCL16 primarily in the endothelium at sites predisposed to lesion formation, in the endothelium of early atherosclerotic lesions, and mainly in intimal macrophages of more developed lesions, indicating that SR-PSOX/CXCL16-expression shifts during atherogenesis. In addition to its function as scavenger receptor and chemokine, SR-PSOX mediated the adhesion of THP-1 monocytes to endothelial cells in vitro. Both THP-1 monocytes and endothelial cells express SR-PSOX/CXCL16, and THP-1 monocytes express CXCR6, the specific receptor for SR-PSOX/CXCL16. Anti-SR-PSOX/CXCL16 and anti-CXCR6 antibody block monocyte adhesion, showing that SR-PSOX/CXCL16-CXCR6 interaction mediates monocyte-endothelial cell adhesion. SR-PSOX/CXCL16 expression of endothelial cells is upregulated by pro-inflammatory cytokines, and is reversed by incubation with ciglitazone and lovastatin. CONCLUSIONS: We suggest that SR-PSOX/CXCL16 may promote the adhesion of monocytes to the endothelium during early atherogenesis and that accumulating cytokines enhance SR-PSOX/CXCL16-mediated adhesion by upregulating SR-PSOX/CXCL16 expression. Manipulation of SR-PSOX/CXCL16 expression with anti-inflammatory agents may be of therapeutic value.


Assuntos
Aterosclerose/imunologia , Adesão Celular , Quimiocinas CXC/metabolismo , Células Endoteliais da Veia Umbilical Humana/imunologia , Monócitos/imunologia , Receptores Depuradores/metabolismo , Animais , Anti-Inflamatórios/farmacologia , Adesão Celular/efeitos dos fármacos , Células Cultivadas , Quimiocina CXCL16 , Técnicas de Cocultura , Citocinas/metabolismo , Modelos Animais de Doenças , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Hiperlipidemias/complicações , Hiperlipidemias/genética , Hiperlipidemias/imunologia , Imuno-Histoquímica , Mediadores da Inflamação/metabolismo , Lovastatina/farmacologia , Macrófagos/imunologia , Monócitos/efeitos dos fármacos , Coelhos , Tiazolidinedionas/farmacologia
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