Farber disease (FD) and spinal muscular atrophy with progressive myoclonic epilepsy (SMA-PME) are ultra-rare lysosomal storage disorders caused by deficient acid ceramidase (ACDase) activity. Although both conditions are caused by mutations in the ASAH1 gene, clinical presentations differ considerably. FD patients usually die in childhood, while SMA-PME patients can live until adulthood. There is no treatment for FD or SMA-PME. Hematopoietic stem cell transplantation (HSCT) and gene therapy strategies for the treatment of ACDase deficiency are being investigated. We have previously generated and characterized mouse models of both FD and SMA-PME that recapitulate the symptoms described in patients. Here, we show that HSCT improves lifespan, behavior, hematopoietic system anomalies, and plasma cytokine levels and significantly reduces histiocytic infiltration and ceramide accumulation throughout the tissues investigated, including the CNS, in both models of ACDase-deficient mice. HSCT was also successful in preventing lesion development and significant demyelination of the spinal cord seen in SMA-PME mice. Importantly, we note that only early and generally pre-symptomatic treatment was effective, and kidney impairment was not improved in either model.

• Klíčová slova
• Farber disease, HSCT, Lysosomal storage disorders, central nervous system, ceramides, spinal muscular atrophy with progressive myoclonic epilepsy (SMA-PME),
• MeSH
• ceramidy metabolismus   MeSH
• Farberova nemoc * terapie   genetika   MeSH
• kyselá ceramidasa * genetika   metabolismus   MeSH
• lidé MeSH
• mícha metabolismus   patologie   MeSH
• modely nemocí na zvířatech MeSH
• myoklonické epilepsie progresivní genetika   terapie   metabolismus   MeSH
• myši knockoutované MeSH
• myši MeSH
• transplantace hematopoetických kmenových buněk * metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• Asah1 protein, mouse MeSH Prohlížeč
• ceramidy MeSH
• kyselá ceramidasa * MeSH

Acid ceramidase catalyzes the degradation of ceramide into sphingosine and a free fatty acid. Acid ceramidase deficiency results in lipid accumulation in many tissues and leads to the development of Farber disease (FD). Typical manifestations of classical FD include formation of subcutaneous nodules and joint contractures as well as the development of a hoarse voice. Healthy skin depends on a unique lipid profile to form a barrier that confers protection from pathogens, prevents excessive water loss, and mediates cell-cell communication. Ceramides comprise ~50% of total epidermis lipids and regulate cutaneous homeostasis and inflammation. Abnormal skin development including visual skin lesions has been reported in FD patients, but a detailed study of FD skin has not been performed. We conducted a pathophysiological study of the skin in our mouse model of FD. We observed altered lipid composition in FD skin dominated by accumulation of all studied ceramide species and buildup of abnormal storage structures affecting mainly the dermis. A deficiency of acid ceramidase activity also led to the activation of inflammatory IL-6/JAK/signal transducer and activator of transcription 3 and noncanonical NF-κB signaling pathways. Last, we report reduced proliferation of FD mouse fibroblasts and adipose-derived stem/stromal cells (ASC) along with impaired differentiation of ASCs into mature adipocytes.

• Klíčová slova
• Farber disease, acid ceramidase, adipogenesis, ceramides, macrophages, skin,
• MeSH
• adipogeneze MeSH
• ceramidy metabolismus   MeSH
• Farberova nemoc * MeSH
• kyselá ceramidasa genetika   MeSH
• modely nemocí na zvířatech MeSH
• myši MeSH
• zánět MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• ceramidy MeSH
• kyselá ceramidasa MeSH

Mucolipidosis type IV (MLIV) is a lysosomal storage disease exhibiting progressive intellectual disability, motor impairment, and premature death. There is currently no cure or corrective treatment. The disease results from mutations in the gene encoding mucolipin-1, a transient receptor potential channel believed to play a key role in lysosomal calcium egress. Loss of mucolipin-1 and subsequent defects lead to a host of cellular aberrations, including accumulation of glycosphingolipids (GSLs) in neurons and other cell types, microgliosis and, as reported here, cerebellar Purkinje cell loss. Several studies have demonstrated that N-butyldeoxynojirimycin (NB-DNJ, also known as miglustat), an inhibitor of the enzyme glucosylceramide synthase (GCS), successfully delays the onset of motor deficits, improves longevity, and rescues some of the cerebellar abnormalities (e.g., Purkinje cell death) seen in another lysosomal disease known as Niemann-Pick type C (NPC). Given the similarities in pathology between MLIV and NPC, we examined whether miglustat would be efficacious in ameliorating disease progression in MLIV. Using a full mucolipin-1 knockout mouse (Mcoln1-/-), we found that early miglustat treatment delays the onset and progression of motor deficits, delays cerebellar Purkinje cell loss, and reduces cerebellar microgliosis characteristic of MLIV disease. Quantitative mass spectrometry analyses provided new data on the GSL profiles of murine MLIV brain tissue and showed that miglustat partially restored the wild type profile of white matter enriched lipids. Collectively, our findings indicate that early miglustat treatment delays the progression of clinically relevant pathology in an MLIV mouse model, and therefore supports consideration of miglustat as a therapeutic agent for MLIV disease in humans.

• Klíčová slova
• Glycosphingolipids, Lysosomal storage disease, Miglustat, Mucolipidosis type IV, Mucolipin-1, Purkinje cells, Small molecule therapy,
• MeSH
• 1-deoxynojirimycin analogy a deriváty   terapeutické užití   MeSH
• CD antigeny metabolismus   MeSH
• glióza farmakoterapie   etiologie   MeSH
• inhibitory enzymů terapeutické užití   MeSH
• kationtové kanály TRP genetika   metabolismus   MeSH
• metabolismus lipidů účinky léků   genetika   MeSH
• modely nemocí na zvířatech MeSH
• mozeček patologie   MeSH
• mukolipidózy * komplikace   genetika   patologie   MeSH
• myši inbrední C57BL MeSH
• myši transgenní MeSH
• myši MeSH
• pátrací chování účinky léků   MeSH
• počet buněk MeSH
• pohybové poruchy farmakoterapie   etiologie   MeSH
• proteiny nervové tkáně metabolismus   MeSH
• psychomotorický výkon účinky léků   MeSH
• Purkyňovy buňky účinky léků   patologie   MeSH
• retina patologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• 1-deoxynojirimycin MeSH
• CD antigeny MeSH
• inhibitory enzymů MeSH
• kationtové kanály TRP MeSH
• Mcoln1 protein, mouse MeSH Prohlížeč
• miglustat MeSH Prohlížeč
• proteiny nervové tkáně MeSH

BACKGROUND: Prediagnostic steps in suspected metachromatic leukodystrophy (MLD) rely on clinical chemical methods other than enzyme assays. We report a new diagnostic method which evaluates changes in the spectrum of molecular types of sulfatides (3-O-sulfogalactosyl ceramides) in MLD urine. METHODS: The procedure allows isolation of urinary sulfatides by solid-phase extraction on DEAE-cellulose membranes, transportation of a dry membrane followed by elution and tandem mass spectrometry (MS/MS) analysis in the clinical laboratory. Major sulfatide isoforms are normalized to the least variable component of the spectrum, which is the indigenous C18:0 isoform. This procedure does not require the use of specific internal standards and minimizes errors caused by sample preparation and measurement. RESULTS: Urinary sulfatides were analyzed in a set of 21 samples from patients affected by sulfatidosis. The combined abundance of the five most elevated isoforms, C22:0, C22:0-OH, C24:0, C24:1-OH, and C24:0-OH sulfatides, was found to give the greatest distinction between MLD-affected patients and a control group. CONCLUSIONS: The method avoids transportation of liquid urine samples and generates stable membrane-bound sulfatide samples that can be stored at ambient temperature. MS/MS sulfatide profiling targeted on the most MLD-representative isoforms is simple with robust results and is suitable for screening.

• Klíčová slova
• ASA, CV, DEAE, DEAE-cellulose membrane, DUS, Diethylaminoethyl, Dry urinary samples, IPN, Isoforms, MLD, MS/MS, PTFE, Psap-d, S/N, SRM, Screening for metachromatic leukodystrophy, Tandem mass spectrometry, Urinary sulfatide, arylsulfatase A, coefficient of variation, dry urinary sample, isoform profile number (ratio of the sum of the major five isoforms and the C18:0 sulfatide), metachromatic leukodystrophy, polytetrafluoroethylene, prosaposin deficiency, selected reaction monitoring, signal to noise ratio, tandem mass spectrometry,
• MeSH
• DEAE-celulosa MeSH
• dítě MeSH
• extrakce na pevné fázi MeSH
• kojenec MeSH
• lidé středního věku MeSH
• lidé MeSH
• membrány umělé MeSH
• metachromatická leukodystrofie diagnóza   moč   MeSH
• mladiství MeSH
• odběr biologického vzorku normy   MeSH
• předškolní dítě MeSH
• referenční standardy MeSH
• studie případů a kontrol MeSH
• sulfoglykosfingolipidy moč   MeSH
• tandemová hmotnostní spektrometrie MeSH
• vysoušení MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• DEAE-celulosa MeSH
• membrány umělé MeSH
• sulfoglykosfingolipidy MeSH

The aim of this retrospective study was to determine the prevalence of lysosomal storage disorders (LSDs) in the Czech Republic. The data on cases diagnosed between 1975 and 2008 were collected and analyzed. The overall prevalence of LSDs in the Czech population (12.25 per 100,000) is comparable to that reported for the countries with well-established and advanced diagnostics of LSDs such as the Netherlands (14 per 100,000), Australia (12.9 per 100,000) and Italy (12.1 per 100,000). Relatively higher prevalence of LSDs was reported in the north of Portugal (25 per 100,000). Thirty-four different LSDs were diagnosed in a total of 478 individuals. Gaucher disease was the most frequent LSD with a birth prevalence of 1.13 per 100,000 births. The most frequent LSD groups were lipidoses, mucopolysaccharidoses, and neuronal ceroid lipofuscinoses, with combined prevalences of 5.0, 3.72, and 2.29 per 100,000 live births, respectively. Glycoproteinoses (0.57 per 100,000 live births), glycogenosis type II (0.37), and mucolipidoses (0.31) rarely occur in the Czech population, and a range of other LSDs have not been detected at all over the past three decades. Knowledge of the birth prevalence and carrier frequency of particular disorders is important in genetic counselling for calculation of the risk for the disorder in the other members of affected families. Earlier diagnosis of these disorders will permit timely intervention and may also result in lowering of the number of newborns with LSDs.

• MeSH
• genetická predispozice k nemoci epidemiologie   MeSH
• genetické poradenství MeSH
• heterozygot MeSH
• lidé MeSH
• lyzozomální nemoci z ukládání epidemiologie   genetika   MeSH
• novorozenec MeSH
• prevalence MeSH
• retrospektivní studie MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• novorozenec MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Geografické názvy
• Austrálie epidemiologie   MeSH
• Česká republika epidemiologie   MeSH
• Itálie epidemiologie   MeSH
• Nizozemsko epidemiologie   MeSH
• Portugalsko epidemiologie   MeSH