scholarly journals Molecular mechanisms of pathogenesis in a glycosphingolipid and a glycoprotein storage disease

2010 ◽  
Vol 38 (6) ◽  
pp. 1453-1457 ◽  
Author(s):  
Alessandra d'Azzo ◽  
Erik Bonten
Keyword(s):  
Lysosomal Enzymes ◽  
Molecular Mechanisms ◽  
Storage Disease ◽  
Hydrolytic Enzymes ◽  
Lysosomal Storage ◽  
Serine Carboxypeptidase ◽  
Primary Defect ◽  
Apparent Lack ◽  
Storage Diseases ◽  
Lysosomal System

The lysosomal system comprises a specialized network of organelles crucial for the sorting, digestion, recycling and secretion of cellular components. With their content of hydrolytic enzymes, lysosomes regulate the degradation of a multitude of substrates that reach these organelles via the biosynthetic or the endocytic route. Gene defects that affect one or more of these hydrolases lead to LSDs (lysosomal storage diseases). This underscores the apparent lack of redundancy of these enzymes and the importance of the lysosomal system in cell and tissue homoeostasis. Some of the lysosomal enzymes may form multiprotein complexes, which usually work synergistically on substrates and, in this configuration, may respond more efficiently to changes in substrate load and composition. A well-characterized lysosomal multienzyme complex is the one comprising the glycosidases β-gal (β-galactosidase) and NEU1 (neuramidase-1), and of the serine carboxypeptidase PPCA (protective protein/cathepsin A). Three neurodegenerative LSDs are caused by either single or combined deficiency of these lysosomal enzymes. Sialidosis (NEU1 deficiency) and galactosialidosis (combined NEU1 and β-gal deficiency, secondary to a primary defect of PPCA) belong to the glycoprotein storage diseases, whereas GM1-gangliosidosis (β-gal deficiency) is a glycosphingolipid storage disease. Identification of novel molecular pathways that are deregulated because of loss of enzyme activity and/or accumulation of specific metabolites in various cell types has shed light on mechanisms of disease pathogenesis and may pave the way for future development of new therapies for these LSDs.

Development and Validation of Leukocyte Enzyme Activity Assay by LC-MS/MS for Diagnosis of Krabbe Disease and Other Lysosomal Storage Diseases

2020 ◽  
Vol 154 (Supplement_1) ◽  
pp. S16-S16
Author(s):  
Hsuan-Chieh Liao ◽  
Laura Mitchell ◽  
Katerina Sadilkova ◽  
Jane Dickerson ◽  
Rhona Jack ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Liquid Chromatography ◽  
Lysosomal Enzymes ◽  
Internal Standard ◽  
Lysosomal Storage Diseases ◽  
Krabbe Disease ◽  
Tandem Mass ◽  
Lysosomal Storage ◽  
Mass Detection ◽  
Storage Diseases

Abstract Background Deficiency of the lysosomal enzyme galactosylcerebrosidase (GALC) causes Krabbe disease. The diagnosis for Krabbe disease includes measurement of GALC enzymatic activity by radioisotope assay or accumulation of metabolite psychosine. To improve current diagnostic workflow and assay performance, we developed and validated a leukocyte enzymatic assay by using liquid chromatography tandem mass spectrometry (LC-MS/MS) for lysosomal storage diseases. Methods Leukocytes were separated and extracted from whole blood samples, and total protein was quantitated by BCA method. Commercialized and multiplexed substrates, internal standards, and buffer were incubated with cell lysates. The lysosomal enzymes in leukocytes metabolized the artificial substrate into product which is structurally identical to the internal standard. Liquid-liquid extraction was performed and supernatant was dried down and reconstituted. Liquid chromatography separation was achieved by Waters CSH C18, 2.1 x 50 mm column and Acquity UPLC system. A Waters Xevo TQS tandem mass spectrometer was used for mass detection. Results Enzymatic reaction products for six lysosomal enzymes were chromatographically resolved from substrate breakdown products through 3.5 minutes gradient liquid chromatography. Intra-assay imprecision was determined by 11 replicates of samples containing low and high concentration (CV<15%). Carryover was determined by assaying triplicates of cell lysate-free cocktails directly after injection of high enzyme activity sample (less than 0.1%). No matrix effect was found. The GALC enzyme activity was calculated and standardized by corresponding product and internal standard ratios from 5-point standard curve. The range of enzyme activity from three, known affected patients is 0.01–0.07 (nmol/hr/mg protein); whereas, two identified carriers had enzyme activate in the range of 0.14–0.40 (nmol/hr/mg protein). The reference interval was established from 63 residual, unaffected samples and was 0.12–5.97 (1.44±1.44) nmol/hr/mg protein. Conclusions A simple and multiplexed LC-MS/MS assay was developed which can measure small amounts of residual GALC enzyme activity in leukocytes. This confirmatory assay will aid in the diagnosis and prognosis (i.e. differentiate disease severity) of Krabbe disease and other lysosomal storage disorders.

scholarly journals Development of Organelle Replacement Therapy Using a Stearyl-Polyhistidine Peptide against Lysosomal Storage Disease Cells

Molecules ◽  
2019 ◽  
Vol 24 (16) ◽  
pp. 2995 ◽  
Author(s):  
Hayashi ◽  
Okamoto ◽  
Kawano ◽  
Iwasaki
Keyword(s):  
Replacement Therapy ◽  
Lysosomal Storage Disease ◽  
Lysosomal Enzymes ◽  
Drug Delivery Systems ◽  
Metabolic Disorders ◽  
Storage Disease ◽  
Cell Penetrating Peptide ◽  
Lysosomal Storage ◽  
Hydrophobic Region ◽  
Normal Cells

We previously reported on a polyhistidine peptide, His16 peptide, as a new cell-penetrating peptide. This peptide is anticipated to be a new carrier for drug delivery systems (DDSs) for targeting intracellular lysosomes because it can transport macromolecules (e.g., liposomes) into these organelles. In the present study, we examined the application of His16 peptide as a DDS carrier against lysosomal storage disease (LSD) cells. LSDs are metabolic disorders caused by loss of specific lysosomal enzymes. For the treatment of LSD cells, we devised a system designated organelle replacement therapy (ORT). ORT is a strategy for transporting exogenous lysosomes containing all kinds of lysosomal enzymes from normal cells into endogenous lysosomes in LSD cells using His16 peptide. To develop the ORT system, we prepared His16 peptide-modified healthy lysosomes (His16-Lyso) by insertion of a stearyl-His16 peptide into a hydrophobic region in the lysosomal membrane. His16-Lyso showed cellular uptake and localization to endogenous lysosomes in LSD cells. His16-Lyso also restored the proliferation of LSD cells, which otherwise showed slower proliferation than normal cells. These results suggested that His16-Lyso replenished deficient lysosomal enzymes in LSD cells. The results further suggest that His16-Lyso are promising candidates as a treatment tool for LSD cells and to establish a foundation for ORT.

scholarly journals Transcriptomic Changes Related to Cellular Processes with Particular Emphasis on Cell Activation in Lysosomal Storage Diseases from the Group of Mucopolysaccharidoses

2020 ◽  
Vol 21 (9) ◽  
pp. 3194 ◽  
Author(s):  
Estera Rintz ◽  
Lidia Gaffke ◽  
Magdalena Podlacha ◽  
Joanna Brokowska ◽  
Zuzanna Cyske ◽  
...  
Keyword(s):  
Cell Activation ◽  
Molecular Mechanisms ◽  
Metabolic Diseases ◽  
Lysosomal Storage Diseases ◽  
Lysosomal Storage ◽  
Storage Diseases ◽  
Monogenic Disorders ◽  
Cellular Processes ◽  
Transcriptomic Changes ◽  
Molecular Pathomechanisms

Although mucopolysaccharidoses (MPS), inherited metabolic diseases from the group of lysosomal storage diseases (LSD), are monogenic disorders, recent studies indicated that their molecular mechanisms are complicated. Storage of glycosaminoglycans (GAGs), arising from a deficiency in one of the enzymes involved in the degradation of these compounds, is the primary cause of each MPS type. However, dysfunctions of various cellular organelles and disturbance of cellular processes have been reported which contribute considerably to pathomechanisms of the disease. Here, we present a complex transcriptomic analysis in which all types and subtypes of MPS were investigated, with special emphasis on genes related to cell activation processes. Complex changes in expression of these genes were found in fibroblasts of all MPS types, with number of transcripts revealing higher or lower levels (relative to control fibroblasts) between 19 and over 50, depending on MPS type. Genes in which expression was significantly affected in most MPS types code for proteins involved in following processes, classified according to Gene Ontology knowledge database: cell activation, cell growth, cell recognition, and cell division. Levels of some transcripts (including CD9, CLU, MME and others) were especially significantly changed (over five times relative to controls). Our results are discussed in the light of molecular pathomechanisms of MPS, indicating that secondary and/or tertiary changes, relative to GAG storage, might significantly modulate cellular dysfunctions and contribute to molecular mechanisms of the disease. This may influence the efficacy of various therapies and suggests why various treatments are not fully effective in improving the complex symptoms of MPS.

scholarly journals Tandem Mass Spectrometry Has a Larger Analytical Range than Fluorescence Assays of Lysosomal Enzymes: Application to Newborn Screening and Diagnosis of Mucopolysaccharidoses Types II, IVA, and VI

2015 ◽  
Vol 61 (11) ◽  
pp. 1363-1371 ◽  
Author(s):  
Arun Babu Kumar ◽  
Sophia Masi ◽  
Farideh Ghomashchi ◽  
Naveen Kumar Chennamaneni ◽  
Makoto Ito ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Newborn Screening ◽  
Lysosomal Enzymes ◽  
Lysosomal Storage Diseases ◽  
Tandem Mass ◽  
Lysosomal Storage ◽  
Storage Diseases ◽  
Mps Ii ◽  
Analytical Range ◽  
Screening And Diagnosis

Abstract BACKGROUND There is interest in newborn screening and diagnosis of lysosomal storage diseases because of the development of treatment options that improve clinical outcome. Assays of lysosomal enzymes with high analytical range (ratio of assay response from the enzymatic reaction divided by the assay response due to nonenzymatic processes) are desirable because they are predicted to lead to a lower rate of false positives in population screening and to more accurate diagnoses. METHODS We designed new tandem mass spectrometry (MS/MS) assays that give the largest analytical ranges reported to date for the use of dried blood spots (DBS) for detection of mucopolysaccharidoses type II (MPS-II), MPS-IVA, and MPS-VI. For comparison, we carried out fluorometric assays of 6 lysosomal enzymes using 4-methylumbelliferyl (4MU)-substrate conjugates. RESULTS The MS/MS assays for MPS-II, -IVA, and -VI displayed analytical ranges that are 1–2 orders of magnitude higher than those for the corresponding fluorometric assays. The relatively small analytical ranges of the 4MU assays are due to the intrinsic fluorescence of the 4MU substrates, which cause high background in the assay response. CONCLUSIONS These highly reproducible MS/MS assays for MPS-II, -IVA, and -VI can support multiplex newborn screening of these lysosomal storage diseases. MS/MS assays of lysosomal enzymes outperform 4MU fluorometric assays in terms of analytical range. Ongoing pilot studies will allow us to gauge the impact of the increased analytical range on newborn screening performance.

scholarly journals Ferroptosis and Its Modulation by Autophagy in Light of the Pathogenesis of Lysosomal Storage Diseases

Cells ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 365
Author(s):  
Karolina Pierzynowska ◽  
Estera Rintz ◽  
Lidia Gaffke ◽  
Grzegorz Węgrzyn
Keyword(s):  
Oxidative Stress ◽  
Molecular Mechanisms ◽  
Lysosomal Storage Diseases ◽  
Review Article ◽  
Lysosomal Storage ◽  
Ferrous Ions ◽  
Autophagy Flux ◽  
Considerable Contribution ◽  
Storage Diseases

Ferroptosis is one of the recently described types of cell death which is dependent on many factors, including the accumulation of iron and lipid peroxidation. Its induction requires various signaling pathways. Recent discovery of ferroptosis induction pathways stimulated by autophagy, so called autophagy-dependent ferroptosis, put our attention on the role of ferroptosis in lysosomal storage diseases (LSD). Lysosome dysfunction, observed in these diseases, may influence ferroptosis efficiency, with as yet unknown consequences for the function of cells, tissues, and organisms, due to the effects of ferroptosis on physiological and pathological metabolic processes. Modulation of levels of ferrous ions and enhanced oxidative stress, which are primary markers of ferroptosis, are often described as processes associated with the pathology of LSD. Inhibition of autophagy flux and resultant accumulation of autophagosomes in neuronopathic LSD may induce autophagy-dependent ferroptosis, indicating a considerable contribution of this process in neurodegeneration. In this review article, we describe molecular mechanisms of ferroptosis in light of LSD, underlining the modulation of levels of ferroptosis markers in these diseases. Furthermore, we propose a hypothesis about the possible involvement of autophagy-dependent ferroptosis in these disorders.

scholarly journals Busulfan disposition below the age of three: alteration in children with lysosomal storage disease

Blood ◽  
1993 ◽  
Vol 82 (3) ◽  
pp. 1030-1034 ◽  
Author(s):  
G Vassal ◽  
A Fischer ◽  
D Challine ◽  
I Boland ◽  
F Ledheist ◽  
...  
Keyword(s):  
Young Children ◽  
Lysosomal Storage Disease ◽  
Storage Disease ◽  
Dose Range ◽  
Lysosomal Storage Diseases ◽  
Volume Of Distribution ◽  
Optimal Dosage ◽  
Lysosomal Storage ◽  
Acute Leukemias ◽  
Storage Diseases

Busulfan disposition is age-dependent with a higher clearance and a larger volume of distribution in children than in adults. The optimal dosage of busulfan needed to achieve bone marrow (BM) displacement in young children with malignant or nonmalignant disease remains to be defined. Using a gas chromatography-mass spectrometry assay, we evaluated plasma pharmacokinetics of busulfan in 33 children (median age, 9 months; range, 2 months to 2.75 years) with immune deficiencies, lysosomal storage diseases, acute leukemias, and malignant lymphohistiocytosis after an oral dose ranging from 0.9 to 2.6 mg/kg. The busulfan clearance (assuming a bioavailability of 1) ranged from 2.1 to 13.4 mL/min/kg with a mean of 6.8 mL/min/kg, which is higher than that reported in older children (4.5 mL/min/kg) and adults (2.9 mL/min/kg). Six children with lysosomal storage disease (5 with Hurler's disease, 1 with San Filippo's disease) had a prolonged elimination half-life (4.9 v 2.4 hours), a larger volume of distribution (3.4 v 1.2 L/kg) and a faster clearance (8.7 v 6.3 mL/min/kg) than the other 27 children. This suggests that a higher dose of busulfan will be required to achieve BM displacement in children with lysosomal storage disease. Over the dose range of 0.9 to 2.6 mg/kg, busulfan pharmacokinetics were linear. However, only 46% of the interpatient variation in systemic exposure could be ascribed to the dose. Given the wide interpatient variability in busulfan disposition, dose adjustment and drug monitoring will be needed to achieve the optimal dosage of busulfan in young children. The plasma busulfan levels required to achieve BM displacement need to be defined, especially in lysosomal storage diseases.

scholarly journals A Fourteen Years Old Boy with Cholesterol Ester Storage Disease

1970 ◽  
Vol 10 (2) ◽  
pp. 146-148
Author(s):  
Faizul Islum Chowdhury ◽  
Ahmedul Kabir ◽  
Jayanta Banik ◽  
Pinaki Paul ◽  
Mostofa Kamal ◽  
...  
Keyword(s):  
Liver Biopsy ◽  
Cholesterol Ester ◽  
Autosomal Recessive ◽  
Storage Disease ◽  
Specific Treatment ◽  
Autosomal Recessive Disorder ◽  
Lysosomal Storage Diseases ◽  
Lysosomal Storage ◽  
Acid Lipase ◽  
Storage Diseases

Cholesterol ester storage disease (CESD) is a rare autosomal recessive disorder resulting from lysosomal acid lipase deficiency and is usually characterized by hepatomegaly and hyperlipidemia. It is diagnosed by liver biopsy which characteristically shows microvesicular steatosis and periportal fibrosis. Here we report a fourteen years old boy who had presented with unexplained hepatomegaly, and hyperlipidemia determined incidentally. He was finally diagnosed as a case of cholesterol ester storage disease by liver biopsy. Though there is yet no specific treatment for CESD; however, the early detection of cases would make the timely control of complications possible. Keyword: Cholesterol Ester Storage Disease, Lysosomal Storage Diseases, Lipidoses    doi: 10.3329/jom.v10i2.2835   J MEDICINE 2009; 10 : 146-148

scholarly journals Busulfan disposition below the age of three: alteration in children with lysosomal storage disease

Blood ◽  
1993 ◽  
Vol 82 (3) ◽  
pp. 1030-1034 ◽  
Author(s):  
G Vassal ◽  
A Fischer ◽  
D Challine ◽  
I Boland ◽  
F Ledheist ◽  
...  
Keyword(s):  
Young Children ◽  
Lysosomal Storage Disease ◽  
Storage Disease ◽  
Dose Range ◽  
Lysosomal Storage Diseases ◽  
Volume Of Distribution ◽  
Optimal Dosage ◽  
Lysosomal Storage ◽  
Acute Leukemias ◽  
Storage Diseases

Abstract Busulfan disposition is age-dependent with a higher clearance and a larger volume of distribution in children than in adults. The optimal dosage of busulfan needed to achieve bone marrow (BM) displacement in young children with malignant or nonmalignant disease remains to be defined. Using a gas chromatography-mass spectrometry assay, we evaluated plasma pharmacokinetics of busulfan in 33 children (median age, 9 months; range, 2 months to 2.75 years) with immune deficiencies, lysosomal storage diseases, acute leukemias, and malignant lymphohistiocytosis after an oral dose ranging from 0.9 to 2.6 mg/kg. The busulfan clearance (assuming a bioavailability of 1) ranged from 2.1 to 13.4 mL/min/kg with a mean of 6.8 mL/min/kg, which is higher than that reported in older children (4.5 mL/min/kg) and adults (2.9 mL/min/kg). Six children with lysosomal storage disease (5 with Hurler's disease, 1 with San Filippo's disease) had a prolonged elimination half-life (4.9 v 2.4 hours), a larger volume of distribution (3.4 v 1.2 L/kg) and a faster clearance (8.7 v 6.3 mL/min/kg) than the other 27 children. This suggests that a higher dose of busulfan will be required to achieve BM displacement in children with lysosomal storage disease. Over the dose range of 0.9 to 2.6 mg/kg, busulfan pharmacokinetics were linear. However, only 46% of the interpatient variation in systemic exposure could be ascribed to the dose. Given the wide interpatient variability in busulfan disposition, dose adjustment and drug monitoring will be needed to achieve the optimal dosage of busulfan in young children. The plasma busulfan levels required to achieve BM displacement need to be defined, especially in lysosomal storage diseases.

Sign in / Sign up

Export Citation Format

Share Document