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Steven M. Levine - One of the best experts on this subject based on the ideXlab platform.
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substrate reduction intervention by l cycloserine in twitcher mice Globoid Cell Leukodystrophy on a b6 cast ei background
Neuroscience Letters, 2003Co-Authors: Sangita Biswas, Homigol Biesiada, Todd D Williams, Steven M. LevineAbstract:Abstract Globoid Cell Leukodystrophy (GCL) is usually a fatal demyelinating disease caused by mutations in galactosylceramidase, which normally recycles galactosylceramide, a predominant glycolipid of myelin, and psychosine. The initial pathology is thought to be due to the accumulation of psychosine in myelin-forming Cells leading to their death. In this study, substrate reduction therapy using l -cycloserine, an inhibitor of 3-ketodihydrosphingosine synthase, was examined in twitcher mice on a C57BL/6×CAST/Ei (B6;CAST/Ei) background, which mimics a late onset variant of GCL. A graded dose regimen of l -cycloserine initiated before the onset of symptoms increased the lifespan by ∼45% and delayed the onset of weight loss while the administration of l -cycloserine beginning after the onset of symptoms had no effect. Despite the pronounced effect for the early treatment regimen, B6;CAST/Ei twitcher mice still displayed a progressive disease leading to an early death.
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sphingolipid profile in the cns of the twitcher Globoid Cell Leukodystrophy mouse a lipidomics approach
Cellular and Molecular Biology, 2003Co-Authors: S W Esch, Sangita Biswas, T D Williams, Anuradha Chakrabarty, Steven M. LevineAbstract:Globoid Cell Leukodystrophy (Krabbe disease) is caused by mutations in galactosylceramidase, a lysosomal enzyme that acts to digest galactosylceramide, a glycolipid concentrated in myelin, and psychosine (galactosylsphingosine). Globoid Cell Leukodystrophy has been identified in many species including humans and twitcher mice. Several studies on human tissue have examined the lipid profile in this disease by gas, liquid or thin layer chromatography. Electrospray ionization tandem mass spectrometry combined with reverse phase HPLC has become a powerful alternative strategy, used here to compare the sphingolipid profile of pons/medulla tissue from twitcher mice with control tissue. In this lipidomics LC-MS approach, we scanned for precursors of m/z 264 to obtain a semi-quantitative profile of ceramides and galactosylceramides. Sphingosine-1-phosphate, C18:0 ceramide, C22:0 ceramide and C24:0 ceramide levels were reduced in the pons/medulla of twitcher mice compared to levels in control mice at 31 and 35-37 days of age. The levels of C22:0 and C24:0 galactosylceramide were similar between twitcher and control specimens and there was a trend toward reduced levels of C24:1 galactosylceramide and C24:1 hydroxy-galactosylceramide in twitcher specimens. Psychosine, C 16:0 ceramide and C 18:0 galactosylceramide levels were increased in the CNS of twitcher mice compared to levels in control mice. These data indicate that there is a trend toward decreased levels of long chain fatty acids and increased levels of shorter chain fatty acids in galactosylceramides and ceramides from twitcher mice compared with control mice, and such changes may be due to demyelination characteristic of acute pathology.
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delayed clinical and pathological signs in twitcher Globoid Cell Leukodystrophy mice on a c57bl 6 cast ei background
Neurobiology of Disease, 2002Co-Authors: Sangita Biswas, Homigol Biesiada, Todd D Williams, Steven M. LevineAbstract:Abstract Modifier genes may account for the phenotypic variability observed in the late-onset forms of Globoid Cell Leukodystrophy (GCL) in humans. In order to begin a search for modifier genes, the effect of genetic background on the clinical and pathological manifestations of GCL was investigated in twitcher mice. Twitcher mice on a C57BL/6 × CAST/Ei background had an increased life span (61.4 ± 2.5 vs 37.0 ± 0.6 days), a delayed onset of tremor (24 vs 21 days), and a delayed decline in walking ability compared to C57BL/6 twitcher mice. Pathologically, C57BL/6 × CAST/Ei twitcher mice had fewer lectin-positive Globoid Cells, less gliosis, and a greater preservation of myelin compared to C57BL/6 twitcher mice under moribund conditions. Similar concentrations of psychosine, the toxic species that accumulates in GCL, were measured by tandem mass spectrometry between moribund C57BL/6 twitcher mice (286.5 pmol/mg protein), 40-day C57BL/6 × CAST/Ei twitcher mice (276.5 pmol/mg), and moribund C57BL/6 × CAST/Ei twitcher mice (247.0 pmol/mg), suggesting that the milder phenotype in CAST/Ei × C57BL/6 twitcher mice did not correlate with less psychosine. In summary, the introduction of modifier genes from the wild, inbred CAST/Ei strain had a phenotypic effect resulting in a significantly slower disease course.
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substrate reduction therapy enhances the benefits of bone marrow transplantation in young mice with Globoid Cell Leukodystrophy
Pediatric Research, 2002Co-Authors: Sangita Biswas, Steven M. LevineAbstract:Globoid Cell Leukodystrophy is an autosomal recessive disease with progressive demyelination caused by a deficiency of the lysosomal enzyme galactosylceramidase. Bone marrow transplantation (BMT) is a therapeutic option for patients with late-onset disease and for patients with early onset disease that had an early diagnosis owing to an affected sibling. This therapy, however, typically is not effective for early onset disease when the diagnosis occurs after several months of life. In an effort to enable a broader range of patients to benefit from BMT, we tested whether combining substrate-reduction therapy with BMT would result in a greater benefit than either treatment alone in the twitcher mouse model of Globoid Cell Leukodystrophy. Twitcher mice treated with l-cycloserine, an inhibitor of 3-ketodyhydrosphingosine synthase, and transplanted with 50 ± 5 × 106 bone marrow Cells on d 10 had a mean life-span of 112 d compared with 51 d for BMT alone (p < 0.001) or l-cycloserine alone, which was previously reported to be 56 d. l-Cycloserine treatment also was initiated neonatally to determine whether it would allow for a delayed BMT to have therapeutic value. Twitcher mice given only BMT at 18 d or only a short course of l-cycloserine died at 36 and 37 d, respectively. Twitcher mice given a short course of l-cycloserine + BMT at 18 d lived to 58 d (p = 0.0006). In conclusion, substrate-reduction therapy enhanced the value of BMT in twitcher mice, suggesting that this combination strategy might benefit patients with Globoid Cell Leukodystrophy.
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il 6 deficiency allows for enhanced therapeutic value after bone marrow transplantation across a minor histocompatibility barrier in the twitcher Globoid Cell Leukodystrophy mouse
Journal of Neuroscience Research, 2001Co-Authors: Sangita Biswas, Illya G. Bronshteyn, David M. Pinson, Steven M. LevineAbstract:Bone marrow transplantation (BMT) has therapeutic value for twitcher (Globoid Cell Leukodystrophy) mice, which suffer from a genetic deficiency of the lysosomal enzyme galactosylceramidase that leads to progressive demyelination and early death. Preliminary investigations indicated that a semiallogeneic BMT resulted in graft vs. host disease (GVHD) in twitcher mice but not normal mice. Increased production of the cytokine IL-6 has been demonstrated in twitcher mice, and it has been linked with induction of GVHD. We investigated the effects of BMT in twitcher/IL-6 deficient mice and compared these findings with those from transplanted twitcher and control mice. After a semiallogeneic BMT, 11.4% of controls died within few weeks while the rest survived >100 days without GVHD. In contrast, 85% of the transplanted twitcher mice died by 70 days and 65% developed clinical signs of GVHD, e.g., alopecia and weight loss. In transplanted twitcher/IL-6 deficient mice, only 21% died by Day 70, none had alopecia, and 23% had weight loss. There was no difference in the onset day and severity of twitching between twitcher and twitcher/IL-6 deficient mice after BMT. In transplanted twitcher/IL-6 deficient mice, there was improvement of BBB integrity and a decrease in Globoid Cell number compared with nontransplanted twitcher/IL-6 deficient mice. In summary, these results demonstrate that an underlying pathology like Globoid Cell Leukodystrophy leads to activation of GVHD responses in a donor–host combination that would not normally induce GVHD. Furthermore, IL-6 seems to play a key role because a deficiency of IL-6 results in a better prognosis. J. Neurosci. Res. 65:298–307, 2001. © 2001 Wiley-Liss, Inc.
D. A. Wenger - One of the best experts on this subject based on the ideXlab platform.
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enzyme replacement therapy of a novel humanized mouse model of Globoid Cell Leukodystrophy
Experimental Neurology, 2015Co-Authors: Frank Matthes, D. A. Wenger, Claes Andersson, Axel Stein, Carl Eistrup, Jens Fogh, Volkmar Gieselmann, Ulrich MatznerAbstract:An inherited deficiency of β-galactosylceramidase (GALC) causes the lysosomal storage disease Globoid Cell Leukodystrophy (GLD). The disease is characterized by the accumulation of the cytotoxic metabolite psychosine (galactosylsphingosine), causing rapid degeneration of myelinating Cells. Most patients suffer from the infantile form of GLD with onset of disease between 3 and 6 months after birth and death by 2 years of age. The most widely used animal model of GLD, the twitcher mouse, presents with an even more rapid course of disease and death around 40 days of age. We have generated a novel "humanized" mouse model of GLD by inserting a human GALC cDNA containing an adult-onset patient mutation into the murine GALC gene. Humanized GALC mice exhibit pathological hallmarks of GLD including psychosine accumulation, neuroinflammation, CNS infiltration of macrophages, astrogliosis and demyelination. Residual GALC activities in mouse tissues are low and the mice display a median lifespan of 46 days. Due to the expression of the human transgene, the mice do not develop an immune response against rhGALC, rendering the animal model suitable for therapies based on human enzyme. Intravenously injected rhGALC was able to surmount the blood-brain barrier and was targeted to lysosomes of brain macrophages, astrocytes and neurons. High-dose enzyme replacement therapy started at postnatal day 21 reduced the elevated psychosine levels in the peripheral and central nervous system by 14-16%, but did not ameliorate neuroinflammation, demyelination and lifespan. These results may indicate that treatment must be started earlier before pathology occurs.
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krabbe disease Globoid Cell Leukodystrophy
2015Co-Authors: D. A. Wenger, Paola LuziAbstract:Abstract Krabbe disease or Globoid Cell Leukodystrophy (GLD) is an autosomal recessive disorder of myelination caused by galactocerebrosidase (GALC) deficiency. This enzyme is essential for the lysosomal degradation of several important galactolipids, including galactosylceramide and psychosine. Most human patients present before 6 months of age but older patients are also recognized. The diagnosis can be made by measuring very low GALC activity in any easily obtainable tissue. Newborn screening has been initiated in an attempt to identify individuals who may develop the disease before symptoms appear. Treatment is limited to hematopoietic stem Cell transplantation in presymptomatic infants and mildly affected late-onset patients. The GALC gene has been cloned, and over 140 disease-causing mutations have been identified. GLD has also been identified in a number of animal species, which can be used to examine pathogenic mechanisms and potential methods for treatment including gene therapy, stem Cell therapy, and small molecule therapy.
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chapter 30 krabbe disease Globoid Cell Leukodystrophy
Rosenberg's Molecular and Genetic Basis of Neurological and Psychiatric Disease (Fifth Edition), 2015Co-Authors: D. A. Wenger, Paola LuziAbstract:Krabbe disease or Globoid Cell Leukodystrophy (GLD) is an autosomal recessive disorder of myelination caused by galactocerebrosidase (GALC) deficiency. This enzyme is essential for the lysosomal degradation of several important galactolipids, including galactosylceramide and psychosine. Most human patients present before 6 months of age but older patients are also recognized. The diagnosis can be made by measuring very low GALC activity in any easily obtainable tissue. Newborn screening has been initiated in an attempt to identify individuals who may develop the disease before symptoms appear. Treatment is limited to hematopoietic stem Cell transplantation in presymptomatic infants and mildly affected late-onset patients. The GALC gene has been cloned, and over 140 disease-causing mutations have been identified. GLD has also been identified in a number of animal species, which can be used to examine pathogenic mechanisms and potential methods for treatment including gene therapy, stem Cell therapy, and small molecule therapy.
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effects of treatments on inflammatory and apoptotic markers in the cns of mice with Globoid Cell Leukodystrophy
Brain Research, 2009Co-Authors: Paola Luzi, Mohammad A Rafi, Mark T Curtis, Ronnie M Abraham, Craig D Hooper, D. A. WengerAbstract:Globoid Cell Leukodystrophy (GLD) or Krabbe disease is a neurodegenerative disorder caused by the deficiency of the lysosomal enzyme galactocerebrosidase (GALC). GALC deficiency results in a progressive demyelination of the central and peripheral nervous systems. Inflammatory Cells and increased levels of cytokines and chemokines are present in the CNS of GLD mice and may play a significant role in the pathogenesis of the disease. In this study we evaluate the effect of non-steroidal anti-inflammatory drugs, such as indomethacin and ibuprofen, and minocycline, a tetracycline analog with neuroprotective and anti-apoptotic properties, on the progression of the disease using a transgenic mouse model of GLD. Real-time quantitative PCR was used to analyze the expression of several markers of the immune/inflammatory response. IL-6, TNF-α, MIP-1β, MCP-1, iNOS/NOS2, CD11b, CD68, CD4 and CD8 mRNA levels were measured in cortex, cerebellum and spinal cord of untreated and treated affected mice at different ages. In addition, the pharmacological treatments were compared to bone marrow transplantation (BMT). The pharmacological treatments significantly extended the life-span of the treated mice and reduced the levels of several of the immuno-related factors studied. However, BMT produced the most dramatic improvements. In BMT-treated mice, factors in the spinal cord were normalized faster than the cerebellum, with the exception of CD68. There was a decrease in the number of apoptotic Cells in the cerebellum of mice receiving anti-inflammatory drugs and BMT. These studies indicate a possible role for combined therapy in the treatment of GLD.
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aav2 5 vector expressing galactocerebrosidase ameliorates cns disease in the murine model of Globoid Cell Leukodystrophy more efficiently than aav2
Molecular Therapy, 2005Co-Authors: Darshong Lin, D. A. Wenger, Carole Vogler, Corinne R Fantz, Beth Levy, Mohammad A Rafi, Mark S SandsAbstract:Globoid-Cell Leukodystrophy (GLD) is an autosomal recessive lysosomal storage disorder caused by mutations in the galactosylceramidase (GALC) gene. Infantile GLD has a lethal course with severe cerebral demyelination that progresses to death by 2 years of age. In the current study twitcher mice, an authentic murine model of infantile GLD, were given intracranial injections of either recombinant adeno-associated virus serotype 2 encoding the murine Galc cDNA (AAV2-GALC) or the same genome pseudotyped with AAV5 capsid proteins (AAV2/5-GALC) on day 3 of age. The group injected intracranially with AAV2/5-GALC had approximately 25-fold greater than normal Galc levels in the brain, while AAV2-GALC-injected animals had 28% normal levels. The average life expectancy of twitcher mice ( approximately 38 days) was significantly (P < 0.0001) increased to 48 and 52 days for the AAV2-GALC- and AAV2/5-GALC-treated groups, respectively. The AAV2/5-GALC group performed significantly better in a battery of behavioral tests compared to untreated, AAV2-GFP-treated, or AAV2-treated twitcher animals. This longitudinal study demonstrated that AAV2/5-GALC-mediated gene therapy resulted in higher levels of Galc expression and slowed the neurologic deterioration more completely than AAV2-GALC in the murine model of Globoid-Cell Leukodystrophy. However, the clinical improvements, as assessed by behavioral tests and life span, were only modest.
William Krivit - One of the best experts on this subject based on the ideXlab platform.
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Globoid Cell Leukodystrophy krabbe disease normal umbilical cord blood galactocerebrosidase activity and polymorphic mutations
Journal of Inherited Metabolic Disease, 2005Co-Authors: S S Raghavan, Joanne Kurtzberg, Edwin H. Kolodny, B J Zeng, Paola Torres, Gregory M Pastores, William KrivitAbstract:Globoid Cell Leukodystrophy is an inherited metabolic disorder of the central nervous system caused by deficiency of the lysosomal enzyme galactocerebrosidase. Haematopoietic stem Cell transplantation is the only available effective treatment. The engraftment from normal donors provides competent Cells able to correct the metabolic defect. Umbilical cord blood Cells have proved to significantly decrease complications and improve engraftment rate compared to adult marrow Cells in haematopoietic stem Cell transplantation. Umbilical cord blood Cells must be of sufficient activity to provide central nervous system recovery after engraftment is obtained. Galactocerebrosidase activity is known to be affected by two polymorphic alleles found at nucleotides 502 and 1637 of the cDNA for this gene. This enzyme activity and the polymorphic alleles noted above were analysed in 83 random samples of umbilical cord blood. The activity, assayed with the fluorogenic substrate 6-hexadecanoylamino-4-methylumbelliferyl-beta-galactopyranoside, in those with neither polymorphic allele was 4.6 +/- 1.7 units (nmol/h per mg protein). This optimal choice of cord blood was found in only 24% of specimens. Homozygotes for 1637T > C with activity of only 1.5 +/- 0.4 units represented 16% of the samples. Those heterozygous for 1637T > C with slightly better activity (2.3 +/- 0.7 units) represented 52% of the samples. Choice of umbilical cord blood for haematopoietic stem Cell transplantation, therefore, requires consideration not only of Cell quantity and HLA compatibility but also selection for normal alleles to obtain maximal enzymatic activity for central nervous system correction.
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measurements from normal umbilical cord blood of four lysosomal enzymatic activities α l iduronidase hurler galactocerebrosidase Globoid Cell Leukodystrophy arylsulfatase a metachromatic Leukodystrophy arylsulfatase b maroteaux lamy
Bone Marrow Transplantation, 2000Co-Authors: Rita Degasperi, S S Raghavan, C Carrier, P Rubenstein, Magama Sosa, Joanne Kurtzberg, Charles Peters, John E Wagner, Edwin H. Kolodny, William KrivitAbstract:Measurements from normal umbilical cord blood of four lysosomal enzymatic activities: α- L -iduronidase (Hurler), galactocerebrosidase (Globoid Cell Leukodystrophy), arylsulfatase A (metachromatic Leukodystrophy), arylsulfatase B (Maroteaux-Lamy)
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Globoid Cell Leukodystrophy distinguishing early onset from late onset disease using a brain mr imaging scoring method
American Journal of Neuroradiology, 1999Co-Authors: Daniel J Loes, Charles Peters, William KrivitAbstract:BACKGROUND AND PURPOSE: Our purpose was to determine the characteristic MR features of early-onset (before age 2 years) versus late-onset (after age 2 years) Globoid Cell Leukodystrophy (GLD). METHODS: Thirty-four brain MR images in 22 patients with GLD were reviewed. A severity score (0 to 32), based on a point system derived from the location and extent of disease and the presence of focal and/or global atrophy, was calculated for each examination. RESULTS: Of the 22 patients, three were asymptomatic and 19 were symptomatic. Ten patients had early-onset disease, whereas nine had late-onset disease. MR images of all patients showed abnormalities. In the early-onset group (n = 10; mean maximum MR score, 8.1; range, 3–18), 90% had pyramidal tract involvement, 80% had cerebellar white matter involvement, 70% had deep gray matter involvement, 60% had posterior corpus callosal involvement, 50% had parietooccipital white matter involvement, and 40% had cerebral atrophy. Serial MR imaging in four of these patients revealed progressive disease. In the late-onset group (n = 9; mean maximum MR score, 5.6; range, 4–10), 100% had pyramidal tract involvement, 100% had parietooccipital white matter involvement, 89% had posterior corpus callosal involvement, and none had cerebellar white matter involvement, deep gray matter involvement, or cerebral atrophy. Serial MR imaging in one patient with late-onset GLD did not reveal any change. A spectrum of findings was observed in the three patients who were asymptomatic. CONCLUSION: Cerebellar white matter and deep gray matter involvement are present only in early-onset GLD. Pyramidal tract involvement is a characteristic finding in both early- and late-onset GLD. This scoring method for brain MR observations will assist in the objective assessment of the impact of hematopoietic stem Cell transplantation in patients with GLD.
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hematopoietic stem Cell transplantation in Globoid Cell Leukodystrophy
The New England Journal of Medicine, 1998Co-Authors: William Krivit, Joanne Kurtzberg, Charles Peters, John E Wagner, Edwin H. Kolodny, D. A. Wenger, Elsa Shapiro, Guy Cornu, Marie T Vanier, Daniel J LoesAbstract:BACKGROUND: Globoid-Cell Leukodystrophy is caused by a deficiency of galactocerebrosidase, which results in progressive central nervous system deterioration. We investigated whether allogeneic hematopoietic stem-Cell transplantation can provide a source of leukocyte galactocerebrosidase and thereby prevent the decline of central nervous system function in patients with the disease. METHODS: Five children with Globoid-Cell Leukodystrophy (one with the infantile type and four with late-onset disease) were treated with allogeneic hematopoietic stem-Cell transplantation. Measurement of leukocyte galactocerebrosidase levels, neurologic examinations, neuropsychological tests, magnetic resonance imaging of the central nervous system, cerebrospinal fluid protein assays, and neurophysiologic measurements were performed before and after transplantation, with follow-up ranging from one to nine years. RESULTS: Engraftment of donor-derived hematopoietic Cells occurred in all patients and was followed by restoration of normal leukocyte galactocerebrosidase levels. In the four patients with late-onset disease, the central nervous system deterioration was reversed, and in the patient with the infantile form of the disease, signs and symptoms have not appeared. Magnetic resonance imaging showed a decrease in signal intensity in the three patients with late-onset disease who were assessed both before and after transplantation. Abnormalities in cerebrospinal fluid total protein levels were corrected in three patients with late-onset disease and substantially reduced in the patient with the infantile form. CONCLUSIONS:Central nervous system manifestations of Globoid-Cell Leukodystrophy can be reversed by allogeneic hematopoietic stem-Cell transplantation.
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molecular heterogeneity of late onset forms of Globoid Cell Leukodystrophy
American Journal of Human Genetics, 1996Co-Authors: Rita De Gasperi, William Krivit, Edi Lucia Sartorato, Stefania Battistini, M Gama A Sosa, H Macfarlane, J F Gusella, Edwin H. KolodnyAbstract:Abstract Globoid-Cell Leukodystrophy (GLD) is an autosomal recessive inherited disorder caused by the deficiency of galactocerebrosidase, the lysosomal enzyme responsible for the degradation of the myelin glycolipid galactocerebroside. Although the most common form of the disease is the classical infantile form (Krabbe disease), later-onset forms also have been described. We have analyzed the galactocerebrosidase gene in 17 patients (nine families) with late-onset GLD and in 1 patient with classical Krabbe disease. Half of the patients were heterozygous for the large gene deletion associated with the 502C-->T polymorphism, the most common mutation in infantile patients. Several novel mutations that result in deficient galactocerebrosidase activity were also identified in these patients. They include the missense mutations R63H, G95S, M101L, G268S, Y298C, and I234T; the nonsense mutation S7X; a one-base deletion (805delG); a mutation that interferes with the splicing of intron 1; and a 34-nt insertion in the RNA, caused by the aberrant splicing of intron 6. All of these genetic defects are clustered in the first 10 exons of the galactocerebrosidase gene and therefore affect the 50-kD subunit of the mature enzyme. Studies on the distribution and enzymatic activity of the polymorphic alleles 1637T/C (I546/T546) provided support for previous data that had indicated the existence of two galactocerebrosidase forms with different catalytic activities in the general population. Our data also indicate that the mutations occur preferentially in the "low activity" 1637C allele.
Edwin H. Kolodny - One of the best experts on this subject based on the ideXlab platform.
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Globoid Cell Leukodystrophy krabbe disease normal umbilical cord blood galactocerebrosidase activity and polymorphic mutations
Journal of Inherited Metabolic Disease, 2005Co-Authors: S S Raghavan, Joanne Kurtzberg, Edwin H. Kolodny, B J Zeng, Paola Torres, Gregory M Pastores, William KrivitAbstract:Globoid Cell Leukodystrophy is an inherited metabolic disorder of the central nervous system caused by deficiency of the lysosomal enzyme galactocerebrosidase. Haematopoietic stem Cell transplantation is the only available effective treatment. The engraftment from normal donors provides competent Cells able to correct the metabolic defect. Umbilical cord blood Cells have proved to significantly decrease complications and improve engraftment rate compared to adult marrow Cells in haematopoietic stem Cell transplantation. Umbilical cord blood Cells must be of sufficient activity to provide central nervous system recovery after engraftment is obtained. Galactocerebrosidase activity is known to be affected by two polymorphic alleles found at nucleotides 502 and 1637 of the cDNA for this gene. This enzyme activity and the polymorphic alleles noted above were analysed in 83 random samples of umbilical cord blood. The activity, assayed with the fluorogenic substrate 6-hexadecanoylamino-4-methylumbelliferyl-beta-galactopyranoside, in those with neither polymorphic allele was 4.6 +/- 1.7 units (nmol/h per mg protein). This optimal choice of cord blood was found in only 24% of specimens. Homozygotes for 1637T > C with activity of only 1.5 +/- 0.4 units represented 16% of the samples. Those heterozygous for 1637T > C with slightly better activity (2.3 +/- 0.7 units) represented 52% of the samples. Choice of umbilical cord blood for haematopoietic stem Cell transplantation, therefore, requires consideration not only of Cell quantity and HLA compatibility but also selection for normal alleles to obtain maximal enzymatic activity for central nervous system correction.
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measurements from normal umbilical cord blood of four lysosomal enzymatic activities α l iduronidase hurler galactocerebrosidase Globoid Cell Leukodystrophy arylsulfatase a metachromatic Leukodystrophy arylsulfatase b maroteaux lamy
Bone Marrow Transplantation, 2000Co-Authors: Rita Degasperi, S S Raghavan, C Carrier, P Rubenstein, Magama Sosa, Joanne Kurtzberg, Charles Peters, John E Wagner, Edwin H. Kolodny, William KrivitAbstract:Measurements from normal umbilical cord blood of four lysosomal enzymatic activities: α- L -iduronidase (Hurler), galactocerebrosidase (Globoid Cell Leukodystrophy), arylsulfatase A (metachromatic Leukodystrophy), arylsulfatase B (Maroteaux-Lamy)
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molecular basis of late life Globoid Cell Leukodystrophy
Human Mutation, 1999Co-Authors: Rita De Gasperi, S S Raghavan, Miguel Gama A Sosa, Edi Lucia Sartorato, Stefania Battistini, Edwin H. KolodnyAbstract:Globoid Cell Leukodystrophy is an autosomal recessive inherited disease caused by deficiency of the lysosomal enzyme galactocerebrosidase (GALC). Although the severe, rapidly progressing infantile form is the most common, late-onset forms have been described. We investigated the molecular basis of GALC deficiency in a patient with a late-life mild form of Globoid Cell Leukodystrophy who survived into the eighth decade. Since material suitable for mutation analysis was no longer available from the proband, her GALC genotype was reconstructed by analyzing this gene in her six obligate carrier offspring. One allele contained the mutation 809G>A (G270D) in the 1637C background, while the other allele contained three sequence variants: 1609G>A (G537R), 1873G>A (A625T), and 1650T>A (V550V) in the 1637T background. These mutations were confirmed in the proband's genomic DNA isolated from a sural nerve biopsy. Expression studies indicated that the G537R is a disease-causing mutation, as it resulted in no GALC activity, either alone or together with the A625T. This A625T sequence variant did not affect the enzyme activity, at least when expressed in the 1637T background. The mild clinical phenotype was likely to be associated with the 809G>A, since residual GALC activity, about 17% of the control activity, was detected in the expression studies of this mutation. This mutation has been found in several other patients with late-onset GLD. Hum Mutat 14:256–262, 1999. © 1999 Wiley-Liss, Inc.
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hematopoietic stem Cell transplantation in Globoid Cell Leukodystrophy
The New England Journal of Medicine, 1998Co-Authors: William Krivit, Joanne Kurtzberg, Charles Peters, John E Wagner, Edwin H. Kolodny, D. A. Wenger, Elsa Shapiro, Guy Cornu, Marie T Vanier, Daniel J LoesAbstract:BACKGROUND: Globoid-Cell Leukodystrophy is caused by a deficiency of galactocerebrosidase, which results in progressive central nervous system deterioration. We investigated whether allogeneic hematopoietic stem-Cell transplantation can provide a source of leukocyte galactocerebrosidase and thereby prevent the decline of central nervous system function in patients with the disease. METHODS: Five children with Globoid-Cell Leukodystrophy (one with the infantile type and four with late-onset disease) were treated with allogeneic hematopoietic stem-Cell transplantation. Measurement of leukocyte galactocerebrosidase levels, neurologic examinations, neuropsychological tests, magnetic resonance imaging of the central nervous system, cerebrospinal fluid protein assays, and neurophysiologic measurements were performed before and after transplantation, with follow-up ranging from one to nine years. RESULTS: Engraftment of donor-derived hematopoietic Cells occurred in all patients and was followed by restoration of normal leukocyte galactocerebrosidase levels. In the four patients with late-onset disease, the central nervous system deterioration was reversed, and in the patient with the infantile form of the disease, signs and symptoms have not appeared. Magnetic resonance imaging showed a decrease in signal intensity in the three patients with late-onset disease who were assessed both before and after transplantation. Abnormalities in cerebrospinal fluid total protein levels were corrected in three patients with late-onset disease and substantially reduced in the patient with the infantile form. CONCLUSIONS:Central nervous system manifestations of Globoid-Cell Leukodystrophy can be reversed by allogeneic hematopoietic stem-Cell transplantation.
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molecular heterogeneity of late onset forms of Globoid Cell Leukodystrophy
American Journal of Human Genetics, 1996Co-Authors: Rita De Gasperi, William Krivit, Edi Lucia Sartorato, Stefania Battistini, M Gama A Sosa, H Macfarlane, J F Gusella, Edwin H. KolodnyAbstract:Abstract Globoid-Cell Leukodystrophy (GLD) is an autosomal recessive inherited disorder caused by the deficiency of galactocerebrosidase, the lysosomal enzyme responsible for the degradation of the myelin glycolipid galactocerebroside. Although the most common form of the disease is the classical infantile form (Krabbe disease), later-onset forms also have been described. We have analyzed the galactocerebrosidase gene in 17 patients (nine families) with late-onset GLD and in 1 patient with classical Krabbe disease. Half of the patients were heterozygous for the large gene deletion associated with the 502C-->T polymorphism, the most common mutation in infantile patients. Several novel mutations that result in deficient galactocerebrosidase activity were also identified in these patients. They include the missense mutations R63H, G95S, M101L, G268S, Y298C, and I234T; the nonsense mutation S7X; a one-base deletion (805delG); a mutation that interferes with the splicing of intron 1; and a 34-nt insertion in the RNA, caused by the aberrant splicing of intron 6. All of these genetic defects are clustered in the first 10 exons of the galactocerebrosidase gene and therefore affect the 50-kD subunit of the mature enzyme. Studies on the distribution and enzymatic activity of the polymorphic alleles 1637T/C (I546/T546) provided support for previous data that had indicated the existence of two galactocerebrosidase forms with different catalytic activities in the general population. Our data also indicate that the mutations occur preferentially in the "low activity" 1637C allele.
Sangita Biswas - One of the best experts on this subject based on the ideXlab platform.
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substrate reduction intervention by l cycloserine in twitcher mice Globoid Cell Leukodystrophy on a b6 cast ei background
Neuroscience Letters, 2003Co-Authors: Sangita Biswas, Homigol Biesiada, Todd D Williams, Steven M. LevineAbstract:Abstract Globoid Cell Leukodystrophy (GCL) is usually a fatal demyelinating disease caused by mutations in galactosylceramidase, which normally recycles galactosylceramide, a predominant glycolipid of myelin, and psychosine. The initial pathology is thought to be due to the accumulation of psychosine in myelin-forming Cells leading to their death. In this study, substrate reduction therapy using l -cycloserine, an inhibitor of 3-ketodihydrosphingosine synthase, was examined in twitcher mice on a C57BL/6×CAST/Ei (B6;CAST/Ei) background, which mimics a late onset variant of GCL. A graded dose regimen of l -cycloserine initiated before the onset of symptoms increased the lifespan by ∼45% and delayed the onset of weight loss while the administration of l -cycloserine beginning after the onset of symptoms had no effect. Despite the pronounced effect for the early treatment regimen, B6;CAST/Ei twitcher mice still displayed a progressive disease leading to an early death.
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sphingolipid profile in the cns of the twitcher Globoid Cell Leukodystrophy mouse a lipidomics approach
Cellular and Molecular Biology, 2003Co-Authors: S W Esch, Sangita Biswas, T D Williams, Anuradha Chakrabarty, Steven M. LevineAbstract:Globoid Cell Leukodystrophy (Krabbe disease) is caused by mutations in galactosylceramidase, a lysosomal enzyme that acts to digest galactosylceramide, a glycolipid concentrated in myelin, and psychosine (galactosylsphingosine). Globoid Cell Leukodystrophy has been identified in many species including humans and twitcher mice. Several studies on human tissue have examined the lipid profile in this disease by gas, liquid or thin layer chromatography. Electrospray ionization tandem mass spectrometry combined with reverse phase HPLC has become a powerful alternative strategy, used here to compare the sphingolipid profile of pons/medulla tissue from twitcher mice with control tissue. In this lipidomics LC-MS approach, we scanned for precursors of m/z 264 to obtain a semi-quantitative profile of ceramides and galactosylceramides. Sphingosine-1-phosphate, C18:0 ceramide, C22:0 ceramide and C24:0 ceramide levels were reduced in the pons/medulla of twitcher mice compared to levels in control mice at 31 and 35-37 days of age. The levels of C22:0 and C24:0 galactosylceramide were similar between twitcher and control specimens and there was a trend toward reduced levels of C24:1 galactosylceramide and C24:1 hydroxy-galactosylceramide in twitcher specimens. Psychosine, C 16:0 ceramide and C 18:0 galactosylceramide levels were increased in the CNS of twitcher mice compared to levels in control mice. These data indicate that there is a trend toward decreased levels of long chain fatty acids and increased levels of shorter chain fatty acids in galactosylceramides and ceramides from twitcher mice compared with control mice, and such changes may be due to demyelination characteristic of acute pathology.
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delayed clinical and pathological signs in twitcher Globoid Cell Leukodystrophy mice on a c57bl 6 cast ei background
Neurobiology of Disease, 2002Co-Authors: Sangita Biswas, Homigol Biesiada, Todd D Williams, Steven M. LevineAbstract:Abstract Modifier genes may account for the phenotypic variability observed in the late-onset forms of Globoid Cell Leukodystrophy (GCL) in humans. In order to begin a search for modifier genes, the effect of genetic background on the clinical and pathological manifestations of GCL was investigated in twitcher mice. Twitcher mice on a C57BL/6 × CAST/Ei background had an increased life span (61.4 ± 2.5 vs 37.0 ± 0.6 days), a delayed onset of tremor (24 vs 21 days), and a delayed decline in walking ability compared to C57BL/6 twitcher mice. Pathologically, C57BL/6 × CAST/Ei twitcher mice had fewer lectin-positive Globoid Cells, less gliosis, and a greater preservation of myelin compared to C57BL/6 twitcher mice under moribund conditions. Similar concentrations of psychosine, the toxic species that accumulates in GCL, were measured by tandem mass spectrometry between moribund C57BL/6 twitcher mice (286.5 pmol/mg protein), 40-day C57BL/6 × CAST/Ei twitcher mice (276.5 pmol/mg), and moribund C57BL/6 × CAST/Ei twitcher mice (247.0 pmol/mg), suggesting that the milder phenotype in CAST/Ei × C57BL/6 twitcher mice did not correlate with less psychosine. In summary, the introduction of modifier genes from the wild, inbred CAST/Ei strain had a phenotypic effect resulting in a significantly slower disease course.
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substrate reduction therapy enhances the benefits of bone marrow transplantation in young mice with Globoid Cell Leukodystrophy
Pediatric Research, 2002Co-Authors: Sangita Biswas, Steven M. LevineAbstract:Globoid Cell Leukodystrophy is an autosomal recessive disease with progressive demyelination caused by a deficiency of the lysosomal enzyme galactosylceramidase. Bone marrow transplantation (BMT) is a therapeutic option for patients with late-onset disease and for patients with early onset disease that had an early diagnosis owing to an affected sibling. This therapy, however, typically is not effective for early onset disease when the diagnosis occurs after several months of life. In an effort to enable a broader range of patients to benefit from BMT, we tested whether combining substrate-reduction therapy with BMT would result in a greater benefit than either treatment alone in the twitcher mouse model of Globoid Cell Leukodystrophy. Twitcher mice treated with l-cycloserine, an inhibitor of 3-ketodyhydrosphingosine synthase, and transplanted with 50 ± 5 × 106 bone marrow Cells on d 10 had a mean life-span of 112 d compared with 51 d for BMT alone (p < 0.001) or l-cycloserine alone, which was previously reported to be 56 d. l-Cycloserine treatment also was initiated neonatally to determine whether it would allow for a delayed BMT to have therapeutic value. Twitcher mice given only BMT at 18 d or only a short course of l-cycloserine died at 36 and 37 d, respectively. Twitcher mice given a short course of l-cycloserine + BMT at 18 d lived to 58 d (p = 0.0006). In conclusion, substrate-reduction therapy enhanced the value of BMT in twitcher mice, suggesting that this combination strategy might benefit patients with Globoid Cell Leukodystrophy.
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il 6 deficiency allows for enhanced therapeutic value after bone marrow transplantation across a minor histocompatibility barrier in the twitcher Globoid Cell Leukodystrophy mouse
Journal of Neuroscience Research, 2001Co-Authors: Sangita Biswas, Illya G. Bronshteyn, David M. Pinson, Steven M. LevineAbstract:Bone marrow transplantation (BMT) has therapeutic value for twitcher (Globoid Cell Leukodystrophy) mice, which suffer from a genetic deficiency of the lysosomal enzyme galactosylceramidase that leads to progressive demyelination and early death. Preliminary investigations indicated that a semiallogeneic BMT resulted in graft vs. host disease (GVHD) in twitcher mice but not normal mice. Increased production of the cytokine IL-6 has been demonstrated in twitcher mice, and it has been linked with induction of GVHD. We investigated the effects of BMT in twitcher/IL-6 deficient mice and compared these findings with those from transplanted twitcher and control mice. After a semiallogeneic BMT, 11.4% of controls died within few weeks while the rest survived >100 days without GVHD. In contrast, 85% of the transplanted twitcher mice died by 70 days and 65% developed clinical signs of GVHD, e.g., alopecia and weight loss. In transplanted twitcher/IL-6 deficient mice, only 21% died by Day 70, none had alopecia, and 23% had weight loss. There was no difference in the onset day and severity of twitching between twitcher and twitcher/IL-6 deficient mice after BMT. In transplanted twitcher/IL-6 deficient mice, there was improvement of BBB integrity and a decrease in Globoid Cell number compared with nontransplanted twitcher/IL-6 deficient mice. In summary, these results demonstrate that an underlying pathology like Globoid Cell Leukodystrophy leads to activation of GVHD responses in a donor–host combination that would not normally induce GVHD. Furthermore, IL-6 seems to play a key role because a deficiency of IL-6 results in a better prognosis. J. Neurosci. Res. 65:298–307, 2001. © 2001 Wiley-Liss, Inc.