Enzymatic characterization of novel arylsulfatase A variants using human arylsulfatase A-deficient immortalized mesenchymal stromal cells
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Enzymatic characterization of novel arylsulfatase A variants using human arylsulfatase A-deficient immortalized mesenchymal stromal cells. / Böhringer, Judith; Santer, René; Schumacher, Neele; Gieseke, Friederike; Cornils, Kerstin; Pechan, Maria; Kustermann-Kuhn, Birgit; Handgretinger, Rupert; Schöls, Ludger; Harzer, Klaus; Krägeloh-Mann, Ingeborg; Müller, Ingo.
in: HUM MUTAT, Jahrgang 38, Nr. 11, 31.07.2017, S. 1511-1520.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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T1 - Enzymatic characterization of novel arylsulfatase A variants using human arylsulfatase A-deficient immortalized mesenchymal stromal cells
AU - Böhringer, Judith
AU - Santer, René
AU - Schumacher, Neele
AU - Gieseke, Friederike
AU - Cornils, Kerstin
AU - Pechan, Maria
AU - Kustermann-Kuhn, Birgit
AU - Handgretinger, Rupert
AU - Schöls, Ludger
AU - Harzer, Klaus
AU - Krägeloh-Mann, Ingeborg
AU - Müller, Ingo
N1 - This article is protected by copyright. All rights reserved.
PY - 2017/7/31
Y1 - 2017/7/31
N2 - Metachromatic leukodystrophy (MLD) is an autosomal recessive lysosomal storage disease caused by mutations in the ARSA gene leading to arylsulfatase A (ARSA) deficiency and causing sulfatide accumulation. Main symptoms of the disease are progressive demyelination, neurological dysfunction, and reduced life expectancy. To date, more than 200 different ARSA variants have been reported in MLD patients. Here we report the biochemical characterization of 7 novel pathogenic variants (c.98T > C, c.195delC, c.229G > C, c.545C > G, c.674A > G, c.852T > A, and c.1274A > G), which were found when sequencing a cohort of 31 German MLD families. For that purpose, the ARSA cDNAs carrying the respective mutations inserted by site-directed mutagenesis were cloned into a MigR1 (MSCV, IRES, GFP, retrovirus-1) vector. The constructs were overexpressed using retroviral gene transfer in immortalized, human multipotent mesenchymal stromal cells (MSC) prepared from a patient deficient in ARSA activity (late infantile MLD). In this novel ARSA(-/-) cell system, the seven ARSA mutants showed arylsulfatase A activity of less than 10% when compared to wildtype, which is evidence for the pathogenicity of all 7 variants. In conclusion, the system of ARSA(-/-) immortalized MSC turned out to be a helpful novel tool for the biochemical characterization of ARSA variants. This article is protected by copyright. All rights reserved.
AB - Metachromatic leukodystrophy (MLD) is an autosomal recessive lysosomal storage disease caused by mutations in the ARSA gene leading to arylsulfatase A (ARSA) deficiency and causing sulfatide accumulation. Main symptoms of the disease are progressive demyelination, neurological dysfunction, and reduced life expectancy. To date, more than 200 different ARSA variants have been reported in MLD patients. Here we report the biochemical characterization of 7 novel pathogenic variants (c.98T > C, c.195delC, c.229G > C, c.545C > G, c.674A > G, c.852T > A, and c.1274A > G), which were found when sequencing a cohort of 31 German MLD families. For that purpose, the ARSA cDNAs carrying the respective mutations inserted by site-directed mutagenesis were cloned into a MigR1 (MSCV, IRES, GFP, retrovirus-1) vector. The constructs were overexpressed using retroviral gene transfer in immortalized, human multipotent mesenchymal stromal cells (MSC) prepared from a patient deficient in ARSA activity (late infantile MLD). In this novel ARSA(-/-) cell system, the seven ARSA mutants showed arylsulfatase A activity of less than 10% when compared to wildtype, which is evidence for the pathogenicity of all 7 variants. In conclusion, the system of ARSA(-/-) immortalized MSC turned out to be a helpful novel tool for the biochemical characterization of ARSA variants. This article is protected by copyright. All rights reserved.
KW - Journal Article
U2 - 10.1002/humu.23306
DO - 10.1002/humu.23306
M3 - SCORING: Journal article
C2 - 28762252
VL - 38
SP - 1511
EP - 1520
JO - HUM MUTAT
JF - HUM MUTAT
SN - 1059-7794
IS - 11
ER -