Enzymatic characterization of novel arylsulfatase A variants using human arylsulfatase A-deficient immortalized mesenchymal stromal cells

TY - JOUR

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 -