In vitro correction of ARSA deficiency in human skin fibroblasts from metachromatic leukodystrophy patients after treatment with microencapsulated recombinant cells

Abstract

Metachromatic leukodystrophy (MLD) is an autosomal recessive disorder due to arylsulfatase A (ARSA) deficiency that affects primarily the central nervous system. Ongoing treatments include enzyme replacement therapy and bone marrow transplantation, both limited in their effects due to the blood-brain barrier. An alternative approach would be the in situ implantation of encapsulated cells over expressing ARSA. Based on that, we tested the ability of encapsulated BHK cells over expressing ARSA to correct the enzyme deficiency in MLD patients' fibroblasts. Three groups were analyzed: fibroblasts treated with ARSA-over expressing BHK cells (rBHK) trapped in alginate capsules (capsules group), fibroblasts treated with supernatant of non-encapsulated rBHK (uptake control) and fibroblasts treated with empty capsules (empty group). Untreated and normal fibroblasts were used as controls. rBHK obtained by clone selection after non-viral transfection with pTARSA-CMV2. ARSA activity was measured after 1, 2, 3 and 4 weeks of treatment and beta-gal was used as reference enzyme. Statistical analysis was performed using ANOVA and Tukey's test. Normal fibroblasts showed ARSA activity of 23.9 + /- 2.01 nmol/h/mg of protein, whereas untreated MLD fibroblasts had the low ARSA activity (2.22 + /- 0.17). In the empty group, ARSA activity was equal to that of untreated fibroblasts (2.71 + /- 0.34). Capsules and uptake control groups showed higher enzymatic activity levels, compared to MLD untreated, 23.42 + /- 6.39 and 42.35 + /- 5.20, respectively (p < 0.01 for all groups). Encapsulated rBHK clones show potential as a new therapeutic strategy for the treatment of MLD, reaching normal enzyme levels in human MLD fibroblasts.