Abstract
Gaucher disease is an autosomal recessive disorder caused by deficiency of the enzyme glucocerebrosidase. Although it is a monogenic disease, there is vast phenotypic heterogeneity, even among patients with the same genotype. MicroRNAs (miRNAs) are small non-coding RNAs involved in many biological processes and diseases. To determine whether miRNAs can affect glucocerebrosidase activity, we performed a screen of 875 different miRNA mimics. The screen was performed using Gaucher fibroblasts, and glucocerebrosidase activity was used as the initial outcome parameter. We found several miRNAs that either up- or down-regulated glucocerebrosidase activity. In follow-up assays, we confirmed that one specific miRNA (miR-127–5p) down-regulated both glucocerebrosidase activity and protein levels by down-regulation of LIMP-2, the receptor involved in proper trafficking of glucocerebrosidase from the endoplasmic reticulum to the lysosome. A conditioned media assay demonstrated that cells treated with this miRNA secreted glucocerebrosidase into the extracellular environment, supporting impaired LIMP-2 function. Two other miRNAs, miR-16–5p and miR-195–5p, were found to up-regulate glucocerebrosidase activity by greater than 40% and to enhance expression and protein levels of the enzyme. In conclusion, we show that miRNAs can alter glucocerebrosidase activity in patient cells, indicating that miRNAs can potentially act as modifiers in Gaucher disease.
Highlights
Gaucher disease (GD) is the most common lysosomal storage disorder worldwide
We found that GCase activity can be altered by select miRNA mimics, and that some of these effects can be explained by the down- or up-regulation of GBA1 mRNA levels, GCase protein levels and/or by affecting other proteins related to GCase, such as LIMP-2
Results miRNA screening, hit selection, and reconfirmation In the present study, miRNA mimic screening (Sanger miRBase 13.0) was performed in wild type (WT) and N370S/N370S Gaucher fibroblasts to evaluate the effects of introducing different miRNAs in increased abundance on GCase activity
Summary
The disease frequency has been estimated between 1:40,000 and 1:60,000 individuals in the general population, while it has a higher frequency in the Ashkenazi Jewish population (1:850 individuals).[1,2] GD, an autosomal recessive disorder, has been clinically subdivided according to severity of symptoms and involvement of the Central Nervous System (CNS).[1] Clinical manifestations in patients with type 1 or nonneuronopathic GD (OMIM #230800) range from asymptomatic individuals to children with significant hepatosplenomegaly, anemia, thrombocytopenia, and bone disease. Type 2 or acute neuronopathic GD (OMIM #230900) is the most severe form, leading to a very short life expectancy (less than 2 years) due to devastating and rapidly progressive neurological impairment. Patients with type 3 GD have CNS involvement, these symptoms often develop later, and the progression of the disease is less aggressive.
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