摘要：SummaryWe show the successful application of ancestral sequence reconstruction to enhance the activity of iduronate-2-sulfatase (IDS), thereby increasing its therapeutic potential for the treatment of Hunter syndrome—a lysosomal storage disease caused by impaired function of IDS. Current treatment, enzyme replacement therapy with recombinant human IDS, does not alleviate all symptoms, and an unmet medical need remains. We reconstructed putative ancestral sequences of mammalian IDS and compared them with extant IDS. Some ancestral variants displayed up to 2-fold higher activity than human IDS inin vitroassays and cleared more substrate inex vivoexperiments in patient fibroblasts. This could potentially allow for lower dosage or enhanced therapeutic effect in enzyme replacement therapy, thereby improving treatment outcomes and cost efficiency, as well as reducing treatment burden. In summary, we showed that ancestral sequence reconstruction can be applied to lysosomal enzymes that function in concert with modern enzymes and receptors in cells.Graphical abstractDisplay OmittedHighlights•Reconstruction of ancestral lysosomal enzymes that function in complex cellular context•Ancestral iduronate-2-sulfatases with increased activity compared with the human enzyme•Increased clearance of substrate in patient fibroblasts indicates therapeutic potentialBiochemistry; Structural Biology
