Ancestral lysosomal enzymes with increased activity harbor therapeutic potential for treatment of Hunter syndrome

Abstract

Summary We 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 in in vitro assays and cleared more substrate in ex vivo experiments 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.

Degradation of PET microplastic particles to monomers in human serum by PETase

“…Enzymes have consolidated themselves as an essential tool in the biopharmaceutical industry [34][35][36] and, therefore, we sought to test the potential of IsPETase (WT PETase) and its S238A variant to convert microplastic in human serum. It is well known that many enzymes have a preferred pH and salinity to operate in.…”

Pseudo-perplexity in One Fell Swoop for Protein Fitness Estimation

“…The technique has made it possible to resurrect proteins that may have existed eons ago, and thereby directly investigate the functional properties of these inferred ancestral sequences [34][35][36][37][38][39]. The research program has led to the rather perplexing finding that reconstructed ancestral sequences tend to have superior functional properties compared to their extant counterparts: improved activity, enhanced stability and/or higher promiscuity [40][41][42][43]. This is true, so much so that, ancestral state reconstruction has emerged as a viable strategy for protein design [44,45].…”

Engineering Synthetic and Recombinant Human Lysosomal b-Glucocerebrosidase for Enzyme Replacement Therapy for Gaucher Disease

“…These engineered enzymes have also been used as a drug-resistance gene to optimize lentiviral vectors for HIV and sickle disease gene therapy [42,43]. Additionally, engineered enzymes such as iduronate-2-sulfatase (IDS) have been developed for the treatment of Hunter syndrome-α lysosomal storage disease [44], and α-galactosidase (GLA) for the treatment of Fabry disease [45]. Furthermore, researchers have used synthetic biology technologies for programming, controlling transgene expression (35,36), and for synthesizing codon-optimized DNA constructs for gene therapy for several genetic diseases [46][47][48].…”