Nucleic acid‐based therapeutics that regulate gene expression have been developed towards clinical use at a steady pace for several decades, but in recent years the field has been accelerating. To date, there are 11 marketed products based on antisense oligonucleotides, aptamers and small interfering RNAs, and many others are in the pipeline for both academia and industry. A major technology trigger for this development has been progress in oligonucleotide chemistry to improve the drug properties and reduce cost of goods, but the main hurdle for the application to a wider range of disorders is delivery to target tissues. The adoption of delivery technologies, such as conjugates or nanoparticles, has been a game changer for many therapeutic indications, but many others are still awaiting their eureka moment. Here, we cover the variety of methods developed to deliver nucleic acid‐based therapeutics across biological barriers and the model systems used to test them. We discuss important safety considerations and regulatory requirements for synthetic oligonucleotide chemistries and the hurdles for translating laboratory breakthroughs to the clinic. Recent advances in the delivery of nucleic acid‐based therapeutics and in the development of model systems, as well as safety considerations and regulatory requirements for synthetic oligonucleotide chemistries are discussed in this review on oligonucleotide‐based therapeutics.

Impaired degradation of glycosaminoglycans (GAGs) with consequent intralysosomal accumulation of undegraded products causes a group of lysosomal storage disorders known as mucopolysaccharidoses (MPSs). Characteristically, MPSs are recognized by increased excretion in urine of partially degraded GAGs which ultimately result in progressive cell, tissue, and organ dysfunction. There are eleven different enzymes involved in the stepwise degradation of GAGs. Deficiencies in each of those enzymes result in seven different MPSs, all sharing a series of clinical features, though in variable degrees. Usually MPS are characterized by a chronic and progressive course, with different degrees of severity. Typical symptoms include organomegaly, dysostosis multiplex, and coarse facies. Central nervous system, hearing, vision, and cardiovascular function may also be affected. Here, we provide an overview of the molecular basis, enzymatic defects, clinical manifestations, and diagnosis of each MPS, focusing also on the available animal models and describing potential perspectives of therapy for each one.

Lysosomal storage diseases (LSDs) are a group of rare, life-threatening genetic disorders, usually caused by a dysfunction in one of the many enzymes responsible for intralysosomal digestion. Even though no cure is available for any LSD, a few treatment strategies do exist. Traditionally, efforts have been mainly targeting the functional loss of the enzyme, by injection of a recombinant formulation, in a process called enzyme replacement therapy (ERT), with no impact on neuropathology. This ineffectiveness, together with its high cost and lifelong dependence is amongst the main reasons why additional therapeutic approaches are being (and have to be) investigated: chaperone therapy; gene enhancement; gene therapy; and, alternatively, substrate reduction therapy (SRT), whose aim is to prevent storage not by correcting the original enzymatic defect but, instead, by decreasing the levels of biosynthesis of the accumulating substrate(s). Here we review the concept of substrate reduction, highlighting the major breakthroughs in the field and discussing the future of SRT, not only as a monotherapy but also, especially, as complementary approach for LSDs.

Background The genetic polymorphism of thiopurine methyltransferase (TPMT) activity has a significant relevance in the clinical outcome of patients receiving thiopurine drugs as immunosupressive or anticancer therapies. Apart from several open reading frame mutations unequivocally associated with decreased TPMT activity, a variable number of tandem repeats (VNTR), located within the 5′ untranslated region, was recently reported as also affecting gene expression. Aims and Methods We have characterized both molecularly, by polymerase chain reaction–based techniques, and enzymatically, with an HPLC‐based method, a sample of 143 Portuguese Caucasian individuals with the main objective of deepening the study of the TPMT genotype/phenotype relationship. Because two different repeated elements (A and B) do contribute to the overall VNTR variation, we set out to analyze their combined and individual effects on TPMT activity. Results Allele VNTR*6 was found to be consistently associated with decreased levels of TPMT activity, supporting previous reports that the VNTR does affect levels of TPMT activity, although moderately and in a way not yet clearly defined. Furthermore, individual analysis of the A and B variations suggested that three B repeats was the likely motif influencing gene expression toward decreased transcription. Conclusions Our hypothesis, which emphasizes the number of particular motifs within the VNTR internal structure more than the undiscriminated number of repeats as the potential causative factor affecting TPMT activity, needs further support from future studies, namely, from enlarged population samples. However, the knowledge of the VNTR acting mechanism will represent an important step toward fully understanding how the phenotypic variability at the TPMT trait is modulated. Clinical Pharmacology & Therapeutics (2001) 70, 165–174; doi: 10.1067/mcp.2001.117284