Molecular Therapy. Nucleic Acids最新文献

Variants in KIAA0586/TALPID3 are associated with the ciliopathy Joubert syndrome (JS), which is a genetically heterogeneous disorder. Mutations in more than 40 different genes were associated with JS, genes that are relevant to ciliary assembly, maintenance, or cellular signaling pathways during the development. Genetic variability suggests that gene- and/or mutation-independent treatment strategies could be beneficial to patients. Currently, targeted therapeutic options are not available. In this study, we present molecular treatment options for pathogenic KIAA0586/TALPID3 sequence alterations. We compared therapeutic efficacy and side effects using patient-derived fibroblasts from two siblings affected by JS. The patients harbored two compound heterozygous sequence alterations, a non-sense mutation (KIAA0586/TALPID3: c.2353C>T) in exon 18 and a deep-intronic mutation in intron 28 (KIAA0586/TALPID3: c.3990 + 3186G>A). The deep-intronic sequence alteration activates a cryptic exon that causes a frameshift and splicing defect. Both variants are predicted to potentially result in the premature termination of translation. The patient-derived fibroblasts exhibited reduced primary cilia length and altered distribution of PCM1. These cellular defects were responsive to treatments with RNA-based therapeutics and/or readthrough agents (RTAs). Our results highlight the potential of addressing mutations and molecular defects associated with KIAA0586/TALPID3 sequence alterations as future perspectives toward treatments of patients.

Circular RNAs (circRNAs) are a class of RNA molecules with a covalent closed-loop structure that play important roles in the regulation of biological processes and disease genesis. In recent years, circRNAs have become a research hotspot due to their unique stability and biological functions. Compared with linear RNAs, synthetic circRNAs have higher stability and show great potential in therapeutic applications. Although circRNA technology is still at an early stage of development, breakthroughs in mRNA technology provide an important reference for its application. This review systematically explores the promising applications of circRNAs in vaccine development and drug research, evaluates their feasibility as vaccine components and drug carriers, and experimentally validates their efficacy in disease models. At the same time, this paper analyzes the advantages and challenges of circRNA application in depth and looks forward to the future research direction, which provides new ideas for the prevention and treatment of new outbreaks of infectious diseases.

Significant progress has been made in the development of lipid nanoparticle (LNP)-encapsulated messenger RNA (mRNA)-based modalities with numerous candidates entering clinical trials. These novel modalities require the application of innovative quantitative models to inform the development of mRNA-LNP-based candidates. To this end, we conducted a comprehensive search of registered clinical trials related to mRNA-based modalities on ClinicalTrials.gov, summarizing the current advancements of mRNA-LNP-based modalities and their expanding therapeutic applications. Also, we performed a thorough review of quantitative models related to mRNA-LNP-based modalities from PubMed, Google Scholar, and Embase databases, exploring the model structures employed to capture the in vivo processes of mRNA-LNP, along with their current applications. Between 2002 and October 28, 2024, around 189 clinical trials were registered on ClinicalTrials.gov, encompassing approximately 132 unique mRNA-based modalities targeting 18 disease areas. There are 15 studies that have published quantitative models supporting both the preclinical and clinical development of mRNA-LNP-based therapeutics. Detail regarding quantitative modeling of mRNA-LNP, especially absorption, distribution, metabolism, and excretion, as well as the activation processes of immune responses induced by mRNA-LNP-based vaccines are reviewed. Furthermore, we offer insights for future research related to mRNA-LNP-related models, aimed at enhancing predictive performance and facilitating the expedited advancement of mRNA-LNP-related clinical development.

We combined circular dichroism (CD) and viscosity measurements with molecular dynamics (MD) simulations and classification and regression approaches to machine learning to characterize solution structures of 22-mer, 25-mer, and 30-mer peptide- (-GlyArg6) conjugated phosphorodiamidate morpholino oligonucleotides (PPMOs). PPMO molecules form non-canonical folded structures with 1.4- to 1.5-nm radius of gyration, 4-6 base pairs and 5-11 base stacks, characterized by -49 to -71 kcal/mol free energy of folding. The 4.5-6.1 cm³/g intrinsic viscosity and Huggins constant of 4.5-9.7 indicate PPMO-PPMO interactions at higher concentrations. The random-coil 3'-end conjugated -GlyArg₆ portion does not alter molecular properties of phosphorodiamidate morpholino oligonucleotide (PMO) components, which explains why CD spectra, viscosity-concentration profiles, and inhibitor activities of 22-mer, 25-mer, and 30-mer PPMOs and PMOs are similar but the peptide enhances the PPMO cellular uptake. PPMOs' viscosity is lower than PMOs' viscosity, due to PMO-peptide position-dependent interactions, especially in 25-mer PPMO, explaining differences in CD and high-concentration viscosity. These results reiterate the importance of the conformational ensemble view of non-canonical PPMO structures in solution, in agreement with our previous PMO study. The addition of -GlyArg₆ does not alter the structure and molecular properties of the PMO components of the PPMO structures but impacts the viscosity of the PPMO-based aqueous solution formulations.

Duchenne muscular dystrophy (DMD) is the most common inherited muscle disease. There are currently few effective therapies to treat the disease, although many approaches are being pursued. Certain histone deacetylase inhibitors (HDACi) have been shown to ameliorate DMD phenotypes in mouse and zebrafish models, and the HDACi givinostat has recently gained FDA approval for DMD. Our goal was to identify additional HDACi, or other classes of epigenetic small molecules, that are beneficial for DMD. Using an established animal model for DMD, the zebrafish dmd mutant strain sapje, we screened a library of over 800 epigenetic small molecules. Our screening identified a new HDACi, SR-4370, that ameliorated dmd mutant zebrafish skeletal muscle degeneration, as well as additional HDACi that have previously been shown to improve dmd zebrafish. We find that a single early treatment of HDACi can ameliorate the muscle phenotype and increase lifespan in dmd zebrafish. Furthermore, we find that HDACi treatments that improve dmd muscle also cause increased histone acetylation in zebrafish larvae. Our results add to the growing evidence that HDACi are promising candidates for treating DMD. Our study also provides further support for the effectiveness of small molecule screening in dmd zebrafish.