Molecular Therapy. Nucleic Acids最新文献

Antisense oligonucleotides (ASOs) represent an attractive therapeutic approach for CNS disorders. However, ASO-induced neurotoxicity, especially late-onset adverse events, remains a crucial issue, leading to failures in clinical applications. This study aims to determine the neurological features and molecular mechanisms of the late-onset neurotoxicity and provide strategies to overcome this toxicity. We initially established neurobehavioral assays of rodent neurotoxicity with intracerebroventricular and intrathecal injections of various gapmer-type ASOs and a neuronal cytotoxicity analysis. Through both in vitro and in vivo assessments, we identified a site-specific chemical modification, 5'-cyclopropylene (5'-CP), that significantly reduced late-onset neurotoxicity without compromising knockdown activity, providing useful insights into structure-toxicity and structure-activity relationships in ASOs targeting CNS. Additionally, we revealed a toxicity-related mechanism as an elevation of p53-regulated transcripts and paraspeckle protein mislocalization in neuronal cells, which is alleviated through the chemical modifications. Our findings provide mechanistic insights into late-onset ASO-induced neurotoxicity and highlight the potential of optimized chemical modifications to expand the therapeutic window for clinical applications targeting intractable neurological diseases.

Recently, messenger RNA (mRNA)-based vaccine technology has made significant advances in preventing pathogenic microbial infections. The composition and physicochemical properties of lipid nanoparticle (LNP) determine the delivery efficiency of mRNA vaccines. In this study, we synthesized a novel ionizable lipid, C14-192, featuring a 3-oxo-polyamine head group, which was used as a component for LNP to encapsulate and deliver mRNA. Analysis of in vitro and in vivo expression showed that C14-192-LNP-encapsulated luciferase mRNA exhibited high expression efficiency. To further assess the potential of the C14-192 LNP formulation for vaccine applications, we developed a prophylactic mRNA vaccine against Streptococcus pneumoniae (S. pneumoniae), based on the conserved and truncated pneumococcal histidine triad protein D (PhtD) and pneumolysin (Ply). The mRNA encoding the fusion construct exhibited the highest expression and secretion levels. In murine model, mRNA vaccine effectively prevented S. pneumoniae infection and colonization in the lungs and prevented severe lesions. Moreover, the vaccine demonstrated robust cross-protection against multiple serotypes of S. pneumoniae and provide effective protection against lethal infection. In conclusion, a novel ionizable lipid was successfully synthesized and applied in the development of a new prophylactic vaccine against S. pneumoniae.

A significant challenge of mRNA-based protein replacement therapies is the diminishing efficacy and escalating toxicity associated with repeated dosing of lipid nanoparticles (LNPs). Many existing lipid formulations were originally designed for vaccine and are not optimized for therapeutic applications. We developed two libraries of ionizable lipids-one based on piperazine and the other on a newly introduced cyclohexane structure-with variations in linker and tail groups to enhance molecular diversity. GC Biopharma's cyclohexane- and piperazine-based LNPs (GCP LNPs) supported stable and high-level expression without inducing liver toxicity under repeated dosing regimens. These LNPs effectively corrected disease markers in mouse models of phenylketonuria (PKU) and succinic semialdehyde dehydrogenase (SSADH) deficiency. Specially, we observed that the rigid structure and chemical stability of cyclohexane-based lipids contributed to sustained delivery performance. These findings offer a promising direction for the development of LNPs suitable for chronic mRNA-based therapies.

[This corrects the article DOI: 10.1016/j.omtn.2025.102611.].

Loss of integrity of the capillary network is directly associated with the development of kidney fibrosis resulting in chronic kidney disease. Here, we characterized long non-coding RNAs (lncRNAs) in endothelial cells (ECs) during the development of kidney fibrosis. Using a murine EC lineage-tracing model, we observed expression of the conserved lncRNA metastasis-associated lung adenocarcinoma transcript 1 (Malat1) to be elevated in ECs upon kidney injury; either by ischemia-reperfusion injury or by unilateral ureteral obstruction (UUO). In addition, we found elevated MALAT1 expression in the kidney and circulation of patients with fibrotic kidney diseases. Pharmacological intervention of Malat1 initiated protection against fibrosis in the UUO model, illustrated by a marked decline in collagen deposition and a concomitant decrease in interstitial alpha-smooth muscle actin (α-SMA)-positive cells in the kidney. This protective effect was further highlighted by an increase in capillary density and reduced endothelial-to-mesenchymal transition. Mechanistically, transcriptomic analyses of kidney ECs upon Malat1 knockdown demonstrated increased EC-matrix-receptor interaction. Furthermore, we show that silencing of MALAT1 results in increased barrier function and angiogenic response, less vascular leakage, and decreased focal adhesions. Finally, integration with in silico analyses and RNA immunoprecipitation confirmed binding of MALAT1 to SUZ12, a member of the PRC2 complex, suggesting a transcriptional regulatory role for MALAT1. Collectively, our findings classify the lncRNA MALAT1 as an important regulator of EC function and kidney health. As such, targeting MALAT1 may provide novel strategies to reduce kidney fibrosis.