Vu MN, Neil JA, Mackenzie-Kludas C
… +5 more, Kelly A, Tan HX, Subbarao K, Lee WS, Wheatley AK
Mol Ther Nucleic Acids
· 2026 Jun · PMID 41858837
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Monoclonal antibodies (mAbs) are an emerging class of therapeutics for the prevention and treatment of viral infections. Recent advances in mRNA/lipid nanoparticle (LNP) technology provide a promising new modality for th...Monoclonal antibodies (mAbs) are an emerging class of therapeutics for the prevention and treatment of viral infections. Recent advances in mRNA/lipid nanoparticle (LNP) technology provide a promising new modality for the production of mAbs , potentially bypassing the need for recombinant manufacturing of mAb proteins. In this study, we compared traditional infusion of protein-based neutralizing mAbs targeting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) or influenza viruses to mRNA/LNP-based production of mAbs in treated mice. High serum concentrations of mAbs were achieved upon delivery of a single mRNA encoding both heavy and light chains via intravenous or intramuscular routes using prototypic LNP formulations. However, the pharmacokinetics of mRNA-delivered mAbs were heavily influenced by the induction of anti-drug antibody responses directed against the encoded mAbs, resulting in reduced half-life and compromised protective capacity against SARS-CoV-2 Omicron BA.1 infection. In contrast, mRNA/LNP delivery of a neutralizing mAb conferred superior protection against lethal influenza challenge compared to equivalent recombinant protein doses. Overall, mRNA/LNP delivery comprises a feasible and attractive pathway to speed the development and deployment of antiviral antibodies. However, optimization of LNP formulation, dosing, and administration routes is required to maximize protective potential.
Khan FA, Khan H, Awan UA
… +4 more, Nurahmat M, Nabijan M, Abduwaki M, Dong J
Mol Ther Nucleic Acids
· 2026 Jun · PMID 41858835
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c-Jun, a core component of the activating protein-1 (AP-1) transcription factor complex, regulates cellular processes including proliferation, differentiation, survival, apoptosis, and oncogenesis. c-Jun functions by dim...c-Jun, a core component of the activating protein-1 (AP-1) transcription factor complex, regulates cellular processes including proliferation, differentiation, survival, apoptosis, and oncogenesis. c-Jun functions by dimerizing to bind DNA and modulates the expression of genes such as Bcl-2, cyclin D1, and pro-inflammatory cytokines, enabling context-dependent transcriptional control. Its role in neurodegenerative diseases has gained attention due to its regulation of oxidative stress, inflammation, and apoptosis. In Parkinson's disease, Alzheimer's disease, and Huntington's disease, dysregulated c-Jun expression accelerates dopaminergic neuron loss via oxidative damage, contributes to amyloid-β-induced synaptic toxicity, and mediates neuronal apoptosis and inflammation, respectively. Despite its degenerative role, c-Jun also promotes axonal regeneration and stress adaptation, revealing a dual function that depends on context and stimulus severity. This paradox underscores its ability to promote survival under mild stress and apoptosis under chronic damage. Emerging therapeutic strategies targeting c-Jun-via small-molecule inhibitors (e.g., SP600125), RNA interference, or modulation of upstream c-Jun N-terminal kinase (JNK)-are being explored. However, challenges remain in achieving specificity, as c-Jun's ubiquitous expression raises concerns about off-target effects. This review highlights recent advances in understanding c-Jun's complex role in neurodegeneration and its therapeutic potential, emphasizing its value as both a mechanistic regulator and a target for preserving neuronal integrity in neurodegenerative diseases.
Kahwaji N, Kotzian N, Prinz JM
… +17 more, Pu Y, Kath J, Picht S, Hiller AL, Klaas A, Dunne CM, Launspach M, Palmowski A, Kleyer A, Simon DN, Wagner DL, Pichon C, Krönke G, Schmueck-Henneresse M, Volk HD, Gossen M, Drzeniek NM
Mol Ther Nucleic Acids
· 2026 Mar · PMID 41834823
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Unmodified, uridine-containing mRNA is known to trigger antiviral immune responses, inflammatory signaling, and apoptosis in transfected cells. To avoid this and enable high expression, modified nucleosides such as N1-me...Unmodified, uridine-containing mRNA is known to trigger antiviral immune responses, inflammatory signaling, and apoptosis in transfected cells. To avoid this and enable high expression, modified nucleosides such as N1-methylpseudouridine have become the gold standard for mRNA applications including T cell engineering, albeit at increased cost. Here, immune responses toward mRNA were evaluated across five primary human cell types. Remarkably, T cells, unlike other immune and non-immune cell types tested, exhibited no immune activation by unmodified mRNA. T cell viability and cytokine secretion remained unaffected, regardless of mRNA delivery method via lipid nanoparticles or electroporation. The absence of nucleotide modifications improved expression of chimeric antigen receptor (CAR) in activated T cells and CAR-T cell cytotoxic potency. By eliminating the need for mRNA-nucleoside modification in CAR-T cell engineering, our findings challenge existing paradigms and position mRNA as a non-inflammatory, minimally invasive and highly efficient tool for T cell engineering, while simplifying and reducing manufacturing cost.
Martelli C, Bonanomi M, Pellizzer C
… +8 more, Giammona A, Remedia S, Nespoli M, Gaglio D, Capitanio D, Porro D, Ottobrini L, Lo Dico A
Mol Ther Nucleic Acids
· 2026 Mar · PMID 41834822
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Glioma is currently the most aggressive CNS tumor. MicroRNA (miRNA)675-5p is a hypoxic miRNA involved in promoting and maintaining the hypoxia inducible factor (HIF)-1α pathway, the driving force for glioma proliferation...Glioma is currently the most aggressive CNS tumor. MicroRNA (miRNA)675-5p is a hypoxic miRNA involved in promoting and maintaining the hypoxia inducible factor (HIF)-1α pathway, the driving force for glioma proliferation, migration into surrounding tissue, and resistance. Its inhibition is effective in reducing HIF-1α and all pathways related to it. However, the molecular mechanism through which miRNA inhibition is effective has not yet been fully elucidated. The therapeutic efficacy of the miRNA675-5p inhibitor was tested in a panel of resistant glioma lines evaluating the cellular, molecular, and biochemical rearrangement of the cells after treatment, with particular attention to the oxidative stress imbalance. miRNA675-5p inhibitor has a therapeutic efficacy on its own in resistant cell lines, reducing HIF-1α and its related pathways. The mechanism through which this occurs is the induction of oxidative stress. Its impairment, in fact, reverses the cytotoxic effect. Inhibitor-treated cells acquire metabolic characteristics clearly distinct from untreated cells, triggering compensatory mechanisms that must be considered for the secondary treatment of glioma with temozolomide. The induction of oxidative stress and metabolic rearrangement play key roles in the cytotoxic effect of the miRNA675-5p inhibitor, which could be proposed as a new therapeutic approach in glioma.
Song P, Su J, Kristine Vraa CL
… +4 more, Gockert M, Fjelstrup S, Hager H, Kjems J
Mol Ther Nucleic Acids
· 2026 Mar · PMID 41815890
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Lipid nanoparticles (LNPs) have proven to be an effective delivery system for RNA therapeutics. The chemical composition of LNPs determines their functional delivery efficiency and targeting properties, which vary betwee...Lipid nanoparticles (LNPs) have proven to be an effective delivery system for RNA therapeutics. The chemical composition of LNPs determines their functional delivery efficiency and targeting properties, which vary between and contexts. Here, we have systematically characterized and compared 25 novel C14120-based LNP formulations for mRNA delivery and assessed mRNA expression and biodistribution using deep sequencing of DNA barcodes in a pooled LNP-mRNA library. experiments showed correlations of lipid composition with particle size and mRNA transfection efficiency in 4 different cell lines of distinct tissue and species origin. experiments employed a pooled LNP delivery of luciferase mRNA in combination with a multiplexed barcode system and identified LNP compositions with organ-specific targeting properties. Individual validation of three selected LNP candidates based on mRNA expression analysis confirmed high specificity for the lung-targeting candidate, lower specificity for the liver-targeting candidate, and inconclusive results for the spleen-targeting candidate. These findings identify LNP formulations with promising potential for and organ-targeted delivery.
Mol Ther Nucleic Acids
· 2026 Mar · PMID 41810142
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Lymphotoxin alpha (LTα) is a potent inflammatory cytokine implicated in the pathophysiology of numerous human autoimmune and inflammatory diseases. Existing as a soluble homotrimer (LTα) or membrane-bound heterotrimer (L...Lymphotoxin alpha (LTα) is a potent inflammatory cytokine implicated in the pathophysiology of numerous human autoimmune and inflammatory diseases. Existing as a soluble homotrimer (LTα) or membrane-bound heterotrimer (LTαβ), the differential and distinct functions of lymphotoxin signaling have meant that selective targeting of the cytokine with traditional pharmacological agents has proven difficult. While monoclonal antibodies that can neutralize human LTα do exist (e.g., pateclizumab), their efficacy and subsequent use within the clinic have been limited. This may be in part perhaps due to cross-reactivity between the homotrimeric and heterotrimeric forms, leading to LTα-independent effects. Herein, we implement a counter-Systematic Evolution of Ligands by Exponential Enrichment (SELEX) protocol to enrich aptamers targeting LTα but not LTαβ Through a combination of and tests, we also refine the aptamer sequences and test their ability to limit LT⍺-TNFR1 engagement and subsequent cellular cytotoxicity in L929 fibroblasts. We highlight the generation of 4 aptamer candidates that can selectively detect LTα but not LTαβ. Using rational design, we optimize the sequences and show that LTa1 and LTa5 can significantly reduce LTa-TNFR1 engagement-associated cytotoxicity , highlighting their future therapeutic potential. These data highlight aptamers for the future investigation of lymphotoxin signaling, limiting off-target impacts on other LT⍺-dependent mechanisms.
Mol Ther Nucleic Acids
· 2026 Mar · PMID 41810141
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This study highlights that small geometric constraints with the GalNAc moiety can tune receptor engagement and serum stability. The ribofuranose ring is a 2'-O-methyl modification, used in the siRNA backbone to improve m...This study highlights that small geometric constraints with the GalNAc moiety can tune receptor engagement and serum stability. The ribofuranose ring is a 2'-O-methyl modification, used in the siRNA backbone to improve metabolic stability and reduce nuclease susceptibility. The constrained ribofuranose conformation introduced into GalNAc increased serum stability and hepatic parenchymal clearance, thereby increasing systemic exposure. They also demonstrate the kilogram-scale synthesis of the G5 GalNAc-CPG support, making it feasible for manufacturing. PCSK9 is an ideal benchmark target as it is clinically validated, and the observed performance can be applied to other hepatocyte-expressed genes. The inc-G5 GalNAc has progressed to phase 1 trials in China for PCSK9- and AGT-targeting siRNA. In a non-human primate study, inc-G5 GalNAc-siRNA conjugates showed a safety profile consistent with established GalNAc-siRNA (inc-L96) conjugates. A comparison of toxicology in AGT-targeting siRNA versus zilebesiran showed similar clinical chemistry and histopathology, supporting inc-G5 GalNAc as a potent, clinically translatable GalNAc delivery scaffold.
Mu J, Wang C, Shu T
… +5 more, Xiong X, Ren Y, Gong R, Zhao L, Zhou X
Mol Ther Nucleic Acids
· 2026 Mar · PMID 41783789
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Rabies lyssavirus (RABV) is a prototypical neurotropic virus that belongs to the Rhabdoviridae family and causes one of the most lethal zoonotic diseases. For RNA viruses, RNA-unwinding proteins such as helicases and RNA...Rabies lyssavirus (RABV) is a prototypical neurotropic virus that belongs to the Rhabdoviridae family and causes one of the most lethal zoonotic diseases. For RNA viruses, RNA-unwinding proteins such as helicases and RNA chaperones are crucial for the replication and/or correct folding of viral RNAs. Although numerous RNA viruses have been found to encode their RNA-unwinding proteins, no RNA-unwinding activity has been identified in the family Rhabdoviridae. Herein, we uncovered that RABV phosphoprotein (RABV-P) possesses both a nucleoside triphosphate (NTP)-dependent helicase-like RNA duplex-unwinding activity and an NTP-independent duplex-unwinding and annealing activity. The C-terminal domain of RABV-P is required for its RNA-unwinding activities, whereas the NTP-dependent helicase-like unwinding activity also relies on its central oligomerization domain (COD). Furthermore, we designed two peptides (named P90 and P110) to target the COD and revealed that these peptides effectively inhibited RABV replication in cells. Overall, the study provides the first demonstration of the RNA helicase-like and NTP-independent RNA-unwinding/annealing activities associated with a rhabdovirus-encoded protein. This study highlights the functional significance of P in the RABV life cycle and probably other rhabdoviruses, and targeting RABV-P represents a promising strategy for developing novel antivirals against this lethal pathogen.
Khorsandi SE, Vasconcelos D, Nicholas R
… +9 more, Reebye V, Nicholls J, Sodergren M, Rowell J, Dehkordi A, Habib N, Swiderski P, Rossi J, Huang KW
Mol Ther Nucleic Acids
· 2026 Mar · PMID 41783787
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CCAAT/enhancer-binding protein beta (C/EBPβ) is a master regulator of hepatic metabolism, inflammation, and fibrosis, making it an attractive but underexploited target for metabolic dysfunction-associated steatotic liver...CCAAT/enhancer-binding protein beta (C/EBPβ) is a master regulator of hepatic metabolism, inflammation, and fibrosis, making it an attractive but underexploited target for metabolic dysfunction-associated steatotic liver disease (MASLD). Here, we demonstrate that GalNAc-conjugated small interfering RNA (siRNA) targeting C/EBPβ (GalNAc-siCEBPβ) significantly improves liver function and metabolic parameters in a high fat diet (HFD) murine model. , GalNAc-siCEBPβ achieved dose-dependent C/EBPβ mRNA silencing (∼80% knockdown at 0.1 μM) in primary mouse hepatocytes. , subcutaneous administration (10 mg/kg) reduced hepatic C/EBPβ expression by 45% ( < 0.01), concomitant with a marked reduction in liver steatosis and improved metabolic profile (15% less weight gain, 20% lower glucose, 25% reduced triglycerides), and restored liver function (18% higher albumin, 22% lower bilirubin)-all without hepatotoxicity (ALT/AST unchanged). Notably, these effects occurred despite continued HFD feeding, suggesting disease-modifying potential. By leveraging the precision of RNAi and hepatocyte-specific GalNAc delivery, GalNAc-siCEBPβ addresses key limitations of current MASLD therapies by targeting both metabolism and fibrosis. Our findings support clinical translation for MASLD and its complications, including hepatocellular carcinoma.
Porter D, Jouneau L, Peruzzi M
… +3 more, Collet B, Verrier B, Boudinot P
Mol Ther Nucleic Acids
· 2026 Mar · PMID 41757374
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Messenger RNA (mRNA) vaccines have revolutionized immunization strategies in mammals, with chemical modifications such as N1-methyl-pseudouridine (N1MΨU) enhancing stability, translation, and reducing innate immune activ...Messenger RNA (mRNA) vaccines have revolutionized immunization strategies in mammals, with chemical modifications such as N1-methyl-pseudouridine (N1MΨU) enhancing stability, translation, and reducing innate immune activation. However, the immunological implications of such modifications in non-mammalian species, particularly teleost fish, remain unclear. In this study, we evaluated the innate immune responses elicited by four vaccine platforms: a DNA vaccine, a live attenuated viral hemorrhagic septicemia virus (VHSV), an unmodified mRNA, and an N1MΨU-modified mRNA. All four vaccines encode the G protein of VHSV (G) in rainbow trout. Following intramuscular injection, both mRNA vaccine formats induced robust type I interferon (IFN) responses, comparable to those induced by the DNA and attenuated virus vaccines. Notably, the N1MΨU-modified mRNA vaccine did not suppress IFN induction, as observed in mammalian systems, but instead triggered distinct temporal transcriptional dynamics and stronger enrichment of autophagy, ubiquitination, and transcription-related pathways. These findings suggest that modified mRNA retains a strong immunostimulatory capacity in fish and may activate immune pathways differently than in mammals. Our results emphasize the need to reassess assumptions from mammalian vaccine design when translating mRNA vaccine strategies to non-mammalian vertebrates and highlight the potential for tailored mRNA vaccine development in aquaculture species.