Molecular Therapy. Nucleic Acids最新文献_第2页

Sub-stoichiometric 5-methoxyuridine modification enables tunable immune evasion and protein expression from synthetic mRNAs. 亚化学计量的5-甲氧基尿嘧啶修饰可以调节合成mrna的免疫逃避和蛋白质表达。

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2026-08-12 eCollection Date: 2026-09-08 DOI: 10.1016/j.omtn.2026.103058

Jonathan R Miles, Sarah Masterson, Eleanor Bellows, Fabio Fisher, Ritu Rani, Sara Marelli, Colin Hardman, Luigi Grassi, Luis Santos, Victoria James, Nigel P Mongan, Christopher J Hayes, Matthew Loose, Janet M Daly, James Button, George Thom, Rupert G Fray, Nathan Archer

{"title":"Sub-stoichiometric 5-methoxyuridine modification enables tunable immune evasion and protein expression from synthetic mRNAs.","authors":"Jonathan R Miles, Sarah Masterson, Eleanor Bellows, Fabio Fisher, Ritu Rani, Sara Marelli, Colin Hardman, Luigi Grassi, Luis Santos, Victoria James, Nigel P Mongan, Christopher J Hayes, Matthew Loose, Janet M Daly, James Button, George Thom, Rupert G Fray, Nathan Archer","doi":"10.1016/j.omtn.2026.103058","DOIUrl":"10.1016/j.omtn.2026.103058","url":null,"abstract":"mRNA therapeutics depend on nucleotide modification to evade innate immunity and sustain translation. Current clinical designs replace all uridine with N1-methylpseudouridine (m1Ψ). However, full substitution is costly and can perturb translational fidelity and RNA structure. Here, we show that sub-stoichiometric incorporation of 5-methoxyuridine (5moU) is sufficient to confer immune evasion while enhancing protein output. Substitution of 25% of uridines with 5moU supported translation comparable to fully m1Ψ-modified mRNA in some cell types, while 50% 5moU maximized expression across all systems tested, often outperforming 100% m1Ψ-modified mRNA. This effect was conserved in human primary cells and in vivo, where lipid nanoparticle-delivered mRNA encoding human IgG produced approximately 2-fold higher serum levels with 50% 5moU than with 100% m1Ψ. Transcriptomic profiling revealed reduced interferon-stimulated gene induction following delivery of 5moU-modified mRNA compared with unmodified and m1Ψ modified mRNA. Unlike m1Ψ, 5moU modification consistently suppressed innate responses to double-stranded RNA. Together, these data demonstrate that partial nucleotide substitution can outperform current mRNA modification strategies and expand the design space for therapeutic mRNAs.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"37 3","pages":"103058"},"PeriodicalIF":6.5,"publicationDate":"2026-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13543807/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148897709","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Adenovirus vector-mediated delivery of the CRISPR-CasRx system enables gene knockdown with suppressed collateral activity. 腺病毒载体介导的CRISPR-CasRx系统的递送使基因敲低与抑制侧枝活性。

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2026-08-12 eCollection Date: 2026-09-08 DOI: 10.1016/j.omtn.2026.103057

Yuki Osakada, Sota Kidawara, Eiko Sakai, Akira Asayama, Fuminori Sakurai, Kahori Shimizu, Hiroyuki Mizuguchi

{"title":"Adenovirus vector-mediated delivery of the CRISPR-CasRx system enables gene knockdown with suppressed collateral activity.","authors":"Yuki Osakada, Sota Kidawara, Eiko Sakai, Akira Asayama, Fuminori Sakurai, Kahori Shimizu, Hiroyuki Mizuguchi","doi":"10.1016/j.omtn.2026.103057","DOIUrl":"10.1016/j.omtn.2026.103057","url":null,"abstract":"The CRISPR-Cas13 system is a programmable gene knockdown tool widely used in basic and clinical research. Meanwhile, RNA-targeting by Cas13 exhibits collateral activity, i.e., cleavage of non-target RNAs, raising serious concerns. Accumulating evidence suggests the collateral activity comes from high expression levels of the Cas13 system. Therefore, controlled expression of the Cas13 system using viral vectors could solve the problem. Adenovirus vectors are efficient gene delivery vehicles and commonly used for clinical therapy. Here, we generated adenovirus vectors carrying a CRISPR-Cas13 system using CasRx (Ad-CasRx). Ad-CasRx efficiently knocked down the on-target reporter with suppressed collateral activity in cell lines, while plasmid transfection-mediated delivery exhibited strong collateral activity. Moreover, Ad-CasRx efficiently knocked down Pcsk9 in cultured murine hepatic cells without suppressing abundant endogenous RNAs, suggesting that collateral activity was largely suppressed when targeting endogenous genes with Ad-CasRx. Finally, we targeted hepatic Pcsk9 in mice by intravenous administration of Ad-CasRx. Pcsk9 mRNA and serum total cholesterol levels were reduced without affecting the expression of major genes related to cholesterol metabolism and hepatic function. Serum ALT levels were not elevated. Overall, controlled delivery of Ad-CasRx enabled in vivo gene knockdown without evident side effects from collateral activity or Cas13 expression itself in liver.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"37 3","pages":"103057"},"PeriodicalIF":6.5,"publicationDate":"2026-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13560539/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148944667","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

To scramble or not: Steric blocking 2'-MOE/PS control antisense oligonucleotides across cellular models. 混乱与否：空间阻断2'-MOE/PS控制反义寡核苷酸跨越细胞模型。

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2026-08-04 eCollection Date: 2026-09-08 DOI: 10.1016/j.omtn.2026.103051

Edith De Bruycker, Manon Bouckaert, Kyana Van Acker, Emma Delanote, Leander Meuris, Álvaro González-Cid, Hanne Lenaerts, Lode Denolf, Katrien Remaut, An Staes, Kris Gevaert, Steve Lefever, Frauke Coppieters

{"title":"To scramble or not: Steric blocking 2'-MOE/PS control antisense oligonucleotides across cellular models.","authors":"Edith De Bruycker, Manon Bouckaert, Kyana Van Acker, Emma Delanote, Leander Meuris, Álvaro González-Cid, Hanne Lenaerts, Lode Denolf, Katrien Remaut, An Staes, Kris Gevaert, Steve Lefever, Frauke Coppieters","doi":"10.1016/j.omtn.2026.103051","DOIUrl":"10.1016/j.omtn.2026.103051","url":null,"abstract":"Antisense oligonucleotides (ASOs) are emerging as a promising therapeutic approach for many disorders. Currently, fourteen ASOs have been FDA/EMA-approved, with many more in preclinical development. An important requirement for preclinical studies is the inclusion of negative controls to properly assess the efficacy of candidate ASOs. As commonly used scrambled or sense ASOs may elicit unwanted off-target effects that could influence downstream data analysis, there is a high need for the inclusion of additional negative control ASOs (cASOs) that are well-characterized and well-tolerated. In this study, we designed and evaluated generic cASOs across seven different cellular models. Characterization at the RNA and protein level revealed potential ASO toxicity mechanisms as well as species-specific and cell-type-specific effects. In total, three steric blocking 2'-MOE/PS cASOs with a distinct GC content were identified that exert limited off-target effects across all cellular models. To conclude, we report a set of cASOs that are potentially applicable in a broad variety of experiments using ASOs with similar physicochemical properties and across different cellular models.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"37 3","pages":"103051"},"PeriodicalIF":6.5,"publicationDate":"2026-08-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13571576/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148963628","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Simultaneous quantitative detection of multiple low-frequency variants by high-dynamic-range capillary electrophoresis. 高动态范围毛细管电泳同时定量检测多种低频变异。

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2026-07-30 eCollection Date: 2026-09-08 DOI: 10.1016/j.omtn.2026.103042

Yoshihiko Hagiwara, Nobue Tamamura, Noriyuki Sumida, Yusuke Ono, Kenji Takahashi, Yanpeng Sun, Kazuya Koyama, Yu Ohtaki, Tetsuhiro Okada, Hidemasa Kawabata, Chiho Maeda, Miyuki Mori, Chiemi Matsumoto, Mayumi Suzuki, Shin-Ichi Chiba, Mishie Tanino, Kenzui Taniue, Hirokazu Kato, Motohiro Yamazaki, Yusuke Mizukami

{"title":"Simultaneous quantitative detection of multiple low-frequency variants by high-dynamic-range capillary electrophoresis.","authors":"Yoshihiko Hagiwara, Nobue Tamamura, Noriyuki Sumida, Yusuke Ono, Kenji Takahashi, Yanpeng Sun, Kazuya Koyama, Yu Ohtaki, Tetsuhiro Okada, Hidemasa Kawabata, Chiho Maeda, Miyuki Mori, Chiemi Matsumoto, Mayumi Suzuki, Shin-Ichi Chiba, Mishie Tanino, Kenzui Taniue, Hirokazu Kato, Motohiro Yamazaki, Yusuke Mizukami","doi":"10.1016/j.omtn.2026.103042","DOIUrl":"10.1016/j.omtn.2026.103042","url":null,"abstract":"Sensitive and quantitative detection of low-frequency variants across multiple loci is critical for nucleic acid-based diagnostics, yet clinical implementation requires a balance among sensitivity, multiplexing capacity, cost, and operational simplicity. We previously developed a high-dynamic-range capillary electrophoresis system capable of detecting variants at allele frequencies below 1%; however, its application was limited to single-locus analysis. Here, we expanded this platform to multiplex detection by incorporating mobility-shift strategies into the assay design. This approach enabled simultaneous analysis of 15 hotspot variants across three clinically relevant loci: KRAS codons 12 and 13 and GNAS codon 201. Validation using synthetic oligonucleotides, formalin-fixed paraffin-embedded tissue, and liquid specimens demonstrated high quantitative accuracy over clinically relevant variant allele frequency ranges, with measured values closely matching expected values (R2 > 0.97). The assay showed high concordance with targeted amplicon sequencing and digital polymerase chain reaction for all variants at variant allele frequencies ≥1%, while also detecting selected variants below this threshold. Collectively, these results establish a multiplexed high-dynamic-range capillary electrophoresis assay for simultaneous, quantitative detection of low-frequency variants, offering a scalable and cost-effective approach for disease-focused gene panels in clinical laboratory settings.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"37 3","pages":"103042"},"PeriodicalIF":6.5,"publicationDate":"2026-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13542593/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148897670","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Harnessing the endosomal escape compartment as a strategy to enhance RNA delivery via lipid nanoparticles. 利用内体逃逸室作为通过脂质纳米颗粒增强RNA递送的策略。

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2026-07-28 eCollection Date: 2026-09-08 DOI: 10.1016/j.omtn.2026.103018

Tom Bourguignon, Preeti Sharma, Inbal Hazan-Halevy, Dan Peer

引用次数: 0

Accurately modeling RNase H-mediated antisense oligonucleotide efficacy. 准确建模RNase h介导的反义寡核苷酸的功效。

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2026-07-07 eCollection Date: 2026-09-08 DOI: 10.1016/j.omtn.2026.103004

Barney Hill, Maisie R Jaques, Remya R Nair, Nicola Whiffin, Matthew J A Wood, Stephan J Sanders, Peter L Oliver, Alyssa C Hill, Carlo Rinaldi

{"title":"Accurately modeling RNase H-mediated antisense oligonucleotide efficacy.","authors":"Barney Hill, Maisie R Jaques, Remya R Nair, Nicola Whiffin, Matthew J A Wood, Stephan J Sanders, Peter L Oliver, Alyssa C Hill, Carlo Rinaldi","doi":"10.1016/j.omtn.2026.103004","DOIUrl":"10.1016/j.omtn.2026.103004","url":null,"abstract":"Antisense oligonucleotides (ASOs) are a powerful class of drugs with the potential to treat a wide range of human diseases. However, the prediction of ASO efficacy remains challenging, as large-scale and costly experimental screens are typically required to identify optimal candidates for a specific therapeutic target. To address this challenge, we compiled ASO Atlas, a database comprising 188,521 RNase H-mediated ASO sequences targeting 334 unique genes with corresponding knockdown efficacy measurements extracted from published patents. Using ASO Atlas, we trained OligoAI, a deep learning model capable of jointly modeling RNA target context, ASO sequence, sugar and backbone chemistries, and dosage to predict in vitro efficacy. We experimentally validated OligoAI by targeting KCNT2, achieving a 5.72-fold reduction in screening effort compared to random selection. ASO Atlas provides the first systematic resource to rigorously evaluate hypotheses regarding key parameters in ASO design, including sequence composition, chemical modifications, and target region selection. Both ASO Atlas and OligoAI have been made freely accessible through an online web-tool with the aim of facilitating the accelerated optimization of ASO design.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"37 3","pages":"103004"},"PeriodicalIF":6.5,"publicationDate":"2026-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13427547/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148653906","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The in vivo CAR-T paradigm shift-Reflections from Boston and the rise of programmable immunity. 体内CAR-T范式的转变——来自波士顿的反思和可编程免疫的兴起。

IF 6.1 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2026-06-09 eCollection Date: 2026-06-16 DOI: 10.1016/j.omtn.2026.102961

Paloma H Giangrande

引用次数: 0

Unlocking translational resilience for mRNA vaccines by viral UTRs. 通过病毒utr解锁mRNA疫苗的翻译弹性。

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2026-06-09 eCollection Date: 2026-06-16 DOI: 10.1016/j.omtn.2026.102969

Jordan J Minnell, Jochem N A Vink, Rebecca E McKenzie

引用次数: 0

Multiscale modeling guided potency assessment of mRNA-lipid nanoparticles. 多尺度建模指导mrna -脂质纳米颗粒效价评估。

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2026-05-26 eCollection Date: 2026-06-16 DOI: 10.1016/j.omtn.2026.102965

Yuling Yang, Yuchen Qiu, Keqi Wang, Yifang Liu, Gautam Sanyal, Paul C Whitford, Sara H Rouhanifard, Wei Xie

{"title":"Multiscale modeling guided potency assessment of mRNA-lipid nanoparticles.","authors":"Yuling Yang, Yuchen Qiu, Keqi Wang, Yifang Liu, Gautam Sanyal, Paul C Whitford, Sara H Rouhanifard, Wei Xie","doi":"10.1016/j.omtn.2026.102965","DOIUrl":"10.1016/j.omtn.2026.102965","url":null,"abstract":"mRNA lipid nanoparticle (mRNA-LNP) technology has emerged as a cornerstone in vaccine development due to its high delivery efficiency, molecular stability, and favorable safety profile. However, rapid and reliable potency assessment remains challenging because of limited mechanistic understanding of delivery processes and sparse experimental data. To address these gaps, we introduce a mechanism-informed, multi-scale kinetic modularized modeling framework that quantitatively captures the coupled dynamics of mRNA delivery across nanoparticle, cellular, and macroscopic scales. The model incorporates variability in LNP-cell interactions and integrates key determinants, such as dosage, LNP and cell size distributions, cell proliferation, and membrane properties-factors that critically shape delivery efficiency and response heterogeneity. Its cell-based architecture and modular design enable adaptability to diverse delivery systems and physiological contexts. By leveraging advanced multi-omics assays, including single-molecule fluorescent in situ hybridization (smFISH) for single-cell resolution of mRNA and protein expression, our framework provides mechanistically grounded modeling and robust prediction of therapeutic potency, offering a powerful platform for optimizing mRNA-based interventions.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"37 2","pages":"102965"},"PeriodicalIF":6.5,"publicationDate":"2026-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13267553/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148265532","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

CD39 mRNA therapy attenuates localized acute inflammation: A novel anti-inflammatory strategy using cationic nanoliposomes. cd39mrna治疗减轻局部急性炎症：一种使用阳离子纳米脂质体的新型抗炎策略。

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2026-05-25 eCollection Date: 2026-06-16 DOI: 10.1016/j.omtn.2026.102966

Naomi R Philosof, Aidan P G Walsh, Bangyan Xu, Anna M D Watson, Shulei Ren, Ludmilla Hann, Benedict Nathaniel, Anne Nguyen, Haikun Liu, Pengkai Shi, Viktoria Bongcaron, Hung Nguyen, Jonathan Noonan, Meltem Avci-Adali, Karlheinz Peter, Mark Louis P Vidallon, Xiaowei Wang

{"title":"CD39 mRNA therapy attenuates localized acute inflammation: A novel anti-inflammatory strategy using cationic nanoliposomes.","authors":"Naomi R Philosof, Aidan P G Walsh, Bangyan Xu, Anna M D Watson, Shulei Ren, Ludmilla Hann, Benedict Nathaniel, Anne Nguyen, Haikun Liu, Pengkai Shi, Viktoria Bongcaron, Hung Nguyen, Jonathan Noonan, Meltem Avci-Adali, Karlheinz Peter, Mark Louis P Vidallon, Xiaowei Wang","doi":"10.1016/j.omtn.2026.102966","DOIUrl":"10.1016/j.omtn.2026.102966","url":null,"abstract":"Messenger RNA (mRNA) therapeutics offer a promising strategy for treating inflammatory disease by enabling transient local expression of therapeutic proteins. CD39 (ectonucleoside triphosphate diphosphohydrolase-1) hydrolyzes pro-inflammatory ATP and ADP and plays a central role in localized immune regulation. We developed cationic nanoliposomes (NLps) for the delivery of CD39 mRNA and evaluated them in a murine model of localized inflammation induced by lipopolysaccharide and matrigel. Biodistribution studies showed substantial retention within the Matrigel matrix, with limited systemic distribution at 24 h. CD39 mRNA-NLps significantly reduced cellular infiltration at day 5, with decreased monocyte and macrophage staining via histological analysis. qPCR confirmed sustained local CD39 mRNA, while flow cytometry demonstrated increased CD39 protein staining in matrigel-derived immune cells, and phosphate release assays showed functional ectonucleotidase activity at the inflammatory site. Interleukin (IL)-6 levels were slightly reduced in matrigel extracts following treatment, supporting suppression of local inflammation. Hemocompatibility was confirmed using ex vivo in human blood and in vivo in treated mice, with liver enzymes and histological analysis supporting formulation's safety and favorable tolerability. These findings establish CD39 mRNA-NLps as a safe, effective strategy for a spatially confined mRNA-based approach to regulate local inflammation.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"37 2","pages":"102966"},"PeriodicalIF":6.5,"publicationDate":"2026-05-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13267555/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148265423","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0