Molecular Therapy. Nucleic Acids最新文献_第6页

FUBP3: A new player in HIV-1 transcriptional activation and immune regulation. FUBP3: HIV-1转录激活和免疫调节的新参与者

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2025-05-27 eCollection Date: 2025-06-10 DOI: 10.1016/j.omtn.2025.102565

Manukumar Honnayakanahalli Marichannegowda, Mohamed S Bouzidi, Satish K Pillai

引用次数: 0

Albumin-binding dendritic siRNA improves delivery and efficacy to solid tumors in a melanoma model. 白蛋白结合树突状siRNA改善了黑色素瘤模型中实体肿瘤的递送和疗效。

IF 6.1 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2025-05-24 eCollection Date: 2025-09-09 DOI: 10.1016/j.omtn.2025.102579

Hassan H Fakih, Qi Tang, Ashley Summers, Katherine Y Gross, Mohamad Omar Rachid, Ken Okamura, Nuria Martinez, Hanadi F Sleiman, John E Harris, Anastasia Khvorova

{"title":"Albumin-binding dendritic siRNA improves delivery and efficacy to solid tumors in a melanoma model.","authors":"Hassan H Fakih, Qi Tang, Ashley Summers, Katherine Y Gross, Mohamad Omar Rachid, Ken Okamura, Nuria Martinez, Hanadi F Sleiman, John E Harris, Anastasia Khvorova","doi":"10.1016/j.omtn.2025.102579","DOIUrl":"10.1016/j.omtn.2025.102579","url":null,"abstract":"Small interfering RNA (siRNA) therapeutics are a new class of drugs that is rapidly expanding to tackle various diseases. Extrahepatic delivery of siRNAs, especially to the parenchyma of solid tumors, is challenging with multiple strategies being explored such as lipid nanoparticle based delivery and ligand conjugation strategies. Here, we report that an albumin-binding dendritic siRNA (D-siRNA) boosts blood circulation time following systemic administration, leading to improved delivery and silencing activity in a melanoma tumor model, in comparison to non-albumin binding lipophilic siRNAs. D-siRNAs increased the tumor-to-liver delivery ratio, including both immune and non-immune cell types within the tumor parenchyma. Using D-siRNAs to target JAK1 expression as an adjuvant to immune checkpoint inhibitors, we found that D-siRNAs was able to enhance PD1 antibody treatment and slow tumor progression of melanoma. Thus, this work demonstrates the utility of D-siRNAs as a systemically administered tumor delivery strategy, enabling the use of siRNAs as chemotherapeutic agents. Further mechanistic studies into the role of JAK1 in melanoma pathology and progression may expand this into additional targets as potential treatments.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"36 3","pages":"102579"},"PeriodicalIF":6.1,"publicationDate":"2025-05-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12213268/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144553977","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

MiR-34 at the crossroads of SMA pathogenesis and therapy: Emerging biomarker and therapeutic target. MiR-34在SMA发病和治疗的十字路口：新兴的生物标志物和治疗靶点。

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2025-05-23 eCollection Date: 2025-06-10 DOI: 10.1016/j.omtn.2025.102557

Jun-An Chen

引用次数: 0

Targeting FANCM using the antisense oligonucleotides to treat the ALT-positive cancers. 利用反义寡核苷酸靶向FANCM治疗alt阳性肿瘤。

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2025-05-23 eCollection Date: 2025-06-10 DOI: 10.1016/j.omtn.2025.102558

Shakir Naji, Christopher Tong, Ashwin Ragupathi, Ming Xiao, Dong Zhang

引用次数: 0

Physicochemical and functional assessment of messenger RNA 5'Cap-end impurities under forced degradation conditions. 在强制降解条件下信使RNA 5' cap端杂质的理化和功能评价。

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2025-05-20 eCollection Date: 2025-06-10 DOI: 10.1016/j.omtn.2025.102570

J Hutchinson, F Schweikart, R Shannon, A Murthy, S M Bates, S Leone, T Nissan, D Sideris, C Lal, J Button, R Rani, G Thom, N Bond, E Örnskov, S Trabulo

{"title":"Physicochemical and functional assessment of messenger RNA 5'Cap-end impurities under forced degradation conditions.","authors":"J Hutchinson, F Schweikart, R Shannon, A Murthy, S M Bates, S Leone, T Nissan, D Sideris, C Lal, J Button, R Rani, G Thom, N Bond, E Örnskov, S Trabulo","doi":"10.1016/j.omtn.2025.102570","DOIUrl":"10.1016/j.omtn.2025.102570","url":null,"abstract":"The 5'Cap is a modified nucleotide structure added to the 5'end of mRNA, which plays a critical role in stability and translation of the molecule. Accurate characterization of 5'Cap impurities is crucial during development of mRNA therapeutics. Here, we utilized orthogonal liquid chromatography-mass spectrometry (LC-MS) workflows to characterize Cap-1-modified mRNA. A complex mixture of uncapped and, notably, 5'Cap chemical degradation variants, was detected. We subjected Cap-1 analogs to process-related forced degradation conditions and characterized multiple degradation routes of the 5'Cap structure, including hydrolysis and depurination. 5'Cap hydrolysis was observed in a representative in vitro transcription (IVT) reaction buffer and was driven by factors including pH, temperature, and the presence of spermidine and MgCl2. Method-induced artifactual degradation occurred during LC-MS data acquisition, emphasizing the importance of using reference standards and optimizing methods to reduce such artifacts. We investigated the effect of 5'Cap degradation on expression and reactogenicity in vitro using model mRNAs with elevated impurity levels. mRNA 5'Cap degradation impurities significantly impacted protein expression but did not trigger innate immune response pathways. 5'Cap degradation impurities should be considered as a critical quality attribute of mRNA therapeutics and should be monitored during production, purification, formulation, and storage.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"36 2","pages":"102570"},"PeriodicalIF":6.5,"publicationDate":"2025-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12166426/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144302540","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

A4GALT-targeting siRNA lipid nanoparticles ameliorate Fabry disease phenotype: Greater efficacy in endothelial cells than in podocytes. 靶向a4galt的siRNA脂质纳米颗粒改善法布里病表型：内皮细胞比足细胞更有效。

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2025-05-20 eCollection Date: 2025-06-10 DOI: 10.1016/j.omtn.2025.102573

Yoo Jin Shin, Hanbi Lee, Xianying Fang, Sheng Cui, Sun Woo Lim, Kang In Lee, Jae Young Lee, Hong Lim Kim, Yuna Oh, Can Li, Chul Woo Yang, Gayeon You, Hyeondo Lee, Hyejung Mok, Byung Ha Chung

{"title":"A4GALT-targeting siRNA lipid nanoparticles ameliorate Fabry disease phenotype: Greater efficacy in endothelial cells than in podocytes.","authors":"Yoo Jin Shin, Hanbi Lee, Xianying Fang, Sheng Cui, Sun Woo Lim, Kang In Lee, Jae Young Lee, Hong Lim Kim, Yuna Oh, Can Li, Chul Woo Yang, Gayeon You, Hyeondo Lee, Hyejung Mok, Byung Ha Chung","doi":"10.1016/j.omtn.2025.102573","DOIUrl":"10.1016/j.omtn.2025.102573","url":null,"abstract":"In this study, we explore the therapeutic feasibility of globotriaosylceramide (Gb3) synthase (A4GALT)-specific siRNA-loaded polyhistidine (pHis)-incorporated lipid nanoparticles (HLNPs) for Fabry disease (FD). HLNPs were developed to deliver siRNAs targeting A4GALT using a microfluidic device, with pHis aiding in endosome escape. The therapy was tested on GLA-knockout human-induced pluripotent-stem-cell-derived endothelial cells (GLA-KO-hiPSC-ECs) and podocytes (GLA-KO-hiPSC-PCs). GLA-KO-hiPSCs-ECs or -PCs, upon differentiation, were treated with A4GALT-siRNA-HLNP. Successful intracellular uptake of A4GALT-siRNA-HLNP was confirmed through fluorescence and electron microscopy in both cell types. A4GALT-siRNA-HLNP treatment confirmed both cell types' stability at 5 μg/mL. Increased Gb3 deposition and zebra body formation were detected in both cell types, but A4GALT-siRNA-HLNP treatment attenuated these FD phenotypes, demonstrating reduced expression of A4GALT through western blot analysis. RNA sequencing analysis revealed that the expression of transcripts associated with FD was restored by A4GALT-siRNA-HLNP treatment in GLA-KO-hiPSCs-ECs, whereas in GLA-KO-hiPSCs-PCs, this effect was relatively less pronounced. Suppression of A4GALT via siRNA/HLNP treatment significantly rescued FD phenotypes especially in EC, presenting a novel therapeutic approach for FD.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"36 2","pages":"102573"},"PeriodicalIF":6.5,"publicationDate":"2025-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12167060/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144302533","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

A novel miniaturized filamentous phagemid as a gene delivery vehicle to target mammalian cells. 一种新型小型化丝状噬菌体作为靶向哺乳动物细胞的基因传递载体。

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2025-05-19 eCollection Date: 2025-06-10 DOI: 10.1016/j.omtn.2025.102571

Shirley Wong, Salma Jimenez, Deborah Pushparajah, Rohini Prakash, Roderick Slavcev

{"title":"A novel miniaturized filamentous phagemid as a gene delivery vehicle to target mammalian cells.","authors":"Shirley Wong, Salma Jimenez, Deborah Pushparajah, Rohini Prakash, Roderick Slavcev","doi":"10.1016/j.omtn.2025.102571","DOIUrl":"10.1016/j.omtn.2025.102571","url":null,"abstract":"The filamentous phage M13 is a single-stranded DNA phage with several attractive characteristics for gene delivery, including a capsid amenable to the display of foreign peptides and a simple well-characterized genome that is easy to genetically modify. Previously, we constructed a DNA minivector based on M13 (a miniphagemid), which minimized the inflammatory bacterial and phage DNA content in the vector. In general, DNA minivectors devoid of their prokaryotic components have shown improved gene transfer and safety. We examined the miniphagemid's capacity for in vitro transgene delivery to target cells through phage display of epidermal growth factor to target its cognate receptor. The absence of the prokaryotic backbone and smaller vector size conferred by the miniphagemids were associated with improved transgene expression for purified single-stranded phagemid DNA and phagemid virion particles. We further engineered this system to enhance packaging of DNA minivectors via deletion of the packaging signal within the helper plasmid used to produce miniphagemids and observed improved phage-mediated gene expression in mammalian cells. Overall, we present a set of novel transgene delivery vectors that combine cell-targeting ligand display and vector minimization. This platform showcases the flexibility of M13 as a gene delivery tool with immense therapeutic potential.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"36 2","pages":"102571"},"PeriodicalIF":6.5,"publicationDate":"2025-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12173652/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144317496","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

Single-stranded HDR templates with truncated Cas12a-binding sequences improve knock-in efficiencies in primary human T cells. 截断cas12a结合序列的单链HDR模板提高了原代人T细胞的敲入效率。

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2025-05-19 eCollection Date: 2025-06-10 DOI: 10.1016/j.omtn.2025.102568

Ana-Maria Nitulescu, Weijie Du, Viktor Glaser, Jonas Kath, Eric J Aird, Grégoire Cullot, Robert Greensmith, Nanna Steengaard Mikkelsen, Maik Stein, Rasmus O Bak, Michael Kaminski, Jacob E Corn, Dimitrios L Wagner

{"title":"Single-stranded HDR templates with truncated Cas12a-binding sequences improve knock-in efficiencies in primary human T cells.","authors":"Ana-Maria Nitulescu, Weijie Du, Viktor Glaser, Jonas Kath, Eric J Aird, Grégoire Cullot, Robert Greensmith, Nanna Steengaard Mikkelsen, Maik Stein, Rasmus O Bak, Michael Kaminski, Jacob E Corn, Dimitrios L Wagner","doi":"10.1016/j.omtn.2025.102568","DOIUrl":"10.1016/j.omtn.2025.102568","url":null,"abstract":"CRISPR-Cas12a gene editing offers an alternative to Cas9-based methods, providing better targeting of AT-rich regions, simplified guide RNA manufacturing, and high specificity. However, the efficacy of donor-based editing is subject to various factors, with template format playing a crucial role. Currently, the predominant non-viral template format for homology-directed repair (HDR) after nuclease-induced DNA breaks is double-stranded DNA, which is toxic when transfected at high doses. Others have demonstrated that using single-stranded DNA (ssDNA) with flanking double-stranded Cas-target-sequences (CTS) as a template for Cas9-mediated gene editing can mitigate this toxicity and increase knock-in efficiency. Here, we investigate CTS design for AsCas12a Ultra by exploring PAM orientation and binding requirements. Additionally, we rule out ssDNase activity of AsCas12a under cell-physiological Mg2+ conditions. Finally, we showcase the advantage of ssDNA donors with CTS (ssCTS) at high doses for delivering clinically relevant transgenes of varying sizes into three TCR-CD3 complex genes (TRAC, CD3ζ, CD3ε), achieving up to 90% knock-in rates for a 0.8kb-insert at the CD3ε locus. Long-read sequencing confirmed higher HDR rates and revealed that CTS reduced partial integration events compared to unmodified ssDNA. Overall, AsCas12a and ssCTS represent a platform for highly efficient knock-in in primary human T cells with minimal toxicity.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"36 2","pages":"102568"},"PeriodicalIF":6.5,"publicationDate":"2025-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12166421/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144302541","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

Targeting CIDEB alleviates liver steatosis and fibrosis in mouse MASH models. 靶向CIDEB可减轻小鼠MASH模型中的肝脏脂肪变性和纤维化。

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2025-05-19 eCollection Date: 2025-06-10 DOI: 10.1016/j.omtn.2025.102567

Yingying Lin, Fushun Fan, Zhenxian Mo, Ziyang Huang, Minhua Zhou, Yaru Ma, Chuiwen Qian, Yifei Wang, Changgeng Qian, Xinjian Liu

{"title":"Targeting CIDEB alleviates liver steatosis and fibrosis in mouse MASH models.","authors":"Yingying Lin, Fushun Fan, Zhenxian Mo, Ziyang Huang, Minhua Zhou, Yaru Ma, Chuiwen Qian, Yifei Wang, Changgeng Qian, Xinjian Liu","doi":"10.1016/j.omtn.2025.102567","DOIUrl":"10.1016/j.omtn.2025.102567","url":null,"abstract":"Cell death-inducing DNA fragmentation factor alpha-like effector B (CIDEB), predominantly expressed in the liver, has been identified as a protective factor against the development of metabolic dysfunction-associated steatohepatitis (MASH) when harboring loss-of-function mutations. In this study, we developed a novel GalNAc-conjugated CIDEB siRNA (GalNAc-siCIDEB) for hepatic delivery to silence CIDEB in the liver, aiming to mimic this protective effect in mouse models. In vitro efficacy screening demonstrated that the siRNA achieved robust silencing of CIDEB across multiple cell lines. In an adeno-associated virus serotype 8 (AAV8)-hCIDEB mouse model, GalNAc-siCIDEB exhibited potent and sustained silencing effect of CIDEB in the hepatocytes, with minimal off-target effects and no observed toxicity. Furthermore, in both high-fat diet-induced obese (HFD-DIO) and choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD)-induced MASH mouse models, GalNAc-siCIDEB effectively reduced lipid droplet formation, suppressed inflammation, and reversed liver steatosis. Additionally, GalNAc-siCIDEB significantly reduced CDAHFD-induced fibrosis in the liver. These findings highlight hepatocyte-specific CIDEB silencing as a promising therapeutic strategy and suggest GalNAc-siCIDEB as a potential candidate for treating MASH.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"36 2","pages":"102567"},"PeriodicalIF":6.5,"publicationDate":"2025-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12156227/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144275428","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

PAI1 regulating CHRNA1 contributes to primary focal hyperhidrosis: Clinical and experimental studies. 临床和实验研究：pa1调节CHRNA1参与原发性局灶性多汗症。

IF 6.5 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2025-05-16 eCollection Date: 2025-06-10 DOI: 10.1016/j.omtn.2025.102566

Ru-Jie Zheng, Nan-Long Lin, Meng-Long Zhang, Rui-Qin Qiu, Feng-Qiang Yu, Xu Li, Jian-Bo Lin

{"title":"PAI1 regulating CHRNA1 contributes to primary focal hyperhidrosis: Clinical and experimental studies.","authors":"Ru-Jie Zheng, Nan-Long Lin, Meng-Long Zhang, Rui-Qin Qiu, Feng-Qiang Yu, Xu Li, Jian-Bo Lin","doi":"10.1016/j.omtn.2025.102566","DOIUrl":"10.1016/j.omtn.2025.102566","url":null,"abstract":"Primary focal hyperhidrosis (PFH) is a debilitating condition characterized by localized excessive sweating, yet its underlying mechanisms remain poorly understood. In this study, sweat gland tissues from PFH patients (n = 204) and healthy controls (n = 60) were analyzed to assess the mRNA and protein levels of plasminogen activator inhibitor 1 (PAI-1) and nicotinic acetylcholine receptor alpha 1 subunit (CHRNA1) using RT-qPCR and western blotting. Primary sweat gland cells were isolated for in vitro experiments, and a pilocarpine-induced hyperhidrosis mouse model was established to evaluate the therapeutic effect of recombinant human PAI-1 (rhPAI-1). PFH patients showed significantly reduced PAI-1 expression and elevated CHRNA1 expression compared to controls (p < 0.01). Treatment with rhPAI-1 downregulated CHRNA1 and aquaporin 5 (AQP5) expression in sweat gland cells and decreased sweat secretion and serum acetylcholine levels in vivo. These results suggest that PAI-1 negatively regulates CHRNA1 and AQP5 expression, offering new insights into the molecular pathology of PFH and identifying PAI-1 as a potential therapeutic target for hyperhidrosis.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"36 2","pages":"102566"},"PeriodicalIF":6.5,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12155562/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144275427","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