Nucleic acid therapeutics最新文献

{"title":"Acknowledgment of Reviewers 2022.","authors":"","doi":"10.1089/nat.2022.29006.ack","DOIUrl":"https://doi.org/10.1089/nat.2022.29006.ack","url":null,"abstract":"","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":"33 1","pages":"81"},"PeriodicalIF":4.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10644006","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Efficient Downregulation of <i>Alk4</i> in Skeletal Muscle After Systemic Treatment with Conjugated siRNAs in a Mouse Model for Duchenne Muscular Dystrophy.","authors":"Sarah Engelbeen,&nbsp;Svetlana Pasteuning-Vuhman,&nbsp;Joke Boertje-van der Meulen,&nbsp;Rubina Parmar,&nbsp;Klaus Charisse,&nbsp;Laura Sepp-Lorenzino,&nbsp;Muthiah Manoharan,&nbsp;Annemieke Aartsma-Rus,&nbsp;Maaike van Putten","doi":"10.1089/nat.2022.0021","DOIUrl":"https://doi.org/10.1089/nat.2022.0021","url":null,"abstract":"<p><p>Downregulation of genes involved in the secondary pathology of Duchenne muscular dystrophy, for example, inflammation, fibrosis, and adiposis, is an interesting approach to ameliorate degeneration of muscle and replacement by fibrotic and adiposis tissue. Small interfering RNAs (siRNAs) are able to downregulate target genes, however, delivery of siRNAs to skeletal muscle still remains a challenge. We investigated delivery of fully chemically modified, cholesterol-conjugated siRNAs targeting <i>Alk4</i>, a nontherapeutic target that is expressed highly in muscle. We observed that a single intravenous or intraperitoneal (IP) injection of 10 mg/kg resulted in significant downregulation of <i>Alk4</i> mRNA expression in skeletal muscles in both wild-type and <i>mdx</i> mice. Treatment with multiple IP injections of 10 mg/kg led to an overall reduction of <i>Alk4</i> expression, reaching significance in tibialis anterior (39.7% ± 6.2%), diaphragm (32.7% ± 5.8%), and liver (41.3% ± 29.9%) in <i>mdx</i> mice. Doubling of the siRNA dose did not further increase mRNA silencing in muscles of <i>mdx</i> mice. The chemically modified conjugated siRNAs used in this study are very promising for delivery to both nondystrophic and dystrophic muscles and could have major implications for treatment of muscular dystrophy pathology.</p>","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":"33 1","pages":"26-34"},"PeriodicalIF":4.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9940804/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10798151","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}

{"title":"Cholesterol-Conjugated siRNA Silencing <i>Tnf</i> for the Treatment of Liver Macrophage-Mediated Acute Inflammation in Nonalcoholic Fatty Liver Disease.","authors":"Kevin Craig,&nbsp;Marc Abrams,&nbsp;Mansoor Amiji","doi":"10.1089/nat.2022.0038","DOIUrl":"https://doi.org/10.1089/nat.2022.0038","url":null,"abstract":"<p><p>Despite wide recognition as a disease of pandemic proportions, effective treatments for nonalcoholic fatty liver disease (NAFLD) remain elusive. Most of the current clinical programs aim to reduce hepatic fat accumulation and, thus, prevent downstream inflammation and fibrosis. To date, this therapeutic approach has helped identify a potential disconnect between steatosis reduction and disease resolution. Mounting preclinical evidence indicates liver inflammation may play a major role in steatosis development and fibrosis but has not garnered the same clinical representation. This may be owing to deficiencies in standard therapeutic modalities that limit their application in NAFLD. RNA interference (RNAi) is an attractive approach to targeting liver inflammation owing to its clinical safety profile, target specificity, and limited biodistribution. In this study, we characterize a simple cholesterol-short-interfering RNA (siRNA) conjugate system targeting <i>Tnf</i> mRNA in liver macrophages for the treatment of NAFLD. First, we observed delivery and anti-inflammatory activity in an acute liver inflammation model. In a follow-up murine NAFLD model, we observed total prevention of nearly all hallmarks of this disease: steatosis, inflammation, and fibrosis. This simple conjugate siRNA delivery system may be the first to show RNAi activity in liver macrophages and provide evidence for a novel therapeutic approach to inflammation in NAFLD.</p>","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":"33 1","pages":"35-44"},"PeriodicalIF":4.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10793808","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Considerations in the Preclinical Assessment of the Safety of Antisense Oligonucleotides.","authors":"Aurélie Goyenvalle,&nbsp;Cecilia Jimenez-Mallebrera,&nbsp;Willeke van Roon,&nbsp;Sabine Sewing,&nbsp;Arthur M Krieg,&nbsp;Virginia Arechavala-Gomeza,&nbsp;Patrik Andersson","doi":"10.1089/nat.2022.0061","DOIUrl":"https://doi.org/10.1089/nat.2022.0061","url":null,"abstract":"<p><p>The nucleic acid therapeutics field has made tremendous progress in the past decades. Continuous advances in chemistry and design have led to many successful clinical applications, eliciting even more interest from researchers including both academic groups and drug development companies. Many preclinical studies in the field focus on improving the delivery of antisense oligonucleotide drugs (ONDs) and/or assessing their efficacy in target tissues, often neglecting the evaluation of toxicity, at least in early phases of development. A series of consensus recommendations regarding regulatory considerations and expectations have been generated by the Oligonucleotide Safety Working Group and the Japanese Research Working Group for the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use S6 and Related Issues (WGS6) in several white papers. However, safety aspects should also be kept in sight in earlier phases while screening and designing OND to avoid subsequent failure in the development phase. Experts and members of the network \"DARTER,\" a COST Action funded by the Cooperation in Science and Technology of the EU, have utilized their collective experience working with OND, as well as their insights into OND-mediated toxicities, to generate a series of consensus recommendations to assess OND toxicity in early stages of preclinical research. In the past few years, several publications have described predictive assays, which can be used to assess OND-mediated toxicity <i>in vitro</i> or <i>ex vivo</i> to filter out potential toxic candidates before moving to <i>in vivo</i> phases of preclinical development, that is, animal toxicity studies. These assays also have the potential to provide translational insight since they allow a safety evaluation in human <i>in vitro</i> systems. Yet, small preliminary <i>in vivo</i> studies should also be considered to complement this early assessment. In this study, we summarize the state of the art and provide guidelines and recommendations on the different tests available for these early stage preclinical assessments.</p>","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":"33 1","pages":"1-16"},"PeriodicalIF":4.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9940817/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10799186","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}

{"title":"Elevation of SHANK3 Levels by Antisense Oligonucleotides Directed Against the 3'-UTR of the Human <i>SHANK3</i> mRNA.","authors":"Nadine Stirmlinger,&nbsp;Jan Philipp Delling,&nbsp;Stefanie Pfänder,&nbsp;Tobias M Boeckers","doi":"10.1089/nat.2022.0048","DOIUrl":"https://doi.org/10.1089/nat.2022.0048","url":null,"abstract":"<p><p>SHANK3 is a member of the SHANK family of scaffolding proteins that localize to the postsynaptic density of excitatory synapses. Mutations within the <i>SHANK3</i> gene or <i>SHANK3</i> haploinsufficiency is thought to be one of the major causes for Phelan-McDermid Syndrome (PMDS) that is characterized by a broad spectrum of autism-related behavioral alterations. Several approaches have already been proposed to elevate SHANK3 protein levels in PMDS patients like transcriptional activation or inhibition of SHANK3 degradation. We undertook a systematic screening approach and tested whether defined antisense oligonucleotides (ASOs) directed against the 3' untranslated region (3'-UTR) of the human <i>SHANK3</i> mRNA are suitable to elevate SHANK3 protein levels. Using human induced pluripotent stem cells (hiPSCs) and hiPSCs-derived motoneurons from controls and PMDS patients we eventually identified two 18 nucleotide ASOs (ASO 4-5.2-4 and 4-5.2-6) that were able to increase SHANK3 protein levels <i>in vitro</i> by about 1.3- to 1.6-fold. These findings were confirmed by co-transfection of the identified ASOs with a GFP-SHANK3-3'-UTR construct in HEK293T cells using GFP protein expression as read-out. Based on these results we propose a novel approach to elevate SHANK3 protein concentrations by 3'-UTR specific ASOs. Further research is needed to test the suitability of <i>SHANK3</i>-specific ASOs as pharmacological compounds also <i>in vivo</i>.</p>","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":"33 1","pages":"58-71"},"PeriodicalIF":4.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9940809/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10793817","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}

{"title":"Consensus Guidelines for the Design and <i>In Vitro</i> Preclinical Efficacy Testing N-of-1 Exon Skipping Antisense Oligonucleotides.","authors":"Annemieke Aartsma-Rus,&nbsp;Alejandro Garanto,&nbsp;Willeke van Roon-Mom,&nbsp;Erin M McConnell,&nbsp;Victoria Suslovitch,&nbsp;Winston X Yan,&nbsp;Jonathan K Watts,&nbsp;Timothy W Yu","doi":"10.1089/nat.2022.0060","DOIUrl":"https://doi.org/10.1089/nat.2022.0060","url":null,"abstract":"<p><p>Antisense oligonucleotides (ASOs) can modulate pre-mRNA splicing. This offers therapeutic opportunities for numerous genetic diseases, often in a mutation-specific and sometimes even individual-specific manner. Developing therapeutic ASOs for as few as even a single patient has been shown feasible with the development of Milasen for an individual with Batten disease. Efforts to develop individualized ASOs for patients with different genetic diseases are ongoing globally. The N = 1 Collaborative (N1C) is an umbrella organization dedicated to supporting the nascent field of individualized medicine. N1C recently organized a workshop to discuss and advance standards for the rigorous design and testing of splice-switching ASOs. In this study, we present guidelines resulting from that meeting and the key recommendations: (1) dissemination of standardized experimental designs, (2) use of standardized reference ASOs, and (3) a commitment to data sharing and exchange.</p>","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":"33 1","pages":"17-25"},"PeriodicalIF":4.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9940807/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9102107","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}

{"title":"Integrated Assessment of Phase 2 Data on GalNAc₃-Conjugated 2'-<i>O</i>-Methoxyethyl-Modified Antisense Oligonucleotides.","authors":"Brenda F Baker,&nbsp;Shuting Xia,&nbsp;Wesley Partridge,&nbsp;T Jesse Kwoh,&nbsp;Sotirios Tsimikas,&nbsp;Sanjay Bhanot,&nbsp;Richard S Geary","doi":"10.1089/nat.2022.0044","DOIUrl":"10.1089/nat.2022.0044","url":null,"abstract":"<p><p>Receptor-mediated delivery of an antisense oligonucleotide (ASO) using the ligand-conjugated antisense technology is establishing a new benchmark for antisense therapeutics. The triantennary <i>N</i>-acetylgalactosamine (GalNAc₃) cluster is the first conjugated ligand to yield a marked increase in ASO potency for RNA targets expressed by hepatocytes, compared to the unconjugated form. In this study, we present an integrated safety assessment of data available from randomized, placebo-controlled, phase 2 studies for six GalNAc₃-conjugated 2'-<i>O</i>-methoxyethyl (2'MOE)-modified ASOs. The total study population included 642 participants (130 placebo; 512 ASO) with up to 1 year of exposure. The primary measures were the incidence of signals from standardized laboratory tests and the mean test results over time. The GalNAc₃-conjugated ASOs were well tolerated with no class effect identified across all doses tested compared to placebo. These results extend prior observations from phase 1 studies, now with treatment up to 1 year.</p>","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":"33 1","pages":"72-80"},"PeriodicalIF":4.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10623620/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10798666","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}

{"title":"Cross-Species Translation of Biophase Half-Life and Potency of GalNAc-Conjugated siRNAs.","authors":"Alessandro Boianelli,&nbsp;Yasunori Aoki,&nbsp;Maxim Ivanov,&nbsp;Anders Dahlén,&nbsp;Peter Gennemark","doi":"10.1089/nat.2022.0010","DOIUrl":"https://doi.org/10.1089/nat.2022.0010","url":null,"abstract":"<p><p>Small interfering RNAs (siRNAs) with <i>N</i>-acetylgalactosamine (GalNAc) conjugation for improved liver uptake represent an emerging class of drugs to treat liver diseases. Understanding how pharmacokinetics and pharmacodynamics translate is pivotal for <i>in vivo</i> study design and human dose prediction. However, the literature is sparse on translational data for this modality, and pharmacokinetics in the liver is seldom measured. To overcome these difficulties, we collected time-course biomarker data for 11 GalNAc-siRNAs in various species and applied the kinetic-pharmacodynamic modeling approach to estimate the biophase (liver) half-life and the potency. Our analysis indicates that the biophase half-life is 0.6-3 weeks in mouse, 1-8 weeks in monkey, and 1.5-14 weeks in human. For individual siRNAs, the biophase half-life is 1-8 times longer in human than in mouse, and generally 1-3 times longer in human than in monkey. The analysis indicates that the siRNAs are more potent in human than in mouse and monkey.</p>","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":"32 6","pages":"507-512"},"PeriodicalIF":4.0,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9784597/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10812713","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}

{"title":"The Quest for mRNA Vaccines.","authors":"Eli Gilboa,&nbsp;David Boczkowski,&nbsp;Smita K Nair","doi":"10.1089/nat.2021.0103","DOIUrl":"https://doi.org/10.1089/nat.2021.0103","url":null,"abstract":"<p><p>The success of mRNA vaccines against COVID-19 is nothing short of a medical revolution. Given its chemical lability the use of mRNA as a therapeutic has been counterintuitive and met with skepticism. The development of mRNA-based COVID-19 vaccines was the culmination of long and painstaking efforts by many investigators spanning over 30 years and culminating with the seminal studies of Kariko and Weissman. This review will describe one chapter in this saga, studies that have shown that mRNA can function as a therapeutic. It started with our seminal observation that dendritic cells (DCs) transfected with mRNA <i>in vitro</i> administered to mice inhibits tumor growth, and led to first-in-human clinical trials with mRNA vaccines in cancer patients. The clinical development of this patient-specific DCs-mRNA approach and use on a larger scale was hindered by the challenges associated with personalized cell therapies. Confirmed and extended by many investigators, these studies did serve as impetus and motivation that led scientists to persevere, eventually leading to the development of simple, broadly applicable, and highly effective protocols of directly injecting mRNA into patients, culminating in the COVID-19 mRNA vaccines.</p>","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":"32 6","pages":"449-456"},"PeriodicalIF":4.0,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10419344","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Early-Stage Identification and Avoidance of Antisense Oligonucleotides Causing Species-Specific Inflammatory Responses in Human Volunteer Peripheral Blood Mononuclear Cells.","authors":"Sebastien A Burel,&nbsp;Todd Machemer,&nbsp;Brenda F Baker,&nbsp;T Jesse Kwoh,&nbsp;Suzanne Paz,&nbsp;Husam Younis,&nbsp;Scott P Henry","doi":"10.1089/nat.2022.0033","DOIUrl":"https://doi.org/10.1089/nat.2022.0033","url":null,"abstract":"<p><p>A human peripheral blood mononuclear cell (PBMC)-based assay was developed to identify antisense oligonucleotide (ASO) with the potential to activate a cellular innate immune response outside of an acceptable level. The development of this assay was initiated when ISIS 353512 targeting the messenger ribonucleic acid for human C-reactive protein (CRP) was tested in a phase I clinical trial, in which healthy human volunteers unexpectedly experienced increases in interleukin-6 (IL-6) and CRP. This level of immune stimulation was not anticipated following rodent and nonhuman primate safety studies in which no evidence of exaggerated proinflammatory effects were observed. The IL-6 increase induced by ISIS 353512 was caused by activation of B cells. The IL-6 induction was inhibited by chloroquine pretreatment of PBMCs and the nature of ASOs suggested that the response is mediated by a Toll-like receptor (TLR), in all likelihood TLR9. While assessing the inter PBMC donor variability, two classes of human PBMC responders to ISIS 353512 were identified (discriminator and nondiscriminators). The discriminator donor PBMCs were shown to produce low level of IL-6 after 24 h in culture, in the absence of ASO treatment. The PBMC assay using discriminator donors was shown to be reproducible, allowing to assess reliably the immune potential of ASOs by comparison to known benchmark ASO controls that were previously shown to be either safe or inflammatory in clinical trials. Clinical Trial registration numbers: NCT00048321 NCT00330330 NCT00519727.</p>","PeriodicalId":19412,"journal":{"name":"Nucleic acid therapeutics","volume":"32 6","pages":"457-472"},"PeriodicalIF":4.0,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10432044","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}