Pub Date : 2024-06-26DOI: 10.1016/j.omtn.2024.102263
Christophe Delehedde, Ivan Ciganek, Pierre Louis Bernard, Nabila Laroui, Cathy Costa Da Silva, Cristine Gonçalves, Jacques Nunes, Anne-Lise Bennaceur-Griscelli, Jusuf Imeri, Matthias Huyghe, Luc Even, Patrick Midoux, Nathalie Rameix, Geoffrey Guittard, Chantal Pichon
mRNA applications have undergone unprecedented applications—from vaccination to cell therapy. Natural killer (NK) cells are recognized to have a significant potential in immunotherapy. NK-based cell therapy has drawn attention as allogenic graft with a minimal graft-versus-host risk leading to easier off-the-shelf production. NK cells can be engineered with either viral vectors or electroporation, involving high costs, risks, and toxicity, emphasizing the need for alternative way as mRNA technology. We successfully developed, screened, and optimized novel lipid-based platforms based on imidazole lipids. Formulations are produced by microfluidic mixing and exhibit a size of approximately 100 nm with a polydispersity index of less than 0.2. They are able to transfect NK-92 cells, KHYG-1 cells, and primary NK cells with high efficiency without cytotoxicity, while Lipofectamine Messenger Max and D-Lin-MC3 lipid nanoparticle-based formulations do not. Moreover, the translation of non-modified mRNA was higher and more stable in time compared with a modified one. Remarkably, the delivery of therapeutically relevant interleukin 2 mRNA resulted in extended viability together with preserved activation markers and cytotoxic ability of both NK cell lines and primary NK cells. Altogether, our platforms feature all prerequisites needed for the successful deployment of an NK-based therapeutic strategies.
从疫苗接种到细胞疗法，mRNA 的应用经历了前所未有的发展。自然杀伤（NK）细胞被认为在免疫疗法中具有巨大潜力。基于 NK 的细胞疗法作为异基因移植引起了人们的关注，它的移植物抗宿主风险极低，更易于现成生产。NK 细胞可通过病毒载体或电穿孔技术工程化，但成本高、风险大、毒性强，因此需要采用 mRNA 技术作为替代方法。我们成功开发、筛选并优化了基于咪唑脂质的新型脂质平台。制剂通过微流体混合生产，大小约为 100 纳米，多分散指数小于 0.2。它们能高效转染 NK-92 细胞、KHYG-1 细胞和原代 NK 细胞，且无细胞毒性，而基于 Lipofectamine Messenger Max 和 D-Lin-MC3 脂质纳米颗粒的制剂则不能。此外，与修饰的 mRNA 相比，未修饰的 mRNA 的翻译率更高，时间上也更稳定。值得注意的是，输送治疗相关的白细胞介素 2 mRNA 能延长 NK 细胞系和原代 NK 细胞的存活时间，并保留其活化标志物和细胞毒性能力。总之，我们的平台具备了成功部署基于 NK 的治疗策略所需的所有先决条件。
{"title":"Enhancing natural killer cells proliferation and cytotoxicity using imidazole-based lipid nanoparticles encapsulating interleukin-2 mRNA","authors":"Christophe Delehedde, Ivan Ciganek, Pierre Louis Bernard, Nabila Laroui, Cathy Costa Da Silva, Cristine Gonçalves, Jacques Nunes, Anne-Lise Bennaceur-Griscelli, Jusuf Imeri, Matthias Huyghe, Luc Even, Patrick Midoux, Nathalie Rameix, Geoffrey Guittard, Chantal Pichon","doi":"10.1016/j.omtn.2024.102263","DOIUrl":"https://doi.org/10.1016/j.omtn.2024.102263","url":null,"abstract":"mRNA applications have undergone unprecedented applications—from vaccination to cell therapy. Natural killer (NK) cells are recognized to have a significant potential in immunotherapy. NK-based cell therapy has drawn attention as allogenic graft with a minimal graft-versus-host risk leading to easier off-the-shelf production. NK cells can be engineered with either viral vectors or electroporation, involving high costs, risks, and toxicity, emphasizing the need for alternative way as mRNA technology. We successfully developed, screened, and optimized novel lipid-based platforms based on imidazole lipids. Formulations are produced by microfluidic mixing and exhibit a size of approximately 100 nm with a polydispersity index of less than 0.2. They are able to transfect NK-92 cells, KHYG-1 cells, and primary NK cells with high efficiency without cytotoxicity, while Lipofectamine Messenger Max and D-Lin-MC3 lipid nanoparticle-based formulations do not. Moreover, the translation of non-modified mRNA was higher and more stable in time compared with a modified one. Remarkably, the delivery of therapeutically relevant interleukin 2 mRNA resulted in extended viability together with preserved activation markers and cytotoxic ability of both NK cell lines and primary NK cells. Altogether, our platforms feature all prerequisites needed for the successful deployment of an NK-based therapeutic strategies.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"11 1","pages":""},"PeriodicalIF":8.8,"publicationDate":"2024-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141551492","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}
Pub Date : 2024-06-26eCollection Date: 2024-09-10DOI: 10.1016/j.omtn.2024.102249
Daniel C Nelson, Urmil M Dave, Norberto Gonzalez-Juarbe
{"title":"Harnessing mRNA to unleash endolysins: A new frontier in antibacterial therapy.","authors":"Daniel C Nelson, Urmil M Dave, Norberto Gonzalez-Juarbe","doi":"10.1016/j.omtn.2024.102249","DOIUrl":"10.1016/j.omtn.2024.102249","url":null,"abstract":"","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"35 3","pages":"102249"},"PeriodicalIF":6.5,"publicationDate":"2024-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11260834/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141748650","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}
Ocular neurodegenerative diseases like glaucoma lead to progressive retinal ganglion cell (RGC) loss, causing irreversible vision impairment. Neuroprotection is needed to preserve RGCs across debilitating conditions. Nerve growth factor (NGF) protein therapy shows efficacy, but struggles with limited bioavailability and a short half-life. Here we explore a novel approach to address this deficiency by utilizing circular RNA (circRNA)-based therapy. We show that circRNAs exhibit an exceptional capacity for prolonged protein expression and circRNA-expressed NGF protects cells from glucose deprivation. In a mouse optic nerve crush model, lipid nanoparticle (LNP)-formulated circNGF administered intravitreally protects RGCs and axons from injury-induced degeneration. It also significantly outperforms NGF protein therapy without detectable retinal toxicity. Furthermore, single-cell transcriptomics revealed LNP-circNGF’s multifaceted therapeutic effects, enhancing genes related to visual perception while reducing trauma-associated changes. This study signifies the promise of circRNA-based therapies for treating ocular neurodegenerative diseases and provides an innovative intervention platform for other ocular diseases.
{"title":"Circular RNA-based therapy provides sustained and robust neuroprotection for retinal ganglion cells","authors":"Wenbing Jiang, Dongchang Xiao, Cheng Wu, Jiaqi Yang, Xinghua Peng, Linfeng Chen, Jiamin Zhang, Gaofeng Zha, Wei Li, Rong Ju, Mengqing Xiang, Zhi Xie","doi":"10.1016/j.omtn.2024.102258","DOIUrl":"https://doi.org/10.1016/j.omtn.2024.102258","url":null,"abstract":"Ocular neurodegenerative diseases like glaucoma lead to progressive retinal ganglion cell (RGC) loss, causing irreversible vision impairment. Neuroprotection is needed to preserve RGCs across debilitating conditions. Nerve growth factor (NGF) protein therapy shows efficacy, but struggles with limited bioavailability and a short half-life. Here we explore a novel approach to address this deficiency by utilizing circular RNA (circRNA)-based therapy. We show that circRNAs exhibit an exceptional capacity for prolonged protein expression and circRNA-expressed NGF protects cells from glucose deprivation. In a mouse optic nerve crush model, lipid nanoparticle (LNP)-formulated circNGF administered intravitreally protects RGCs and axons from injury-induced degeneration. It also significantly outperforms NGF protein therapy without detectable retinal toxicity. Furthermore, single-cell transcriptomics revealed LNP-circNGF’s multifaceted therapeutic effects, enhancing genes related to visual perception while reducing trauma-associated changes. This study signifies the promise of circRNA-based therapies for treating ocular neurodegenerative diseases and provides an innovative intervention platform for other ocular diseases.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"49 1","pages":""},"PeriodicalIF":8.8,"publicationDate":"2024-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141551497","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}
Pub Date : 2024-06-17DOI: 10.1016/j.omtn.2024.102259
Mathilde Beaufils, Margaux Melka, Julie Brocard, Clement Benoit, Nagi Debbah, Kamel Mamchaoui, Norma B. Romero, Anne Frédérique Dalmas-Laurent, Susana Quijano-Roy, Julien Fauré, John Rendu, Isabelle Marty
More than 700 pathogenic or probably pathogenic variations have been identified in the gene causing various myopathies collectively known as “-related myopathies.” There is no treatment for these myopathies, and gene therapy stands out as one of the most promising approaches. In the context of a dominant form of central core disease due to a mutation, we aimed at showing the functional benefit of inactivating specifically the mutated allele by guiding CRISPR-Cas9 cleavages onto frequent single-nucleotide polymorphisms (SNPs) segregating on the same chromosome. Whole-genome sequencing was used to pinpoint SNPs localized on the mutant allele and identified specific CRISPR-Cas9 guide RNAs. Lentiviruses encoding these guide RNAs and the nuclease were used to transduce immortalized patient myoblasts, inducing the specific deletion of the mutant allele. The efficiency of the deletion was assessed at DNA and RNA levels, and at the functional level after monitoring calcium release induced by the stimulation of the RyR1-channel. This study provides proof of concept regarding the benefits of mutant allele deletion, in the case of a dominant mutation, from both a molecular and functional perspective, and could apply potentially to 20% of all patients with a mutation.
{"title":"Functional benefit of CRISPR-Cas9-induced allele deletion for RYR1 dominant mutation","authors":"Mathilde Beaufils, Margaux Melka, Julie Brocard, Clement Benoit, Nagi Debbah, Kamel Mamchaoui, Norma B. Romero, Anne Frédérique Dalmas-Laurent, Susana Quijano-Roy, Julien Fauré, John Rendu, Isabelle Marty","doi":"10.1016/j.omtn.2024.102259","DOIUrl":"https://doi.org/10.1016/j.omtn.2024.102259","url":null,"abstract":"More than 700 pathogenic or probably pathogenic variations have been identified in the gene causing various myopathies collectively known as “-related myopathies.” There is no treatment for these myopathies, and gene therapy stands out as one of the most promising approaches. In the context of a dominant form of central core disease due to a mutation, we aimed at showing the functional benefit of inactivating specifically the mutated allele by guiding CRISPR-Cas9 cleavages onto frequent single-nucleotide polymorphisms (SNPs) segregating on the same chromosome. Whole-genome sequencing was used to pinpoint SNPs localized on the mutant allele and identified specific CRISPR-Cas9 guide RNAs. Lentiviruses encoding these guide RNAs and the nuclease were used to transduce immortalized patient myoblasts, inducing the specific deletion of the mutant allele. The efficiency of the deletion was assessed at DNA and RNA levels, and at the functional level after monitoring calcium release induced by the stimulation of the RyR1-channel. This study provides proof of concept regarding the benefits of mutant allele deletion, in the case of a dominant mutation, from both a molecular and functional perspective, and could apply potentially to 20% of all patients with a mutation.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"27 1","pages":""},"PeriodicalIF":8.8,"publicationDate":"2024-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141551521","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}
Pub Date : 2024-06-17DOI: 10.1016/j.omtn.2024.102256
Bogdan Dume, Emilia Licarete, Manuela Banciu
Although recent advancements in cancer immunology have resulted in the approval of numerous immunotherapies, minimal progress has been observed in addressing hard-to-treat cancers. In this context, therapeutic oligonucleotides, including interfering RNAs, antisense oligonucleotides, aptamers, and DNAzymes, have gained a central role in cancer therapeutic approaches due to their capacity to regulate gene expression and protein function with reduced toxicity compared with conventional chemotherapeutics. Nevertheless, systemic administration of naked oligonucleotides faces many extra- and intracellular challenges that can be overcome by using effective delivery systems. Thus, viral and non-viral carriers can improve oligonucleotide stability and intracellular uptake, enhance tumor accumulation, and increase the probability of endosomal escape while minimizing other adverse effects. Therefore, gaining more insight into fundamental mechanisms of actions of various oligonucleotides and the challenges posed by naked oligonucleotide administration, this article provides a comprehensive review of the recent progress on oligonucleotide delivery systems and an overview of completed and ongoing cancer clinical trials that can shape future oncological treatments.
尽管癌症免疫学的最新进展已使许多免疫疗法获得批准，但在治疗难以治疗的癌症方面进展甚微。在这种情况下，治疗性寡核苷酸（包括干扰 RNA、反义寡核苷酸、aptamers 和 DNA 酶）在癌症治疗方法中占据了重要地位，因为与传统化学疗法相比，它们能够调节基因表达和蛋白质功能，而且毒性较低。然而，裸寡核苷酸的全身给药面临着许多细胞外和细胞内的挑战，使用有效的给药系统可以克服这些挑战。因此，病毒和非病毒载体可以改善寡核苷酸的稳定性和细胞内摄取，增强肿瘤积累，提高内体逸出的概率，同时最大限度地减少其他不良反应。因此，为了更深入地了解各种寡核苷酸的基本作用机制以及裸寡核苷酸给药所带来的挑战，本文全面回顾了寡核苷酸给药系统的最新进展，并概述了已完成和正在进行的癌症临床试验，这些试验可能会影响未来的肿瘤治疗。
{"title":"Advancing cancer treatments: The role of oligonucleotide-based therapies in driving progress","authors":"Bogdan Dume, Emilia Licarete, Manuela Banciu","doi":"10.1016/j.omtn.2024.102256","DOIUrl":"https://doi.org/10.1016/j.omtn.2024.102256","url":null,"abstract":"Although recent advancements in cancer immunology have resulted in the approval of numerous immunotherapies, minimal progress has been observed in addressing hard-to-treat cancers. In this context, therapeutic oligonucleotides, including interfering RNAs, antisense oligonucleotides, aptamers, and DNAzymes, have gained a central role in cancer therapeutic approaches due to their capacity to regulate gene expression and protein function with reduced toxicity compared with conventional chemotherapeutics. Nevertheless, systemic administration of naked oligonucleotides faces many extra- and intracellular challenges that can be overcome by using effective delivery systems. Thus, viral and non-viral carriers can improve oligonucleotide stability and intracellular uptake, enhance tumor accumulation, and increase the probability of endosomal escape while minimizing other adverse effects. Therefore, gaining more insight into fundamental mechanisms of actions of various oligonucleotides and the challenges posed by naked oligonucleotide administration, this article provides a comprehensive review of the recent progress on oligonucleotide delivery systems and an overview of completed and ongoing cancer clinical trials that can shape future oncological treatments.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"53 98 1","pages":""},"PeriodicalIF":8.8,"publicationDate":"2024-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141551499","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}
Space particle radiation is a major environmental factor in spaceflight, and it is known to cause body damage and even trigger cancer, but with unknown molecular etiologies. To examine these causes, we developed a systems biology approach by focusing on the co-expression network analysis of transcriptomics profiles obtained from single high-dose (SE) and multiple low-dose (ME) -particle radiation exposures of BEAS-2B human bronchial epithelial cells. First, the differential network and pathway analysis based on the global network and the core modules showed that genes in the ME group had higher enrichment for the extracellular matrix (ECM)-receptor interaction pathway. Then, collagen gene COL1A1 was screened as an important gene in the ME group assessed by network parameters and an expression study of lung adenocarcinoma samples. COL1A1 was found to promote the emergence of the neoplastic characteristics of BEAS-2B cells by both experimental analyses and immunohistochemical staining. These findings suggested that the degree of malignant transformation of cells in the ME group was greater than that of the SE, which may be caused by the dysregulation of the ECM-receptor pathway.
太空粒子辐射是太空飞行中的一个主要环境因素，已知会造成身体损伤，甚至诱发癌症，但分子病因不明。为了研究这些病因，我们开发了一种系统生物学方法，重点对单次高剂量（SE）和多次低剂量（ME）粒子辐射暴露 BEAS-2B 人支气管上皮细胞的转录组学概况进行共表达网络分析。首先，基于全局网络和核心模块的差异网络和通路分析表明，ME 组的基因在细胞外基质（ECM）-受体相互作用通路中的富集度更高。然后，通过网络参数和肺腺癌样本的表达研究，筛选出胶原蛋白基因COL1A1是ME组的重要基因。通过实验分析和免疫组化染色发现，COL1A1能促进BEAS-2B细胞出现肿瘤特征。这些发现表明，ME 组细胞的恶性转化程度高于 SE 组，其原因可能是 ECM 受体通路失调。
{"title":"Differential network analysis reveals the key role of the ECM-receptor pathway in α-particle-induced malignant transformation","authors":"Wenying Yan, Wentao Hu, Yidan Song, Xingyi Liu, Ziyun Zhou, Wanshi Li, Zhifei Cao, Weiwei Pei, Guangming Zhou, Guang Hu","doi":"10.1016/j.omtn.2024.102260","DOIUrl":"https://doi.org/10.1016/j.omtn.2024.102260","url":null,"abstract":"Space particle radiation is a major environmental factor in spaceflight, and it is known to cause body damage and even trigger cancer, but with unknown molecular etiologies. To examine these causes, we developed a systems biology approach by focusing on the co-expression network analysis of transcriptomics profiles obtained from single high-dose (SE) and multiple low-dose (ME) -particle radiation exposures of BEAS-2B human bronchial epithelial cells. First, the differential network and pathway analysis based on the global network and the core modules showed that genes in the ME group had higher enrichment for the extracellular matrix (ECM)-receptor interaction pathway. Then, collagen gene COL1A1 was screened as an important gene in the ME group assessed by network parameters and an expression study of lung adenocarcinoma samples. COL1A1 was found to promote the emergence of the neoplastic characteristics of BEAS-2B cells by both experimental analyses and immunohistochemical staining. These findings suggested that the degree of malignant transformation of cells in the ME group was greater than that of the SE, which may be caused by the dysregulation of the ECM-receptor pathway.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"364 1","pages":""},"PeriodicalIF":8.8,"publicationDate":"2024-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141551500","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}
Successful DNA vaccination generally requires the aid of either a viral vector within vaccine components or an electroporation device into the muscle or skin of the host. However, these systems come with certain obstacles, including limited transgene capacity, broad preexisting immunity in humans, and substantial cell death caused by high voltage pulses, respectively. In this study, we repurposed the use of an amphiphilic bioresorbable copolymer (ABC), called PLA-PEG, as a surface engineering agent that conciliates lipid nanoparticles (LNPs) between stability during preparation and biocompatibility post-vaccination. The LNP carrier can be loaded with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike-specific DNA; in this form, the DNA-LNP is immunogenic in hamsters and elicits protective immunity following DNA-LNP vaccination against heterologous virus challenge or as a hybrid-type vaccine booster against SARS-CoV-2 variants. The data provide comprehensive information on the relationships between LNP composition, manufacturing process, and vaccine efficacy. The outcomes of this study offer new insights into designing next-generation LNP formulations and pave the way for boosting vaccine power to combat existing and possible emerging infectious diseases/pathogens.
{"title":"Boosting DNA vaccine power by lipid nanoparticles surface engineered with amphiphilic bioresorbable copolymer","authors":"Chung-Hsiang Yang, Kuan-Yin Shen, Hui-Min Ho, Chiung-Yi Huang, Yu-Jhen Cheng, Chih-Chun Pu, Fang-Feng Chiu, Wan-Chun Huang, Hung-Chun Liao, Hsin-Wei Chen, Ching-Len Liao, Shih-Jen Liu, Ming-Hsi Huang","doi":"10.1016/j.omtn.2024.102261","DOIUrl":"https://doi.org/10.1016/j.omtn.2024.102261","url":null,"abstract":"Successful DNA vaccination generally requires the aid of either a viral vector within vaccine components or an electroporation device into the muscle or skin of the host. However, these systems come with certain obstacles, including limited transgene capacity, broad preexisting immunity in humans, and substantial cell death caused by high voltage pulses, respectively. In this study, we repurposed the use of an amphiphilic bioresorbable copolymer (ABC), called PLA-PEG, as a surface engineering agent that conciliates lipid nanoparticles (LNPs) between stability during preparation and biocompatibility post-vaccination. The LNP carrier can be loaded with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike-specific DNA; in this form, the DNA-LNP is immunogenic in hamsters and elicits protective immunity following DNA-LNP vaccination against heterologous virus challenge or as a hybrid-type vaccine booster against SARS-CoV-2 variants. The data provide comprehensive information on the relationships between LNP composition, manufacturing process, and vaccine efficacy. The outcomes of this study offer new insights into designing next-generation LNP formulations and pave the way for boosting vaccine power to combat existing and possible emerging infectious diseases/pathogens.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"9 1","pages":""},"PeriodicalIF":8.8,"publicationDate":"2024-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141551496","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}
Pub Date : 2024-06-17DOI: 10.1016/j.omtn.2024.102254
Anna Jonczyk, Marlene Gottschalk, Matthew S.J. Mangan, Yasmin Majlesain, Manja W. Thiem, Lea-Corinna Burbaum, Heike Weighardt, Eicke Latz, Günter Mayer, Irmgard Förster
Allergic contact dermatitis is a prevalent occupational disease with limited therapeutic options. The chemokine CCL22, a ligand of the chemokine receptor CCR4, directs the migration of immune cells. Here, it is shown that genetic deficiency of CCL22 effectively ameliorated allergic reactions in contact hypersensitivity (CHS), a commonly used mouse model of allergic contact dermatitis. For the pharmacological inhibition of CCL22, DNA aptamers specific for murine CCL22 were generated by the systematic evolution of ligands by exponential enrichment (SELEX). Nine CCL22-binding aptamers were initially selected and functionally tested . The 29-nt DNA aptamer AJ102.29m profoundly inhibited CCL22-dependent T cell migration and did not elicit undesired Toll-like receptor-dependent immune activation. AJ102.29m efficiently ameliorated CHS after systemic application. Moreover, CHS-associated allergic symptoms were also reduced following topical application of the aptamer on the skin. Microscopic analysis of skin treated with AJ102.29m demonstrated that the aptamer could penetrate into the epidermis and dermis. The finding that epicutaneous application of the aptamer AJ102.29m in a cream was as effective in suppressing the allergic reaction as intraperitoneal injection paves the way for therapeutic use of aptamers beyond the current routes of systemic administration.
{"title":"Topical application of a CCL22-binding aptamer suppresses contact allergy","authors":"Anna Jonczyk, Marlene Gottschalk, Matthew S.J. Mangan, Yasmin Majlesain, Manja W. Thiem, Lea-Corinna Burbaum, Heike Weighardt, Eicke Latz, Günter Mayer, Irmgard Förster","doi":"10.1016/j.omtn.2024.102254","DOIUrl":"https://doi.org/10.1016/j.omtn.2024.102254","url":null,"abstract":"Allergic contact dermatitis is a prevalent occupational disease with limited therapeutic options. The chemokine CCL22, a ligand of the chemokine receptor CCR4, directs the migration of immune cells. Here, it is shown that genetic deficiency of CCL22 effectively ameliorated allergic reactions in contact hypersensitivity (CHS), a commonly used mouse model of allergic contact dermatitis. For the pharmacological inhibition of CCL22, DNA aptamers specific for murine CCL22 were generated by the systematic evolution of ligands by exponential enrichment (SELEX). Nine CCL22-binding aptamers were initially selected and functionally tested . The 29-nt DNA aptamer AJ102.29m profoundly inhibited CCL22-dependent T cell migration and did not elicit undesired Toll-like receptor-dependent immune activation. AJ102.29m efficiently ameliorated CHS after systemic application. Moreover, CHS-associated allergic symptoms were also reduced following topical application of the aptamer on the skin. Microscopic analysis of skin treated with AJ102.29m demonstrated that the aptamer could penetrate into the epidermis and dermis. The finding that epicutaneous application of the aptamer AJ102.29m in a cream was as effective in suppressing the allergic reaction as intraperitoneal injection paves the way for therapeutic use of aptamers beyond the current routes of systemic administration.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":"157 1","pages":""},"PeriodicalIF":8.8,"publicationDate":"2024-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141507339","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}
Pub Date : 2024-06-17DOI: 10.1016/j.omtn.2024.102257
Ju Hyuen Cha, Seok-Hoon Lee, Yejin Yun, Won Hoon Choi, Hansol Koo, Sung Ho Jung, Ho Byung Chae, Dae Hee Lee, Seok Jae Lee, Dong Hyun Jo, Jeong Hun Kim, Jae-Jin Song, Jong-Hee Chae, Jun Ho Lee, Jiho Park, Jin Young Kang, Sangsu Bae, Sang-Yeon Lee
Mutations in nuclear genes regulating mitochondrial DNA (mtDNA) replication are associated with mtDNA depletion syndromes. Using whole-genome sequencing, we identified a heterozygous mutation (c.272G>A:p.Arg91Gln) in single-stranded DNA-binding protein 1 (SSBP1), a crucial protein involved in mtDNA replisome. The proband manifested symptoms including sensorineural deafness, congenital cataract, optic atrophy, macular dystrophy, and myopathy. This mutation impeded multimer formation and DNA-binding affinity, leading to reduced efficiency of mtDNA replication, altered mitochondria dynamics, and compromised mitochondrial function. To correct this mutation, we tested two adenine base editor (ABE) variants on patient-derived fibroblasts. One variant, NG-Cas9-based ABE8e (NG-ABE8e), showed higher editing efficacy (≤30%) and enhanced mitochondrial replication and function, despite off-target editing frequencies; however, risks from bystander editing were limited due to silent mutations and off-target sites in non-translated regions. The other variant, NG-Cas9-based ABE8eWQ (NG-ABE8eWQ), had a safer therapeutic profile with very few off-target effects, but this came at the cost of lower editing efficacy (≤10% editing). Despite this, NG-ABE8eWQ-edited cells still restored replication and improved mtDNA copy number, which in turn recovery of compromised mitochondrial function. Taken together, base editing-based gene therapies may be a promising treatment for mitochondrial diseases, including those associated with mutations.
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