Molecular Therapy. Nucleic Acids最新文献_第10页

Local administration of a novel siRNA modality into the CNS extends survival and improves motor function in the SOD1G93A mouse model for ALS 在 SOD1G93A ALS 小鼠模型中，向中枢神经系统局部施用新型 siRNA 可延长存活时间并改善运动功能

IF 8.8 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2024-02-15 DOI: 10.1016/j.omtn.2024.102147

Chunling Duan, Moorim Kang, Xiaojie Pan, Zubao Gan, Vera Huang, Guanlin Li, Robert F. Place, Long-Cheng Li

{"title":"Local administration of a novel siRNA modality into the CNS extends survival and improves motor function in the SOD1G93A mouse model for ALS","authors":"Chunling Duan, Moorim Kang, Xiaojie Pan, Zubao Gan, Vera Huang, Guanlin Li, Robert F. Place, Long-Cheng Li","doi":"10.1016/j.omtn.2024.102147","DOIUrl":"https://doi.org/10.1016/j.omtn.2024.102147","url":null,"abstract":"Antisense oligonucleotides (ASOs) were the first modality to pioneer targeted gene knockdown in the treatment of amyotrophic lateral sclerosis (ALS) caused by mutant superoxide dismutase 1 (SOD1). RNA interference (RNAi) is another mechanism of gene silencing in which short interfering RNAs (siRNAs) effectively degrade complementary transcripts. However, delivery to extrahepatic tissues like the central nervous system (CNS) has been a bottleneck in the clinical development of RNAi. Herein, we identify potent siRNA duplexes for the knockdown of human SOD1 (hSOD1) in which medicinal chemistry and conjugation to an accessory oligonucleotide (ACO) enable activity in CNS tissues. Local delivery via intracerebroventricular (ICV) or intrathecal (IT) injection into SOD1 mice delayed disease progression and extended animal survival with superior efficacy compared to an ASO resembling Tofersen in sequence and chemistry. Treatment also prevented disease-related declines in motor function including improvements in animal mobility, muscle strength, and coordination. The ACO itself does not target any specific complementary nucleic acid sequence; rather, it imparts benefits conducive to bioavailability and delivery through its chemistry. The complete conjugate (, siRNA-ACO) represents a novel modality for delivery of duplex RNA (, siRNA) to the CNS that is currently being tested in the clinic for treatment of ALS.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":null,"pages":null},"PeriodicalIF":8.8,"publicationDate":"2024-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139920325","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}

引用次数: 0

MHGTMDA: molecular heterogeneous graph transformer based on biological entity graph for miRNA-disease associations prediction MHGTMDA：基于生物实体图的分子异质图转换器，用于 miRNA 与疾病的关联预测

IF 8.8 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2024-02-05 DOI: 10.1016/j.omtn.2024.102139

Haitao Zou, Boya Ji, Meng Zhang, Fen Liu, Xiaolan Xie, Shaoliang Peng

{"title":"MHGTMDA: molecular heterogeneous graph transformer based on biological entity graph for miRNA-disease associations prediction","authors":"Haitao Zou, Boya Ji, Meng Zhang, Fen Liu, Xiaolan Xie, Shaoliang Peng","doi":"10.1016/j.omtn.2024.102139","DOIUrl":"https://doi.org/10.1016/j.omtn.2024.102139","url":null,"abstract":"MicroRNAs (miRNAs) play a crucial role in the prevention, prognosis, diagnosis, and treatment of complex diseases. Existing computational methods primarily focus on biologically relevant molecules directly associated with miRNA or disease, overlooking the fact that the human body is a highly complex system where miRNA or disease may indirectly correlate with various types of biomolecules. To address this, we propose a novel prediction model named MHGTMDA (miRNA and disease association prediction using Heterogeneous Graph Transformer based on Molecular Heterogeneous Graph). MHGTMDA integrates biological entity relationships of eight biomolecules, constructing a relatively comprehensive heterogeneous biological entity graph. MHGTMDA serves as a powerful molecular heterogeneity map transformer, capturing structural elements and properties of miRNAs and diseases, revealing potential associations. In a 5-fold cross-validation study, MHGTMDA achieved an Area Under the Receiver Operating Characteristic Curve (AUC) of 0.9569, surpassing state-of-the-art methods by at least 3%. Feature ablation experiments suggest that considering features among multiple biomolecules is more effective in uncovering miRNA-disease correlations. Furthermore, we conducted differential expression analyses on breast cancer and lung cancer, using MHGTMDA to further validate differentially expressed miRNAs. The results demonstrate MHGTMDA’s capability to identify novel MDAs. MHGTMDA is free and available at https://github.com/zht-code/HGTMDA.git<svg aria-label=\"Opens in new window\" focusable=\"false\" height=\"8px\" viewbox=\"0 0 8 8\" width=\"8px\"><path d=\"M1.12949 2.1072V1H7V6.85795H5.89111V2.90281L0.784057 8L0 7.21635L5.11902 2.1072H1.12949Z\"></path></svg>.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":null,"pages":null},"PeriodicalIF":8.8,"publicationDate":"2024-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139764639","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}

引用次数: 0

Oligo-PROTAC strategy for cell-selective and targeted degradation of activated STAT3 用于细胞选择性和靶向降解活化 STAT3 的寡聚物-PROTAC 策略

IF 8.8 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2024-02-05 DOI: 10.1016/j.omtn.2024.102137

Jeremy Hall, Zhuoran Zhang, Supriyo Bhattacharya, Dongfang Wang, Marice Alcantara, Yong Liang, Piotr Swiderski, Stephen Forman, Larry Kwak, Nagarajan Vaidehi, Marcin Kortylewski

{"title":"Oligo-PROTAC strategy for cell-selective and targeted degradation of activated STAT3","authors":"Jeremy Hall, Zhuoran Zhang, Supriyo Bhattacharya, Dongfang Wang, Marice Alcantara, Yong Liang, Piotr Swiderski, Stephen Forman, Larry Kwak, Nagarajan Vaidehi, Marcin Kortylewski","doi":"10.1016/j.omtn.2024.102137","DOIUrl":"https://doi.org/10.1016/j.omtn.2024.102137","url":null,"abstract":"Decoy-oligodeoxynucleotides (ODN) allow for targeting undruggable transcription factors, such as STAT3, but their limited potency and lack of delivery methods hampered translation. To overcome these challenges, we conjugated STAT3-specific decoy to thalidomide, a ligand to cereblon in E3 ubiquitin ligase complex, to generate a proteolysis-targeting chimera (STAT3DPROTAC). STAT3DPROTAC downregulated STAT3 in target cells, but not STAT1 or STAT5. Computational modeling of the STAT3DPROTAC ternary complex predicted two surface lysines K601 and K626 in STAT3 as potential ubiquitination sites. Accordingly, K601/K626 point mutations in STAT3, as well as proteasome inhibition or cereblon deletion alleviated STAT3DPROTAC effect. Next, we conjugated STAT3DPROTAC to a CpG oligonucleotide targeting Toll-like receptor 9 (TLR9) to generate myeloid/B-cell-selective C-STAT3DPROTAC. Naked C-STAT3DPROTAC was spontaneously internalized by TLR9+ myeloid cells, B-cells, human and mouse lymphoma cells but not by T cells. C-STAT3DPROTAC effectively decreased STAT3 protein levels and also STAT3-regulated target genes critical for lymphoma cell proliferation and/or survival (BCL2L1, CCND2, MYC). Finally, local C-STAT3DPROTAC administration to human Ly3 lymphoma-bearing mice triggered tumor regression while control C-STAT3D and C-SCR treatments had limited effects. Our results underscore feasibility of using PROTAC strategy for cell-selective, decoy oligonucleotide-based STAT3 targeting of and potentially other tumorigenic transcription factors for cancer therapy.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":null,"pages":null},"PeriodicalIF":8.8,"publicationDate":"2024-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139764750","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}

引用次数: 0

Recent advances in CRISPR/Cas-based genome insertion technologies 基于 CRISPR/Cas 的基因组插入技术的最新进展

IF 8.8 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2024-02-05 DOI: 10.1016/j.omtn.2024.102138

Xinwen Chen, Jingjing Du, Shaowei Yun, Chaoyou Xue, Yao Yao, Shuquan Rao

{"title":"Recent advances in CRISPR/Cas-based genome insertion technologies","authors":"Xinwen Chen, Jingjing Du, Shaowei Yun, Chaoyou Xue, Yao Yao, Shuquan Rao","doi":"10.1016/j.omtn.2024.102138","DOIUrl":"https://doi.org/10.1016/j.omtn.2024.102138","url":null,"abstract":"Programmable genome insertion (or knock-in) is vital for both fundamental and translational research. The continuously expanding number of CRISPR-based genome insertion strategies demonstrates the ongoing development in this field. Common methods for site-specific genome insertion rely on cellular double-strand breaks (DSBs) repair pathways, such as homology directed repair (HDR), non-homologous end joining (NHEJ) and microhomology mediated end joining (MMEJ). Recent advancements have further expanded the toolbox of programmable genome insertion techniques, including prime editing, integrase coupled with programmable nuclease, and CRISPR-associated transposon (CAST). These tools possess their own capabilities and limitations, promoting tremendous efforts to enhance editing efficiency, broaden targeting scope and improve editing specificity. In this review, we first summarize recent advances in programmable genome insertion techniques. We then elaborate on the cons and pros of each technique to assist researchers in making informed choices when utilizing these tools. Finally, we identify opportunities for future improvements and applications in basic research and therapeutics.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":null,"pages":null},"PeriodicalIF":8.8,"publicationDate":"2024-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139764749","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}

引用次数: 0

Anti-miRNA103/107 encapsulated in trasnferrin-conjugated lipid nanoparticles crosses the blood-brain barrier and reduces brain ischemic damage. 包裹在三铁蛋白结合脂质纳米颗粒中的抗miRNA103/107可穿过血脑屏障，减轻脑缺血损伤。

IF 8.8 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2024-02-02 DOI: 10.1016/j.omtn.2024.102131

Pasquale Cepparulo, Ornella Cuomo, Virginia Campani, Antonio Vinciguerra, Maria Josè Sisalli, Valeria Nele, Serenella Anzilotti, Valeria Valsecchi, Antonella Casamassa, Paola Brancaccio, Antonella Scorziello, Giuseppe De Rosa, Lucio Annunziato, Giuseppe Pignataro

{"title":"Anti-miRNA103/107 encapsulated in trasnferrin-conjugated lipid nanoparticles crosses the blood-brain barrier and reduces brain ischemic damage.","authors":"Pasquale Cepparulo, Ornella Cuomo, Virginia Campani, Antonio Vinciguerra, Maria Josè Sisalli, Valeria Nele, Serenella Anzilotti, Valeria Valsecchi, Antonella Casamassa, Paola Brancaccio, Antonella Scorziello, Giuseppe De Rosa, Lucio Annunziato, Giuseppe Pignataro","doi":"10.1016/j.omtn.2024.102131","DOIUrl":"https://doi.org/10.1016/j.omtn.2024.102131","url":null,"abstract":"MicroRNA(miRNA), by post-transcriptionally regulating the expression of genes involved in stroke response, represent important effectors in stroke pathophysiology. Recently, the 103/107 miRNA family emerged as possible therapeutic target in stroke, as it controls the expression of sodium calcium exchanger 1, a plasmamembrane transporter that plays a fundamental role in stroke pathophysiology. Although the neuroprotective properties of this and other miRNAs are promising, several pharmacokinetic drawbacks remain to be faced for the development of a translatable therapy based on small RNAs in CNS diseases. In the present study, to overcome these limitations, the anti-miRNA103/107 was encapsulated in specific preparations of lipid nanoparticles(LNPs), and their effectiveness was evaluated both in an in vitro model of hypoxia represented by primary neuronal cortical cultures exposed to Oxygen and Glucose Deprivation followed by reoxygenation, and in an in vivo model of stroke obtained in rats exposed to transient occlusion of the Middle Cerebral Artery. The results of the present study demonstrated that the encapsulation of anti-miRNA103/107 in transferrin-conjugated PEG-stabilized lipid nanoparticles allowed the blood-brain barrier crossing and significantly reduce brain ischemic damage. The present achievements pave the way for the exploitation of a systemic intravenous miRNA delivery strategy in stroke therapy.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":null,"pages":null},"PeriodicalIF":8.8,"publicationDate":"2024-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139680182","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}

引用次数: 0

Hair Cell-specific Myo15 Promoter-mediated Gene Therapy Rescues Hearing in DFNB9 Mouse Model 毛细胞特异性 Myo15 启动子介导的基因疗法挽救了 DFNB9 小鼠模型的听力

IF 8.8 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2024-02-02 DOI: 10.1016/j.omtn.2024.102135

Hui wang, MengZhao Xun, Honghai Tang, Jingjing Zhao, Shaowei Hu, Longlong Zhang, Jun lv, Daqi Wang, Yuxin Chen, Jianping Liu, Geng-lin Li, Wuqing Wang, Yilai Shu, Huawei Li

{"title":"Hair Cell-specific Myo15 Promoter-mediated Gene Therapy Rescues Hearing in DFNB9 Mouse Model","authors":"Hui wang, MengZhao Xun, Honghai Tang, Jingjing Zhao, Shaowei Hu, Longlong Zhang, Jun lv, Daqi Wang, Yuxin Chen, Jianping Liu, Geng-lin Li, Wuqing Wang, Yilai Shu, Huawei Li","doi":"10.1016/j.omtn.2024.102135","DOIUrl":"https://doi.org/10.1016/j.omtn.2024.102135","url":null,"abstract":"AAV vectors are increasingly used as vehicles for gene delivery to treat hearing loss. However, lack of specificity of the transgene expression may lead to overexpression of the transgene in non-target tissues. In this study, we evaluated the expression efficiency and specificity of transgene delivered by AAV-PHP.eB under the inner ear sensory cell-specific Myo15 promoter. Compared with the ubiquitous CAG promoter, the Myo15 promoter initiates efficient expression of the GFP fluorescence reporter in hair cells, while minimizes non-specific expression in other cell types of the inner ear and central nervous system. Furthermore, using the Myo15 promoter we constructed an AAV-mediated therapeutic system with the coding sequence of OTOF gene. After inner ear injection, we observed apparent hearing recovery in Otof –/– mice, high-efficient expression of exogenous otoferlin, and significant improvement in the exocytosis function of inner hair cells. Overall, our results indicate that gene therapy mediated by the hair cell-specific Myo15 promoter has potential clinical application for the treatment of DFNB9 and yet for other hereditary hearing loss related to dysfunction of hair cells.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":null,"pages":null},"PeriodicalIF":8.8,"publicationDate":"2024-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139670237","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}

引用次数: 0

Tissue pharmacokinetics of antisense oligonucleotides 反义寡核苷酸的组织药代动力学

IF 8.8 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2024-02-02 DOI: 10.1016/j.omtn.2024.102133

Erica Bäckström, Alessandro Bonetti, Per Johnsson, Stefan Öhlin, Anders Dahlén, Patrik Andersson, Shalini Andersson, Peter Gennemark

{"title":"Tissue pharmacokinetics of antisense oligonucleotides","authors":"Erica Bäckström, Alessandro Bonetti, Per Johnsson, Stefan Öhlin, Anders Dahlén, Patrik Andersson, Shalini Andersson, Peter Gennemark","doi":"10.1016/j.omtn.2024.102133","DOIUrl":"https://doi.org/10.1016/j.omtn.2024.102133","url":null,"abstract":"Pharmacokinetics of antisense oligonucleotides (ASO) is characterized by rapid distribution from plasma to tissue and slow terminal plasma elimination driven by re-distribution from tissue. Quantitative understanding of tissue pharmacokinetics and RNA knockdown for various ASO chemistries, conjugations and administration routes is critical for successful drug discovery. Here, we report concentration-time and RNA knockdown profiles for a gapmer ASO with locked nucleic acid ribose chemistry in mouse liver, kidney, heart, and lung after subcutaneous and intratracheal administration. Additionally, the same ASO with liver targeting conjugation (galactosamine-N-acetyl) is evaluated for subcutaneous administration. Data indicate that exposure and knockdown differ between tissues, and strongly depends on administration route and conjugation. In a second study, we show that tissue pharmacokinetics is similar between the three different ribose chemistries locked nucleic acid, constrained ethyl and 2′-O-methoxyethyl, both after subcutaneous and intratracheal administration. Further, we show that the half-life in mouse liver may vary with ASO sequence. Finally, we report less than dose-proportional increase in liver concentration in the dose range 3 to 30 μmol/kg. Overall, our studies contribute pivotal data to support design and interpretation of ASO in vivo studies, thereby increasing the probability of delivering novel ASO therapies to patients.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":null,"pages":null},"PeriodicalIF":8.8,"publicationDate":"2024-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139663761","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}

引用次数: 0

Quantitative proteomics of the miR-301a/SOCS3/STAT3 axis reveals underlying autism and anxiety-like behavior miR-301a/SOCS3/STAT3轴的定量蛋白质组学揭示了自闭症和焦虑样行为的内在原因

IF 8.8 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2024-02-02 DOI: 10.1016/j.omtn.2024.102136

Xun Li, Qi Fu, Mingtian Zhong, Yihao Long, Fengyun Zhao, Yanni Huang, Zizhu Zhang, Min Wen, Kaizhao Chen, Rongqing Chen, Xiaodong Ma

{"title":"Quantitative proteomics of the miR-301a/SOCS3/STAT3 axis reveals underlying autism and anxiety-like behavior","authors":"Xun Li, Qi Fu, Mingtian Zhong, Yihao Long, Fengyun Zhao, Yanni Huang, Zizhu Zhang, Min Wen, Kaizhao Chen, Rongqing Chen, Xiaodong Ma","doi":"10.1016/j.omtn.2024.102136","DOIUrl":"https://doi.org/10.1016/j.omtn.2024.102136","url":null,"abstract":"Autism is a widespread neurodevelopmental disorder. Although the research on ASD has been increasing in the past decade, there is still no specific answer to its mechanism of action and treatment. As a pro-inflammatory microRNA, miR-301a is abnormally expressed in various psychiatric diseases including autism. Here, we show that miR-301a deletion and inhibition exhibited two distinct abnormal behavioral phenotypes in mice. We observed that miR-301a deletion in mice impaired learning/memory, and enhanced anxiety. On the contrary, miR-301a inhibition effectively reduced the maternal immune activation (MIA) induced autism-like behaviors in mice. We further demonstrated that miR-301a bound to the 3’UTR region of the SOCS3, and that inhibition of miR-301a led to the upregulation of SOCS3 in hippocampus. The last result in the reduction of the inflammatory response by inhibiting phosphorylation of AKT and STAT3, and the expression level of IL-17A in poly(I:C)-induced autism-like features in mice. The obtained data revealed the miR-301a as a critical participant in partial behavior phenotypes, which may exhibit a divergent role between gene knockout and knockdown. Our findings ascertain that miR-301a negatively regulate SOCS3 in MIA-induced autism in mice, and could present a new therapeutic target for ameliorating the behavioral abnormalities of autism.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":null,"pages":null},"PeriodicalIF":8.8,"publicationDate":"2024-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139663767","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}

引用次数: 0

Investigating Adverse Genomic and Regulatory Changes Caused by Replacement of the Full-length CFTR cDNA Using Cas9 and AAV 研究使用 Cas9 和 AAV 替换全长 CFTR cDNA 引起的基因组和调控不良变化

IF 8.8 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2024-02-02 DOI: 10.1016/j.omtn.2024.102134

Sriram Vaidyanathan, Jenny L. Kerschner, Alekh Paranjapye, Vrishti Sinha, Brian Lin, Tracy A. Bedrosian, Adrian J. Thrasher, Giandomenico Turchiano, Ann Harris, Matthew H. Porteus

{"title":"Investigating Adverse Genomic and Regulatory Changes Caused by Replacement of the Full-length CFTR cDNA Using Cas9 and AAV","authors":"Sriram Vaidyanathan, Jenny L. Kerschner, Alekh Paranjapye, Vrishti Sinha, Brian Lin, Tracy A. Bedrosian, Adrian J. Thrasher, Giandomenico Turchiano, Ann Harris, Matthew H. Porteus","doi":"10.1016/j.omtn.2024.102134","DOIUrl":"https://doi.org/10.1016/j.omtn.2024.102134","url":null,"abstract":"A \"Universal strategy\" replacing the full-length CFTR cDNA may treat >99% of people with cystic fibrosis (pwCF) regardless of their specific mutations. Cas9-based gene editing was used to insert the CFTR cDNA and a truncated CD19 (tCD19) enrichment tag at the CFTR locus in airway basal stem cells. This strategy restores CFTR function to non-CF levels. Here, we investigate the safety of this approach by assessing genomic and regulatory changes after CFTR cDNA insertion. Safety was first assessed by quantifying genetic rearrangements using CAST-seq. After validating restored CFTR function in edited and enriched airway cells, the CFTR locus open chromatin profile was characterized using ATAC-seq. The regenerative potential and differential gene expression in edited cells was assessed using scRNA-seq. CAST-seq revealed a translocation in ∼0.01% of alleles primarily occurring at a non-oncogenic off-target site and large INDELs in 1% of alleles. The open chromatin profile of differentiated airway epithelial cells showed no appreciable changes except in the region corresponding to the CFTR cDNA and tCD19 cassette indicating no detectable changes in gene regulation. Edited stem cells produced the same types of airway cells as controls with minimal alternations in gene expression. Overall, the Universal strategy showed minor undesirable genomic changes.","PeriodicalId":18821,"journal":{"name":"Molecular Therapy. Nucleic Acids","volume":null,"pages":null},"PeriodicalIF":8.8,"publicationDate":"2024-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139663844","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}

引用次数: 0

mitoTALEN Reduces the Mutant mtDNA Load in Neurons mitoTALEN 可减少神经元中的突变 mtDNA 负荷

IF 8.8 2区医学

Molecular Therapy. Nucleic Acids Pub Date : 2024-02-02 DOI: 10.1016/j.omtn.2024.102132

Sandra R. Bacman, Jose Domingo Barrera-Paez, Milena Pinto, Derek Van Booven, James B. Stewart, Anthony J. Griswold, Carlos T. Moraes

引用次数: 0