作者: Li, Anqi ; Pan, Qingshan ; Zhang, Yan ; Zhang, Xuan ; Qiao, Dan ; Xu, Shan ; Peng, Rujue ; Wan, Li ; Zheng, Pengwu ; Li, Na ; Zhu, Wufu ; Guo, Qiuyan

Bacterial infection poses a significant challenge to wound healing and skin regeneration, leading to substantial economic burdens on patients and society. Therefore, it is crucial to promptly explore and develop effective methodologies for bacterial infections. Herein, we propose a novel approach for synthesizing nanostructures based on antisense oligonucleotides (ASOs) through the coordination-driven self-assembly of Zn²⁺ with ASO molecules. This approach aims to provide effective synergistic therapy for chronic wound infections caused by Staphylococcus aureus (S. aureus). The resulting hybrid nanoparticles successfully preserve the structural integrity and biological functionalities of ASOs, demonstrating excellent ASO encapsulation efficiency and bioaccessibility. In vitro antibacterial experiments reveal that Zn-ASO NPs exhibit antimicrobial properties against Escherichia coli, Staphylococcus aureus, and Bacillus subtilis. This antibacterial ability is attributed to the high concentration of metal zinc ions and the generation of high levels of reactive oxygen species. Additionally, the ftsZ-ASO effectively inhibits the expression of the ftsZ gene, further enhancing the antimicrobial effect. In vivo antibacterial assays demonstrate that the Zn-ASO NPs promote optimal skin wound healing and exhibit favorable biocompatibility against S. aureus infections, resulting in a residual infected area of less than 8%. This combined antibacterial strategy, which integrates antisense gene therapy and metal-coordination-directed self-assembly, not only achieves synergistic and augmented antibacterial outcomes but also expands the horizons of ASO coordination chemistry. Moreover, it addresses the gap in the antimicrobial application of metal-coordination ASO self-assembly, thereby advancing the field of ASO-based therapeutic approaches.

2024-03-01·Talanta

Theranostic DNA nanostructure based on phenotype-specific activation of antisense oligonucleotides

Article

作者: Chu, Lok Ting ; Chen, Ting-Hsuan ; Kwong, Hoi Kwan ; Cui, Chenyu

Antisense oligonucleotide (ASO) is a powerful agent for gene therapy, designed to form complementary pairs with specific mRNA to inhibit gene expression. However, low specificity limits its potential. To overcome this challenge, we developed a Y-shape DNA nanostructure that enhances the specificity in ASO-based treatment by introducing a detection trigger. The design incorporates the phenotype-specific miR21 activation and the sequential release of Bcl2 ASO. As a result, our Y-shape DNA nanostructure downregulates >50 % Bcl2 mRNA expression and induces >60 % cell death in breast cancer cells. Meanwhile, this approach shows no obvious damage to the non-cancerous cells, indicating the therapeutic potential as a theranostics agent in precision medicine with the combination of biomarker sensing and treatment. Overall, our Y-shape DNA nanostructure serves as a promising strategy providing potential in customized conformation design with specific target sequences in gene therapy.

2023-01-01·Revue Neurologique

Therapeutic tools for familial ALS

Review

作者: Camu, W ; Esselin, F ; De La Cruz, E

Familial ALS (FALS) accounts for 10 to 15% of ALS cases. In more than 70% of FALS patients, a causal gene is identified and animal models have been developed for a subset of them, mainly for the most frequently mutated genes. Therapeutic tools to treat those patients are dominated by gene-specific therapy and the most advanced approaches target the SOD1 gene mutations. Either by direct delivery of antisense oligonucleotides (ASO) or using viral vectors such as adenoviruses (AAV) to deliver ASOs, gene specific therapies have shown promising results in animal models. The recent use of subpial injections of AAV9+anti SOD1 ASO now shows that the disease is completely prevented or stopped in the animal, depending on the moment of injection, e.g., before or after disease onset. However, the use of viral vectors in humans seems to be limited at least by their immunogenicity. Antibody-based therapies are also efficient to treat animal models, but to a lesser extent. Most of the experiments targeted the SOD1 protein in its misfolded conformation. This approach seems better tolerated than the AAV one, an important limit being the choice of the epitope. Unexpectedly, some advances in treating the C9ORF72 animal model have been obtained using a modulation of microbiota, and this strategy has the great advantage to have an easy route of administration and a good safety profile. The landscape of experimental FALS treatment is rapidly evolving and results are promising. This is an important unmet need for ALS patients and several human phase I, II and III trials are ongoing.

项与 ALG ASO 相关的新闻（医药）

2023-05-03

·EndPoints

ARTHEx nabs Series B for clinical trial of ASO in myotonic dystrophy as Vertex’s deals heat up the space

ARTHEx Biotech has raised €42 million (about $46 million) in Series B funds to bankroll an early-stage study of a drug candidate in myotonic dystrophy type 1, or DM1. The Valencia, Spain-based startup will enter the clinic in the second half of this year with an IV-delivered drug candidate for people with the rare neuromuscular disorder. Muscle weakness is a common result of DM1 and impacts about 40,000 people in the US and 70,000 in Europe. Those are the two geographies where ARTHEx will investigate ATX-01 in Phase I/IIa, said chair and new CEO Frédéric Legros in an interview. Unlock this story instantly and join 167,400+ biopharma pros reading Endpoints daily — and it's free.

2023-04-07

·生物谷

EMBO Mol Med：增强大脑中冷休克蛋白水平或有望治疗阿尔兹海默症等一系列人类神经退行性疾病

如今，神经退行性疾病在老年人群中越来越普遍，但目前尚无相应的治疗方法；近日，一篇发表在国际杂志EMBO Molecular Medicine上题为“ASO targeting RBM3 temperature‐controlled poison exon splicing prevents neurodegeneration in vivo”的研究报告中，来自柏林自由大学等机构的科学家们通过研究利用一种新型的基因疗法成功增加了小鼠大脑中所谓的“冷休克蛋白”（cold shock protein）的水平，并能保护其抵御朊病毒病所带来的潜在破坏性影响。这一研究发现或许是研究人员利用冷却大脑的保护性效应来治疗急性大脑损伤甚至是预防诸如阿尔兹海默病等人类痴呆症迈出的重要一步。当机体大幅降温时就会增加名为RBM3的冷休克蛋白的水平，这种现象首次是在冬眠动物中观察到的；科学家们认为，在冬眠期间，这种蛋白能帮助保护大脑免于损伤并促使其继续形成新的神经元连接。早在2015年，研究者Giovanna Mallucci教授等人就通过对小鼠进行研究后发现，RBM3能保护大脑抵御与错误折叠蛋白积累相关的损伤，蛋白质的错误折叠或许会导致多种形式痴呆症的发生，比如阿尔兹海默病和帕金森疾病以及诸如克雅二氏症（CJD）等朊病毒病。诱导性低温用于在重症监护室对患者进行治疗，包括新生儿和创伤性脑损伤患者，即让病人处于昏迷状态，这样其大脑就能被冷却从而防止损伤发生，但也会伴随出现其它风险，比如血液凝固和肺炎等，那么是否能利用冷休克蛋白来治疗病人而不必冷却机体，从而就能为急性脑损伤患者提供更为安全的疗法，或能保护大脑抵御痴呆症呢？这项研究中，研究人员研究了名为反义寡核苷酸(ASOs，antisense oligonucleotides)的基因疗法形式是否能增加小鼠大脑中冷休克蛋白的水平从而保护其大脑免于损伤。研究人员分析了能编码冷休克蛋白产生的特殊基因，结果发现，其包含了一种在正常情况下能阻断其表达的关键元素，利用ASO移除或调低该元件就会使得RBM3的产生得到持久性的提升。为了测试是否这种方法能保护大脑免于损伤，研究人员利用朊病毒感染的小鼠进行研究，其中一些小鼠在3周后被注射了单一剂量的ASO，而其它小鼠则接受了对照疗法进行治疗。在注射朊病毒12周后，接受对照疗法的小鼠患上了朊病毒病，且表现出了海马体中神经元的广泛缺失，海马体是大脑中负责记忆的关键区域。而接受ASO治疗的小鼠的情况则非常不同，当其它小鼠屈服于朊病毒病的同时，接受ASO治疗的小鼠机体中RBM3的水平是其它小鼠的两倍，8只接受ASO治疗的小鼠中有7只小鼠都表现为海马体中神经元的广泛保护。通过增强机体大脑中冷休克蛋白的水平或有望治疗一系列人类神经退行性疾病。图片来源：EMBO Molecular Medicine (2023). DOI:10.15252/emmm.202217157研究者Giovanna Mallucci教授说道，从本质上来讲，冷休克蛋白能促使大脑自我保护，在这种情况下就能抵御朊病毒病期间对大脑中神经细胞的损伤；而值得注意的是，本文研究结果表明，仅仅单一注射ASO或许就足以为这些小鼠提供长效的保护力，并预防不可避免的神经退行性疾病的发生和发展。这种方法或许能为保护机体抵御诸如阿尔兹海默病和帕金森疾病等神经退行性疾病提供新的希望，对于这些疾病而言，目前研究人员并没有可靠的预防性疗法。由于本文研究是在小鼠机体中进行的，所以距离在人类中进行相关的试验还需要很长一段路要走，但如果研究人员能安全利用ASOs来增强人类机体中冷休克蛋白的产生，或许就有望帮助预防人类痴呆症的发生；如今研究人员也看到了ASOs的确能成功用于治疗脊髓性肌肉萎缩症（spinal muscular atrophy），而且其最近还被授权用于治疗人类的运动神经元疾病。如果这些研究发现能在人类中得到复制的话，这一方法或许对于治疗人类神经退行性疾病以外的其它疾病也具有重大意义，包括在心脏手术中通过保护大脑而引起的新生儿缺氧所致急性脑损伤、中风以及成年人的头部损伤，否则将会通过治疗性低温来对这些病症进行治疗。综上，本文在小鼠研究中所得到的可喜成果或许支持了这样一种可能性，即RBM3所诱导的ASO或许可能在从急性脑损伤到阿尔兹海默病等一系列疾病患者机体中提供一定的神经保护作用。原始出处：Marco Preußner，Heather L Smith，Daniel Hughes, et al. ASO targeting RBM3 temperature‐controlled poison exon splicing prevents neurodegeneration in vivo, EMBO Molecular Medicine (2023). DOI: 10.15252/emmm.202217157

基因疗法寡核苷酸

2023-02-20

·生物谷

警惕!脑脊液途径反义寡核苷酸会引起急性神经毒性

反义寡核苷酸(ASO)是人工合成的单链DNA或RNA序列，可以与靶RNA以Watson-Crick的方式杂交，从而改变RNA的加工过程以调节蛋白质的产生。反义寡核苷酸(ASO)是人工合成的单链DNA或RNA序列，可以与靶RNA以Watson-Crick的方式杂交，从而改变RNA的加工过程以调节蛋白质的产生。由于ASO的电荷和大小使其很难通过血脑屏障，因此已开发出通过鞘内注射将ASO直接注入脑脊液(CSF)间隙来治疗中枢神经系统(CNS)疾病的方法。由于ASO的电荷和大小使其很难通过血脑屏障，因此已开发出通过鞘内注射将ASO直接注入脑脊液(CSF)间隙来治疗中枢神经系统(CNS)疾病的方法。反义寡核苷酸(ASO)是治疗中枢神经系统(CNS)顽固性疾病的有效药物。对于这种临床应用，神经毒性是一个关键的限制因素。因此，从行为学的角度评估这种神经毒性对于揭示中枢神经系统的症状性功能障碍和阐明其潜在的分子机制具有重要意义。图片来源: https://doi-org.libproxy1.nus.edu.sg/10.1016/j.omtn.2022.12.010 近日，来自东京医科大学的研究者们在Molecular Therapy Nucleic Acids杂志上发表了题为“Change of intracellular calcium level causes acute neurotoxicity by antisense oligonucleotides via CSF route”的文章，该研究结果证实了ASO神经毒性的行为学特征，并揭示了细胞内游离钙水平的变化是ASO神经毒性作用的机制之一。反义寡核苷酸(ASO)是治疗中枢神经系统(CNS)顽固性疾病的有效药物。对于这种临床应用，神经毒性是一个关键的限制因素。因此，从行为学的角度评估这种神经毒性对于揭示中枢神经系统的症状性功能障碍和阐明其潜在的分子机制具有重要意义。在本研究中，研究者利用行为分析方法对通过脑室注射有毒ASO的小鼠的急性神经毒性进行分类和量化。毒性ASO以剂量依赖的方式降低小鼠的意识和运动功能。研究者从机制上分析了调节神经元钙内流的受体或通道调节剂对ASO神经毒性的影响。促进钙内流的调节剂减轻了ASOS的体内神经毒性，而阻碍钙内流的调节剂则增加了。在一项利用大鼠原代皮质神经元评估细胞内游离钙水平的体外实验中，有毒的ASO降低了钙水平。静脉注射ASOS后可引起低活性神经毒性图片来源: https://doi-org.libproxy1.nus.edu.sg/10.1016/j.omtn.2022.12.010 本研究已经建立了一种行为评估方法，用来表征I.C.V。给予ASO诱导的中枢神经系统毒性，研究者揭示了其潜在的机制之一，即钙离子的介导。研究者认为额外给予钙通道激动剂或AMPAR激动剂可以在不降低靶基因沉默效果的情况下减轻ASO的神经毒性，这是一种有希望的策略，可以在体内提高ASOS针对中枢神经系统的治疗指数。由于ASOS的中枢神经系统毒性是其临床应用的关键限制之一，本研究结果可能为克服这一局限性提供新的见解，扩大治疗窗口，并将导致开发ASOS靶向中枢神经系统疾病的新策略和新技术。（生物谷 Bioon.com）参考文献 Chunyan Jia et al. Change of intracellular calcium level causes acute neurotoxicity by antisense oligonucleotides via CSF route. Mol Ther Nucleic Acids. 2022 Dec 23;31:182-196. doi: 10.1016/j.omtn.2022.12.010.